This document summarizes a study that evaluated five different techniques for processing umbilical cord blood stem cells: plasma depletion, density gradient separation, Hetastarch separation, a novel PrepaCyte-CB method, and an automated Sepax system. The study found that Sepax achieved the highest recovery of nucleated cells at 78.8%, while PrepaCyte-CB achieved the highest recovery of CD34+ hematopoietic stem cells at 74.47%. Density gradient separation was the most effective at volume reduction, while PrepaCyte-CB processing left samples with the highest clonogenic potential after processing and cryopreservation. The study concluded that the choice of processing method is important and depends on the
Machine pulsatile perfusion is a novel technique for isolating stem cells from the entire afterbirth, including placenta and umbilical cord, which results in a 2-5x higher yield of CD34+ stem cells compared to cord blood alone. The technique mimics the human heart and gently extracts stem cells using perfusate chemistry matched to normal body conditions. Studies show afterbirth-derived stem cells isolated using this method have similar performance to cord blood stem cells for blood and immune cell production and it is being explored as a new source of stem cells for transplantation therapies.
In-vitro Cell culture is developing rapidly with the primary goal being to model human physiology in a realistic way that predicts clinical results. This presentation shares the strategy being employed by Kirkstall Ltd to provide a practical solution to the challenges of creating an organ on-a-chip.
The umbilical cord matrix is a better source of mesenchymal stem cells than umbilical cord blood based on the following:
1) Mesenchymal stem cells were successfully isolated from 8 out of 8 umbilical cord matrix samples but only 1 out of 15 umbilical cord blood samples when using various isolation and culture methods.
2) Cells isolated from umbilical cord blood consisted mainly of macrophage-like cells and spindle-shaped cells that lacked mesenchymal stem cell markers, whereas cells from umbilical cord matrix and bone marrow formed fibroblast colonies with mesenchymal stem cell characteristics.
3) The expansion potential of mesenchymal stem cells was highest
WEBINAR Characterisation of human pluripotent stem cells (ESCs and IPSC) and ...Quality Assistance s.a.
Valérie DEFFONTAINE, R&D Scientist, Quality Assistance
Webinar held on 8th June 2017.
The discovery of human pluripotent stem cells 10 years ago turned the spotlight on the potential of pluripotent stem cells for personalised cell therapy. The scientific interest then quickly shifted towards the use of these cells for safety pharmacology, drug discovery and disease modelling. For all these purposes, in the mid to long term, properly characterised cell banks will be necessary.
The characterisation of embryonic (ESC) and induced pluripotent stem cells (IPSC) used for manufacturing requires the development and validation of analytical methods (e.g. flow cytometry, microscopy, QPCR and bioassays). Cell characterisation includes the testing of cell product identity, determination of impurities, and assessment of biological activity and viability. Among the techniques available, flow cytometry is widely used to assess the expression of cell markers. Our laboratory has developed flow cytometry panels dedicated to the characterisation of extracellular and intracellular markers of ESC and IPSC, and to the detection of cell-related impurities. We proposed a method for the validation of flow cytometry panels according to the recommendations of international guidelines on the validation of analytical methods.
IPSC differentiated into cardiomyocytes and MSC-like cells were also used to test the performance of our flow cytometry panels to accurately monitor the manufacturing process of cell products.
In addition to the technical tips, this webinar aims at presenting a critical view on the use of flow cytometry platform for cell characterisation.
For more information, visit http://www.quality-assistance.com/analytical-services/CBMPs
14 rencontres biomédicale LIR Philippe MenaschéAssociation LIR
The document discusses stem cell research in France for cardiovascular diseases. It outlines France's long experience in cell therapy and efforts to streamline clinical trials. It describes research on embryonic stem cells and cell transplantation trials for heart failure. It emphasizes lessons from early experiences, such as tailoring cell choice to indications and optimizing cell engraftment. Finally, it discusses opportunities for induced pluripotent stem cell research based on existing ESC expertise in France.
The collection and processing of hematopoieticakshaya tomar
BASICS OF HSC COLLECTION AND PROCESSING INCLUDING ALL THE THREE SOURCES, A BRIEF ABOUT STEM CELL MOBILIZATION, STEM CELL SELECTION CRYOPRESERVATION AND DMSO
This document summarizes the connections between clinical trials and family banking of perinatal stem cells. It reviews the first decade of advanced cell therapy clinical trials using perinatal stem cells from 2005-2015. It also surveys the perinatal cell storage services offered by family cord blood banks, including what cell and tissue types they store. The growth of family cord blood banking has helped drive clinical trials by providing a source of autologous cells and funding from bank owners seeking to justify the family banking market.
Machine pulsatile perfusion is a novel technique for isolating stem cells from the entire afterbirth, including placenta and umbilical cord, which results in a 2-5x higher yield of CD34+ stem cells compared to cord blood alone. The technique mimics the human heart and gently extracts stem cells using perfusate chemistry matched to normal body conditions. Studies show afterbirth-derived stem cells isolated using this method have similar performance to cord blood stem cells for blood and immune cell production and it is being explored as a new source of stem cells for transplantation therapies.
In-vitro Cell culture is developing rapidly with the primary goal being to model human physiology in a realistic way that predicts clinical results. This presentation shares the strategy being employed by Kirkstall Ltd to provide a practical solution to the challenges of creating an organ on-a-chip.
The umbilical cord matrix is a better source of mesenchymal stem cells than umbilical cord blood based on the following:
1) Mesenchymal stem cells were successfully isolated from 8 out of 8 umbilical cord matrix samples but only 1 out of 15 umbilical cord blood samples when using various isolation and culture methods.
2) Cells isolated from umbilical cord blood consisted mainly of macrophage-like cells and spindle-shaped cells that lacked mesenchymal stem cell markers, whereas cells from umbilical cord matrix and bone marrow formed fibroblast colonies with mesenchymal stem cell characteristics.
3) The expansion potential of mesenchymal stem cells was highest
WEBINAR Characterisation of human pluripotent stem cells (ESCs and IPSC) and ...Quality Assistance s.a.
Valérie DEFFONTAINE, R&D Scientist, Quality Assistance
Webinar held on 8th June 2017.
The discovery of human pluripotent stem cells 10 years ago turned the spotlight on the potential of pluripotent stem cells for personalised cell therapy. The scientific interest then quickly shifted towards the use of these cells for safety pharmacology, drug discovery and disease modelling. For all these purposes, in the mid to long term, properly characterised cell banks will be necessary.
The characterisation of embryonic (ESC) and induced pluripotent stem cells (IPSC) used for manufacturing requires the development and validation of analytical methods (e.g. flow cytometry, microscopy, QPCR and bioassays). Cell characterisation includes the testing of cell product identity, determination of impurities, and assessment of biological activity and viability. Among the techniques available, flow cytometry is widely used to assess the expression of cell markers. Our laboratory has developed flow cytometry panels dedicated to the characterisation of extracellular and intracellular markers of ESC and IPSC, and to the detection of cell-related impurities. We proposed a method for the validation of flow cytometry panels according to the recommendations of international guidelines on the validation of analytical methods.
IPSC differentiated into cardiomyocytes and MSC-like cells were also used to test the performance of our flow cytometry panels to accurately monitor the manufacturing process of cell products.
In addition to the technical tips, this webinar aims at presenting a critical view on the use of flow cytometry platform for cell characterisation.
For more information, visit http://www.quality-assistance.com/analytical-services/CBMPs
14 rencontres biomédicale LIR Philippe MenaschéAssociation LIR
The document discusses stem cell research in France for cardiovascular diseases. It outlines France's long experience in cell therapy and efforts to streamline clinical trials. It describes research on embryonic stem cells and cell transplantation trials for heart failure. It emphasizes lessons from early experiences, such as tailoring cell choice to indications and optimizing cell engraftment. Finally, it discusses opportunities for induced pluripotent stem cell research based on existing ESC expertise in France.
The collection and processing of hematopoieticakshaya tomar
BASICS OF HSC COLLECTION AND PROCESSING INCLUDING ALL THE THREE SOURCES, A BRIEF ABOUT STEM CELL MOBILIZATION, STEM CELL SELECTION CRYOPRESERVATION AND DMSO
This document summarizes the connections between clinical trials and family banking of perinatal stem cells. It reviews the first decade of advanced cell therapy clinical trials using perinatal stem cells from 2005-2015. It also surveys the perinatal cell storage services offered by family cord blood banks, including what cell and tissue types they store. The growth of family cord blood banking has helped drive clinical trials by providing a source of autologous cells and funding from bank owners seeking to justify the family banking market.
This literature review finds that umbilical cord tissue, specifically Wharton's jelly, offers the greatest number of harvestable mesenchymal stem cells. The review analyzed 161 studies reporting mesenchymal stem cell yields from various tissue sources, including adipose tissue, bone marrow, umbilical cord tissue, and placental tissue. It found that yields from umbilical cord tissue ranged from 10,000 to 4,700,000 cells per milliliter, far exceeding yields from other sources. Adipose tissue provided the next highest yields, ranging from 4,737 to 1,550,000 cells per milliliter. Bone marrow yields ranged more widely from 1 to 317,400 cells per millil
Bone marrow and peripheral Hematopoietic stem cell collection and processing.KISHORE KUMAR
This document provides an overview of bone marrow and peripheral blood stem cell collection and processing for hematopoietic stem cell transplantation. It discusses the indications for transplant, sources of stem cells, mobilization techniques using growth factors and chemotherapy, collection methods for bone marrow and peripheral blood, processing steps including volume reduction, cryopreservation and storage, and thawing for infusion. The key steps in bone marrow collection, peripheral blood mobilization, apheresis, and processing the collected stem cells are outlined.
Umbilical cord mesenchymal stem cells (UC-MSCs) show potential advantages over mesenchymal stem cells (MSCs) from other sources for regenerative medicine applications. UC-MSCs display higher proliferation rates and expression of embryonic genes compared to adult MSCs. Transcriptomic analyses indicate UC-MSCs express genes related to development of multiple tissues including bone, liver, cardiovascular and neural systems. While UC-MSCs can differentiate into cell types of multiple lineages, their therapeutic impact is thought to be mainly due to their paracrine effects and immunomodulatory properties. UC-MSCs could have advantages for treating autoimmune and neurodegenerative diseases.
Hematopoetic stem cell transplantation by Dr.Kumarbhargav KaptanBhargav Kaptan
The document reviews the current status of hematopoietic stem cell transplantation, describing the types of stem cells used including embryonic, adult, induced pluripotent, and umbilical cord stem cells. It discusses sources of hematopoietic stem cells, the process of transplantation including donor evaluation and conditioning regimens, and indications for transplantation in both malignant and non-malignant conditions such as sickle cell anemia, thalassemia major, and immunodeficiency diseases. Outcomes of transplantation for diseases like Hodgkin's lymphoma are also reviewed.
Autologous bone marrow transplant involves harvesting a patient's own bone marrow stem cells, storing them, and later re-infusing them after high-dose chemotherapy or radiation treatment to destroy cancerous cells. The stem cells help repopulate the bone marrow and restore the immune system. Complications can include infections during the neutropenic phase, graft-versus-host disease, and mucositis. Long term effects may include secondary cancers or sterility. Autologous transplants are commonly used to treat blood cancers like lymphoma or multiple myeloma.
Generation of Clonal CRISPR/Cas9-edited Human iPSC Derived Cellular Models an...Thermo Fisher Scientific
This document describes a workflow for generating clonal CRISPR-edited human induced pluripotent stem cell (hiPSC) lines. Key aspects of the workflow include:
1) Developing a hiPSC line that stably expresses Cas9 to facilitate efficient genome editing.
2) Optimizing delivery methods for CRISPR/Cas9 editing tools and improving single cell clone survival and isolation using laminin-521 and StemFlex medium.
3) Applying the workflow to generate hiPSC lines carrying disease-relevant mutations and testing the cell models in assays, finding increased sensitivity to stress in models of Parkinson's disease.
Effect of stemregenin1 and sb431542 small molecules on ex vivo expansion of u...Liberty University (LU)
This study investigated the ex vivo expansion of umbilical cord blood hematopoietic stem cells (HSCs) using three-dimensional (3D) polyethersulfone (PES) nanofiber scaffolds and two small molecules, Stemregenin1 (SR1) and SB431542. Isolated HSCs were cultured in 3D PES scaffolds or 2D conditions with SR1 and/or SB431542 and growth factors. On days 5 and 10, the cells were analyzed for marker expression, colony formation ability, and gene expression of stemness genes. SR1 increased HSC expansion while SB431542 induced more lineage-committed cells. SR1 upregulated stemness genes and
This document summarizes research on developing the optimal protocol for decellularizing human-sized kidneys for transplantation. Two decellularization protocols were tested on sheep kidneys - Protocol 1 used 1% SDS only, while Protocol 2 used 1% Triton X-100 and 0.5% SDS. Both protocols completely decellularized the kidneys, but Protocol 2 better preserved the extracellular matrix. Kidneys treated with Protocol 1 showed fluid extravasation after transplantation in sheep, and the sheep expired within a day. In contrast, kidneys treated with Protocol 2 showed intact vasculature after transplantation, and the sheep could be monitored for 3 days without notable fluid accumulation. This study demonstrates the importance of well-pre
Allogeneic hematopoietic stem cell transplantation (allo HSCT) from an HLA-matched related donor provides the most potent anti-leukemic effect of any post-remission therapy in AML, as demonstrated by the lowest rates of relapse.
Graft vs leukemia plays and important role here.
Provides the best chance of long-term survival
This study analyzed a novel Wharton's jelly formulation to quantify growth factors, cytokines, hyaluronic acid, and extracellular vesicles. All samples passed sterility testing. The formulation was found to contain numerous growth factors including IGFBPs and PDGF-AA. It also contained cytokines associated with immunomodulation, wound healing, and regeneration. High levels of hyaluronic acid were detected. Particles in the size range of extracellular vesicles were also present, enclosed by membranes. The study demonstrates that Wharton's jelly contains various regenerative components that may help reduce inflammation and augment healing of musculoskeletal injuries.
Peripheral blood stem cell transplantation (PBSCT) involves collecting stem cells from a patient's bloodstream and later infusing them back into the patient after chemotherapy or radiation therapy. PBSCT has replaced bone marrow as the most common stem cell transplantation procedure. Stem cells are collected from the bloodstream using growth factors alone or with chemotherapy, and the minimum number needed for a safe transplant is 2 million CD34+ cells per kilogram of body weight. PBSCT results in faster recovery time compared to bone marrow transplants due to higher numbers of stem cells and T cells collected.
The study tested the effect of adding umbilical cord blood to cultures used in a clonogenic assay. Peripheral blood was collected from healthy individuals and mononuclear cells were isolated. Cultures were prepared with and without cord blood using methylcellulose media and cytokines. Cultures containing cord blood yielded significantly more hematopoietic colonies than cultures without cord blood after 14 days, as determined by a paired t-test. The addition of cord blood enhanced colony growth in the clonogenic assay.
This study evaluated whether the phenotype and therapeutic potency of umbilical cord tissue-derived mesenchymal stromal cells (UC-MSCs) were affected by cryopreservation and thawing. Specifically, the study compared cultured UC-MSCs to those freshly thawed after cryopreservation. Results showed that the immunophenotype and immunomodulatory and angiogenic properties of UC-MSCs were not impaired by cryopreservation and thawing, as demonstrated by in vitro and in vivo assays. This suggests that potency impairment related to cryopreservation can be avoided through the specific production process used for these UC-MSCs.
This study developed a protocol to efficiently generate hematopoietic stem and progenitor cells (HSPCs) from human induced pluripotent stem cells (iPSCs) with the goal of producing cells for clinical transplantation. An initial iPSC plating density of 70,000 cells per plate optimized generation of CD34hi/CD45lo progenitor cells. Additionally, brief hyperthermic treatment of native human CD34+ cells and iPSC-derived HSPCs was found to increase polarization of membrane domains and enhance homing and engraftment in a mouse model through increased migration towards bone marrow niches. These findings provide a potential approach for improving engraftment of iPSC-derived HSPCs for
This document summarizes the biology and potential clinical applications of mesenchymal stem cells (MSCs) derived from the Wharton's jelly of the umbilical cord. It discusses how MSCs were originally isolated from bone marrow but umbilical cord is now seen as a promising alternative source due to its abundance of MSCs and the non-invasive collection method. MSCs from Wharton's jelly (WJ-MSCs) share properties with bone marrow MSCs yet are considered more primitive. The review examines the multilineage potential and immunomodulatory abilities of WJ-MSCs and their emerging roles in treating cancer, graft-versus-host disease, and systemic lupus erythemat
Reprogramming to pluripotency is possible from adult cells of different tissues and species through the ectopic expression of defined factors. The generated induced Pluripotent Stem Cells (iPSCs) are relevant for various purposes, including disease modeling, drug or toxicity screening and autologous cell therapy. Over the last few years, increased efforts are being made to improve the reprogramming techniques, the efficiency and quality of the generated iPSCs, as well as to identify the best cell source to be reprogrammed. Cells derived from fetal tissues, such as amniotic fluid, placenta and umbilical cord, offer distinct advantages in terms of reprogramming compared to adult somatic cells. Importantly, fetal cells are more primitive, easily achievable in sufficient numbers and are devoid of any ethical concern. They show great plasticity, high proliferation rate, low immunogenity and absence of teratoma formation. Therefore, they can be reprogrammed much faster and more efficiently than adult cells. Here, we provide a comprehensive overview of the advantages of reprogramming fetal sources in comparison to other commonly used cell types.
Hematopoietic stem cell transplant (HSCT) involves transplanting hematopoietic stem cells to re-establish normal bone marrow function in patients with blood disorders or cancer. HSCT has become an established treatment for many malignant and non-malignant blood diseases. HSCT sources include bone marrow, peripheral blood, and umbilical cord blood. The transplant process involves stem cell collection, processing, conditioning chemotherapy, stem cell infusion, and recovery. Complications can include graft-versus-host disease. Matching HLA antigens between donor and recipient is important for transplant success, especially in allogeneic HSCT. Advances have improved outcomes, but further progress is still needed.
1) The document categorizes diseases treated with blood-forming stem cells into standard therapies, therapies in clinical trials, and experimental treatments. It distinguishes between allogeneic transplants using donor stem cells and autologous transplants using a patient's own stem cells.
2) Standard therapies include various forms of leukemia, lymphoma, anemias, immune disorders, and other blood disorders. Clinical trials cover neurologic injuries, autoimmune diseases, cardiovascular diseases, and gene therapy for inherited disorders. Experimental treatments are still being researched in labs and animals.
3) Clinical trials using mesenchymal stem cells from umbilical cord tissue focus on diseases like Alzheimer's, aplastic anemia, autism
Cell-Replacement Therapy with Stem Cells in Neurodegenerative DiseasesSararajputsa
This document summarizes research on using stem cell transplantation as a potential therapy for neurodegenerative diseases like Amyotrophic Lateral Sclerosis (ALS). It discusses how stem cell transplantation has shown effectiveness in animal models of ALS through mechanisms like cell replacement, neurotrophic factor release, and endogenous stem cell proliferation. Early clinical trials transplanting stem cells into ALS patients showed feasibility and no severe side effects. However, more research is still needed to fully understand stem cell mechanisms and maximize their therapeutic potential for neurodegenerative diseases.
Treatable Diseases Using Stem Cells - Cryobanks IndiaSararajputsa
1) The document categorizes diseases treated with blood-forming stem cells into standard therapies, therapies in clinical trials, and experimental treatments. It distinguishes between allogeneic transplants using donor stem cells and autologous transplants using a patient's own stem cells.
2) Standard therapies include various forms of leukemia, lymphoma, anemias, immune disorders, and other blood disorders. Clinical trials cover neurologic injuries, autoimmune diseases, cardiovascular diseases, and gene therapy for inherited disorders. Experimental treatments are still being researched in labs and animals.
3) Clinical trials using mesenchymal stem cells from umbilical cord tissue focus on diseases like Alzheimer's, aplastic anemia, autism
This literature review finds that umbilical cord tissue, specifically Wharton's jelly, offers the greatest number of harvestable mesenchymal stem cells. The review analyzed 161 studies reporting mesenchymal stem cell yields from various tissue sources, including adipose tissue, bone marrow, umbilical cord tissue, and placental tissue. It found that yields from umbilical cord tissue ranged from 10,000 to 4,700,000 cells per milliliter, far exceeding yields from other sources. Adipose tissue provided the next highest yields, ranging from 4,737 to 1,550,000 cells per milliliter. Bone marrow yields ranged more widely from 1 to 317,400 cells per millil
Bone marrow and peripheral Hematopoietic stem cell collection and processing.KISHORE KUMAR
This document provides an overview of bone marrow and peripheral blood stem cell collection and processing for hematopoietic stem cell transplantation. It discusses the indications for transplant, sources of stem cells, mobilization techniques using growth factors and chemotherapy, collection methods for bone marrow and peripheral blood, processing steps including volume reduction, cryopreservation and storage, and thawing for infusion. The key steps in bone marrow collection, peripheral blood mobilization, apheresis, and processing the collected stem cells are outlined.
Umbilical cord mesenchymal stem cells (UC-MSCs) show potential advantages over mesenchymal stem cells (MSCs) from other sources for regenerative medicine applications. UC-MSCs display higher proliferation rates and expression of embryonic genes compared to adult MSCs. Transcriptomic analyses indicate UC-MSCs express genes related to development of multiple tissues including bone, liver, cardiovascular and neural systems. While UC-MSCs can differentiate into cell types of multiple lineages, their therapeutic impact is thought to be mainly due to their paracrine effects and immunomodulatory properties. UC-MSCs could have advantages for treating autoimmune and neurodegenerative diseases.
Hematopoetic stem cell transplantation by Dr.Kumarbhargav KaptanBhargav Kaptan
The document reviews the current status of hematopoietic stem cell transplantation, describing the types of stem cells used including embryonic, adult, induced pluripotent, and umbilical cord stem cells. It discusses sources of hematopoietic stem cells, the process of transplantation including donor evaluation and conditioning regimens, and indications for transplantation in both malignant and non-malignant conditions such as sickle cell anemia, thalassemia major, and immunodeficiency diseases. Outcomes of transplantation for diseases like Hodgkin's lymphoma are also reviewed.
Autologous bone marrow transplant involves harvesting a patient's own bone marrow stem cells, storing them, and later re-infusing them after high-dose chemotherapy or radiation treatment to destroy cancerous cells. The stem cells help repopulate the bone marrow and restore the immune system. Complications can include infections during the neutropenic phase, graft-versus-host disease, and mucositis. Long term effects may include secondary cancers or sterility. Autologous transplants are commonly used to treat blood cancers like lymphoma or multiple myeloma.
Generation of Clonal CRISPR/Cas9-edited Human iPSC Derived Cellular Models an...Thermo Fisher Scientific
This document describes a workflow for generating clonal CRISPR-edited human induced pluripotent stem cell (hiPSC) lines. Key aspects of the workflow include:
1) Developing a hiPSC line that stably expresses Cas9 to facilitate efficient genome editing.
2) Optimizing delivery methods for CRISPR/Cas9 editing tools and improving single cell clone survival and isolation using laminin-521 and StemFlex medium.
3) Applying the workflow to generate hiPSC lines carrying disease-relevant mutations and testing the cell models in assays, finding increased sensitivity to stress in models of Parkinson's disease.
Effect of stemregenin1 and sb431542 small molecules on ex vivo expansion of u...Liberty University (LU)
This study investigated the ex vivo expansion of umbilical cord blood hematopoietic stem cells (HSCs) using three-dimensional (3D) polyethersulfone (PES) nanofiber scaffolds and two small molecules, Stemregenin1 (SR1) and SB431542. Isolated HSCs were cultured in 3D PES scaffolds or 2D conditions with SR1 and/or SB431542 and growth factors. On days 5 and 10, the cells were analyzed for marker expression, colony formation ability, and gene expression of stemness genes. SR1 increased HSC expansion while SB431542 induced more lineage-committed cells. SR1 upregulated stemness genes and
This document summarizes research on developing the optimal protocol for decellularizing human-sized kidneys for transplantation. Two decellularization protocols were tested on sheep kidneys - Protocol 1 used 1% SDS only, while Protocol 2 used 1% Triton X-100 and 0.5% SDS. Both protocols completely decellularized the kidneys, but Protocol 2 better preserved the extracellular matrix. Kidneys treated with Protocol 1 showed fluid extravasation after transplantation in sheep, and the sheep expired within a day. In contrast, kidneys treated with Protocol 2 showed intact vasculature after transplantation, and the sheep could be monitored for 3 days without notable fluid accumulation. This study demonstrates the importance of well-pre
Allogeneic hematopoietic stem cell transplantation (allo HSCT) from an HLA-matched related donor provides the most potent anti-leukemic effect of any post-remission therapy in AML, as demonstrated by the lowest rates of relapse.
Graft vs leukemia plays and important role here.
Provides the best chance of long-term survival
This study analyzed a novel Wharton's jelly formulation to quantify growth factors, cytokines, hyaluronic acid, and extracellular vesicles. All samples passed sterility testing. The formulation was found to contain numerous growth factors including IGFBPs and PDGF-AA. It also contained cytokines associated with immunomodulation, wound healing, and regeneration. High levels of hyaluronic acid were detected. Particles in the size range of extracellular vesicles were also present, enclosed by membranes. The study demonstrates that Wharton's jelly contains various regenerative components that may help reduce inflammation and augment healing of musculoskeletal injuries.
Peripheral blood stem cell transplantation (PBSCT) involves collecting stem cells from a patient's bloodstream and later infusing them back into the patient after chemotherapy or radiation therapy. PBSCT has replaced bone marrow as the most common stem cell transplantation procedure. Stem cells are collected from the bloodstream using growth factors alone or with chemotherapy, and the minimum number needed for a safe transplant is 2 million CD34+ cells per kilogram of body weight. PBSCT results in faster recovery time compared to bone marrow transplants due to higher numbers of stem cells and T cells collected.
The study tested the effect of adding umbilical cord blood to cultures used in a clonogenic assay. Peripheral blood was collected from healthy individuals and mononuclear cells were isolated. Cultures were prepared with and without cord blood using methylcellulose media and cytokines. Cultures containing cord blood yielded significantly more hematopoietic colonies than cultures without cord blood after 14 days, as determined by a paired t-test. The addition of cord blood enhanced colony growth in the clonogenic assay.
This study evaluated whether the phenotype and therapeutic potency of umbilical cord tissue-derived mesenchymal stromal cells (UC-MSCs) were affected by cryopreservation and thawing. Specifically, the study compared cultured UC-MSCs to those freshly thawed after cryopreservation. Results showed that the immunophenotype and immunomodulatory and angiogenic properties of UC-MSCs were not impaired by cryopreservation and thawing, as demonstrated by in vitro and in vivo assays. This suggests that potency impairment related to cryopreservation can be avoided through the specific production process used for these UC-MSCs.
This study developed a protocol to efficiently generate hematopoietic stem and progenitor cells (HSPCs) from human induced pluripotent stem cells (iPSCs) with the goal of producing cells for clinical transplantation. An initial iPSC plating density of 70,000 cells per plate optimized generation of CD34hi/CD45lo progenitor cells. Additionally, brief hyperthermic treatment of native human CD34+ cells and iPSC-derived HSPCs was found to increase polarization of membrane domains and enhance homing and engraftment in a mouse model through increased migration towards bone marrow niches. These findings provide a potential approach for improving engraftment of iPSC-derived HSPCs for
This document summarizes the biology and potential clinical applications of mesenchymal stem cells (MSCs) derived from the Wharton's jelly of the umbilical cord. It discusses how MSCs were originally isolated from bone marrow but umbilical cord is now seen as a promising alternative source due to its abundance of MSCs and the non-invasive collection method. MSCs from Wharton's jelly (WJ-MSCs) share properties with bone marrow MSCs yet are considered more primitive. The review examines the multilineage potential and immunomodulatory abilities of WJ-MSCs and their emerging roles in treating cancer, graft-versus-host disease, and systemic lupus erythemat
Reprogramming to pluripotency is possible from adult cells of different tissues and species through the ectopic expression of defined factors. The generated induced Pluripotent Stem Cells (iPSCs) are relevant for various purposes, including disease modeling, drug or toxicity screening and autologous cell therapy. Over the last few years, increased efforts are being made to improve the reprogramming techniques, the efficiency and quality of the generated iPSCs, as well as to identify the best cell source to be reprogrammed. Cells derived from fetal tissues, such as amniotic fluid, placenta and umbilical cord, offer distinct advantages in terms of reprogramming compared to adult somatic cells. Importantly, fetal cells are more primitive, easily achievable in sufficient numbers and are devoid of any ethical concern. They show great plasticity, high proliferation rate, low immunogenity and absence of teratoma formation. Therefore, they can be reprogrammed much faster and more efficiently than adult cells. Here, we provide a comprehensive overview of the advantages of reprogramming fetal sources in comparison to other commonly used cell types.
Hematopoietic stem cell transplant (HSCT) involves transplanting hematopoietic stem cells to re-establish normal bone marrow function in patients with blood disorders or cancer. HSCT has become an established treatment for many malignant and non-malignant blood diseases. HSCT sources include bone marrow, peripheral blood, and umbilical cord blood. The transplant process involves stem cell collection, processing, conditioning chemotherapy, stem cell infusion, and recovery. Complications can include graft-versus-host disease. Matching HLA antigens between donor and recipient is important for transplant success, especially in allogeneic HSCT. Advances have improved outcomes, but further progress is still needed.
1) The document categorizes diseases treated with blood-forming stem cells into standard therapies, therapies in clinical trials, and experimental treatments. It distinguishes between allogeneic transplants using donor stem cells and autologous transplants using a patient's own stem cells.
2) Standard therapies include various forms of leukemia, lymphoma, anemias, immune disorders, and other blood disorders. Clinical trials cover neurologic injuries, autoimmune diseases, cardiovascular diseases, and gene therapy for inherited disorders. Experimental treatments are still being researched in labs and animals.
3) Clinical trials using mesenchymal stem cells from umbilical cord tissue focus on diseases like Alzheimer's, aplastic anemia, autism
Cell-Replacement Therapy with Stem Cells in Neurodegenerative DiseasesSararajputsa
This document summarizes research on using stem cell transplantation as a potential therapy for neurodegenerative diseases like Amyotrophic Lateral Sclerosis (ALS). It discusses how stem cell transplantation has shown effectiveness in animal models of ALS through mechanisms like cell replacement, neurotrophic factor release, and endogenous stem cell proliferation. Early clinical trials transplanting stem cells into ALS patients showed feasibility and no severe side effects. However, more research is still needed to fully understand stem cell mechanisms and maximize their therapeutic potential for neurodegenerative diseases.
Treatable Diseases Using Stem Cells - Cryobanks IndiaSararajputsa
1) The document categorizes diseases treated with blood-forming stem cells into standard therapies, therapies in clinical trials, and experimental treatments. It distinguishes between allogeneic transplants using donor stem cells and autologous transplants using a patient's own stem cells.
2) Standard therapies include various forms of leukemia, lymphoma, anemias, immune disorders, and other blood disorders. Clinical trials cover neurologic injuries, autoimmune diseases, cardiovascular diseases, and gene therapy for inherited disorders. Experimental treatments are still being researched in labs and animals.
3) Clinical trials using mesenchymal stem cells from umbilical cord tissue focus on diseases like Alzheimer's, aplastic anemia, autism
Prashant Kumar has over 10 years of experience in JDE, including 9 years of experience providing support for JDE ERP implementations. He has extensive expertise in JDE modules including Inventory, Sales, Transportation, and Procurement. He has led teams and taken on project lead roles for various large clients such as Tupperware, Lafarge, and KPIT.
Balunga Mbolekwa provides his Curriculum Vitae to the City of Tshwane. He has worked as a system operator for the City of Tshwane since 2006 where he uses programs like Microsoft Office, SAP and Suprima to manage client databases and address queries. He also previously worked as a cashier and sales assistant at Woolworths from 2004 to 2006. Balunga has a diploma in media and journalism and matriculated from high school in 2003.
Advances in Stem Cell Therapy for Erectile DysfunctionSararajputsa
Stem Cell therapy for erectile dysfunction has been investigated in 35 published studies, with one being a small-scale clinical trial. Out of these 35 studies, 19 are concerned with cavernous nerve (CN) injury-associated ED while 10 with diabetes mellitus- (DM-) associated ED. For more details visit: http://www.cryobanksindia.com/moms-corner/case-studies/
Stem Cell Therapy for Kidney DysfunctionSararajputsa
This review article summarizes recent advances in regenerative medicine for the kidney. It discusses four main strategies: (1) identifying renotropic growth factors that promote kidney repair; (2) identifying renal stem/progenitor cells in embryonic and adult kidneys; (3) using cell therapies with bone marrow-derived cells like hematopoietic stem cells and mesenchymal stem cells; and (4) reconstructing artificial kidneys or renal components using embryonic stem cells or induced pluripotent stem cells. The article examines evidence for each strategy and highlights ongoing work to understand renotropic signaling pathways and characterize renal stem/progenitor cells, with the goal of developing effective regenerative therapies for treating kidney disease
1) The document categorizes diseases treated with blood-forming stem cells into standard therapies, therapies in clinical trials, and experimental treatments. It distinguishes between allogeneic transplants using donor stem cells and autologous transplants using a patient's own stem cells.
2) Standard therapies include various forms of leukemia, lymphoma, anemias, immune disorders, and other blood disorders. Clinical trials cover neurologic injuries, autoimmune diseases, cardiovascular diseases, and gene therapy for inherited disorders. Experimental treatments are still being researched in labs and animals.
3) Clinical trials using mesenchymal stem cells from umbilical cord tissue focus on diseases like Alzheimer's, aplastic anemia, autism
Prashant Kumar has over 10 years of experience in JDE, including 9 years of experience providing support for JDE ERP implementations. He has extensive expertise in JDE modules including Inventory, Sales, Transportation, and Procurement. He has led implementation projects for multiple clients and was awarded an ICON award in 2011 for his work.
- Babies delivered after 40 weeks gestation did not have lower sample volumes, stem cell counts, or viability compared to those delivered before 40 weeks. There was no statistical difference found.
- Increasing maternal age did not correlate with declines in sample volume, viability, or CD34+ stem cell count. Statistical tests found no significant differences between age groups.
- The hypotheses that gestation periods over 40 weeks or increased maternal age would negatively impact cord blood sample quality were not supported by the data analysis. The null hypotheses could not be rejected.
In placental mammals, the umbilical cord (also called the navel string, birth cord or funiculus umbilicalis) is a conduit between the developing embryo or fetus and the placenta. During prenatal development, the umbilical cord is physiologically and genetically part of the fetus and, (in humans), normally contains two arteries (the umbilical arteries) and one vein (the umbilical vein), buried within Wharton's jelly. The umbilical vein supplies the fetus with oxygenated, nutrient-rich blood from the placenta. Conversely, the fetal heart pumps deoxygenated, nutrient-depleted blood through the umbilical arteries back to the placenta.
The blood within the umbilical cord, known as cord blood, is a rich and readily available source of primitive, undifferentiated stem cells (of type CD34-positive and CD38-negative). These cord blood cells can be used for bone marrow transplant.
Some parents choose to have this blood diverted from the baby's umbilical blood transfer through early cord clamping and cutting, to freeze for long-term storage at a cord blood bank should the child ever require the cord blood stem cells (for example, to replace bone marrow destroyed when treating leukemia).
In the future, cord blood-derived embryonic-like stem cells (CBEs) may be banked and matched with other patients, much like blood and transplanted tissues. The use of CBEs could potentially eliminate the ethical difficulties associated with embryonic stem cells (ESCs).
CD34+ cells were isolated from the PLC/PRF/5 hepatoma cell line. These CD34+ cells appeared to function as liver cancer stem cells (LCSCs) based on their ability to form human liver carcinomas (HLCs) in immunodeficient mice with as few as 100 cells injected. When various subpopulations of these CD34+ PLC cells were injected into mice, they generated three types of HLCs - hepatocellular carcinomas (HCCs), cholangiocarcinomas (CCs), and combined hepatocellular cholangiocarcinomas (CHCs). The expression of different cell surface antigens like OV6 and CD31 on CD
The document summarizes current trends in cell therapy, including:
1) Attempts to expand hematopoietic stem cells for clinical use have not achieved long-term engraftment, though total CD34+ cells are expanded with benefits.
2) Cord blood banking trends involve improving quality over quantity, and cord blood may be used for non-hematological diseases.
3) Over 3000 patients have been treated safely for heart diseases with cell therapy, and the field is growing rapidly though mechanisms are better understood as trophic rather than differentiation.
This document describes a label-free method for cell counting using paramagnetic bead aggregation. When cells are lysed in a chaotropic solution, the released DNA causes paramagnetic beads to aggregate. The extent of aggregation correlates with the amount of DNA and cell number. This allows direct enumeration of cells from crude samples. The method is demonstrated by monitoring bacterial growth and obtaining white blood cell counts from whole blood samples, showing good agreement with standard methods. Specific cell types like CD4+ T cells can also be enumerated using bead-based immunocapture prior to the aggregation step. The method requires only inexpensive equipment and could provide an accessible alternative to more expensive cell counting techniques.
This study analyzed the use of the biosimilar granulocyte colony-stimulating factor (GCSF) Ratiograstim for peripheral blood stem cell mobilization and engraftment compared to the originator GCSF Neupogen. Data were collected for 154 patients mobilized with Ratiograstim and 131 historical controls mobilized with Neupogen. There were no statistically significant differences between the two groups in CD34+ stem cell collection parameters, number of days required for collection, or time to engraftment. This represents one of the largest direct comparisons of a biosimilar GCSF to the originator for stem cell mobilization, and found no differences, suggesting biosimilar GCSF can be used
This document describes a new bioartificial pancreas (BAP) device called "b-Air" that is designed to overcome challenges with islet transplantation, specifically inadequate oxygen supply and protection from the host immune system. The b-Air consists of islets immobilized in an alginate hydrogel within a gas chamber that can be supplied with oxygen via subdermal ports. In animal studies, the b-Air completely normalized blood glucose and HbA1c levels in diabetic rats for up to 6 months. The device was dependent on oxygen supply and provided immune protection by allowing survival of both isografts and allografts. Explanted devices showed intact, functional islets without inflammation. Increased islet loading
Interventional orthopedics foundation amniotic tissue products posterChris Centeno
This document evaluates four placental tissue-derived products - AmnioFix Injectable, BioDRestore, FloGraS Freedom, and Ovation Cellular Repair Matrix - to determine if they contain plastic-adherent, multipotent stem cells. Through microscopy, flow cytometry, and cell culture experiments, the researchers found no evidence of expandable cell cultures from the products, though some increased cellular proliferation and reactive oxygen species levels were observed with AmnioFix and Ovation.
This study evaluated a novel immunoisolating membrane system for transplanting rat pancreatic islets (xenografts) into diabetic minipigs without immunosuppressive therapy. Rat islets were encapsulated in alginate and placed in a macrochamber covered by a poly-membrane. The chamber had a gas system to supply oxygen to the islets. Diabetic minipigs received the transplants and were monitored for up to 90 days. The rat islets functioned persistently and restored normoglycemia in the minipigs without immunosuppressants, demonstrating the potential of this system for treating diabetes with xenogeneic islet transplantation without drug-based immunosuppression.
Enhancing Limb Salvage by Non-Mobilized Peripheral Blood Angiogenic Cell Prec...lifextechnologies
This document describes a study that assessed the efficacy and safety of implanting non-mobilized peripheral blood angiogenic cell precursors (NMPB-ACPs) in 6 patients with critical limb ischemia who were not candidates for standard revascularization treatments. The results showed no complications from the procedure. Five patients (83.3%) had improved circulation in the distal limb, with complete healing of ulcers/amputation sites. However, one patient later required an amputation due to foot infection. The preliminary results suggest NMPB-ACPs therapy may be safe and improve circulation, but a larger controlled trial is needed to confirm these findings.
Efficacy of liquid based cytology versus conventional smearsAnamika Dev
This study compares the efficacy of liquid-based cytology (LBC) to conventional smear methods for fine needle aspiration cytology samples. The study analyzed 110 cases of various lesions collected over 2 years. Results showed that LBC produced more clear backgrounds and better preserved and dispersed cells compared to conventional smears. LBC allowed for improved diagnosis of lesions like ductal carcinoma, lymph node metastases, and bone lesions. However, LBC required more processing and had some artifacts that could complicate interpretation. Overall, LBC improved sample quality and reduced inadequate sampling compared to conventional smears.
Current and Future Applications of Umbilical Cord Blood Cell TransfusionIvan Seah
Umbilical cord blood transfusion is currently used to treat blood-related diseases like leukemia, lymphoma, and inherited blood disorders. It has advantages over bone marrow transplants in having a longer shelf life, lower risk of graft-versus-host disease, and better HLA matching. Studies have shown umbilical cord blood transplants can successfully treat diseases like Hurler's Syndrome. Future applications may include using umbilical cord blood cells for organ regeneration, treating autoimmune diseases, and gene therapy. However, challenges remain in increasing stem cell counts for adult diseases and reducing risks of new gene therapies.
Involvement of nitric oxide and the ovarian blood follicle barrier in murine ...Rashmi Nemade
Chronic treatment with hCG induced enlarged ovaries containing multiple follicular cysts in immature mice. These cysts had few granulosa cells, secreted high levels of testosterone, and had impaired blood follicle barrier function. Inhibiting nitric oxide synthesis using L-NAME during cyst formation reduced cyst size, maintained normal testosterone levels, and preserved blood follicle barrier reactivity in cystic follicles. This suggests nitric oxide is involved in the formation of hCG-induced murine follicular cysts and their associated complications.
1. Researchers developed a protocol to efficiently generate human hematopoietic stem and progenitor cells (HSPCs) from induced pluripotent stem cells (iPSCs). An initial iPSC plating density of 70,000 cells per plate produced the desired CD34hi/CD45lo HSPC population.
2. A brief hyperthermic treatment of 39.5°C was found to enhance the homing of native human CD34+ cells within the bone marrow of mice compared to untreated cells. This increased homing may be due to greater polarization of membrane domains enriched in adhesion molecules.
3. The findings provide preliminary evidence that hyperthermic treatment could enhance the homing and engra
Overcoming the challenges of molecular diagnostics in government health insti...Yakubu Sunday Bot
overcoming the challenges of molecular diagnostics in government owned health institution in nigeria.Several challenges abound in the Nigerian health sector ranging from financial,political and lack of commitment.Its obvious and no wonder the state of health care deliveryy, vis a vis its quality of care to its citizenry.
Scientists screened 18 novel compounds for their ability to kill cancer cells. Several compounds showed potent cytotoxic effects, with IC50 values less than 1 μM on multiple cell lines. Three compounds - TMCOS-3, TMCOS-6, and TMCOS-11 - were found to induce apoptotic cell death through DNA fragmentation and caspase activation. TMCOS-11 was found to specifically inhibit tubulin polymerization and cause cell cycle arrest in the G2/M phase. These findings suggest that some of the compounds may be promising new anti-cancer drugs that work by targeting the microtubule protein tubulin.
The application of nanotechnology, microfluidics, bioreactors with kidney cells, and miniaturized sorbent systems to regenerate dialysate makes clinical reality seem closer than ever before. Finally, stem cells hold much promise, But more far realistic. In summary, nephrology is at an exciting crossroad with the application of innovative and novel technologies to RRT that hold considerable promise for the near future. Bioartificial Kidney as an ideal form of RRT would mimic the functions of natural kidneys and be affordable to the patient
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Administration of Autologous Bone Marrow Stem Cells Into Spinal Cord Injury P...◂ Justin (M) Gaines ▸
This document summarizes a study that administered autologous bone marrow stem cells (BMSCs) via multiple routes (directly into the spinal cord, directly into the spinal canal, and intravenous) to 8 spinal cord injury patients. The study found that administering BMSCs via multiple routes was safe and improved patients' quality of life based on evaluations using scales like ASIA, Barthel, Frankel, and a new bladder function scale. To date, administering BMSCs to 52 spinal cord injury patients has had no cases of tumor formation, infection, or increased pain and few minor adverse events.
Similar to Comparison on Techniques of Cell Seperation (20)
Michigan HealthTech Market Map 2024. Includes 7 categories: Policy Makers, Academic Innovation Centers, Digital Health Providers, Healthcare Providers, Payers / Insurance, Device Companies, Life Science Companies, Innovation Accelerators. Developed by the Michigan-Israel Business Accelerator
Empowering ACOs: Leveraging Quality Management Tools for MIPS and BeyondHealth Catalyst
Join us as we delve into the crucial realm of quality reporting for MSSP (Medicare Shared Savings Program) Accountable Care Organizations (ACOs).
In this session, we will explore how a robust quality management solution can empower your organization to meet regulatory requirements and improve processes for MIPS reporting and internal quality programs. Learn how our MeasureAble application enables compliance and fosters continuous improvement.
At Apollo Hospital, Lucknow, U.P., we provide specialized care for children experiencing dehydration and other symptoms. We also offer NICU & PICU Ambulance Facility Services. Consult our expert today for the best pediatric emergency care.
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TEST BANK For Accounting Information Systems, 3rd Edition by Vernon Richardso...rightmanforbloodline
TEST BANK For Accounting Information Systems, 3rd Edition by Vernon Richardson, Verified Chapters 1 - 18, Complete Newest Version
TEST BANK For Accounting Information Systems, 3rd Edition by Vernon Richardson, Verified Chapters 1 - 18, Complete Newest Version
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Trauma Outpatient Center is a comprehensive facility dedicated to addressing mental health challenges and providing medication-assisted treatment. We offer a diverse range of services aimed at assisting individuals in overcoming addiction, mental health disorders, and related obstacles. Our team consists of seasoned professionals who are both experienced and compassionate, committed to delivering the highest standard of care to our clients. By utilizing evidence-based treatment methods, we strive to help our clients achieve their goals and lead healthier, more fulfilling lives.
Our mission is to provide a safe and supportive environment where our clients can receive the highest quality of care. We are dedicated to assisting our clients in reaching their objectives and improving their overall well-being. We prioritize our clients' needs and individualize treatment plans to ensure they receive tailored care. Our approach is rooted in evidence-based practices proven effective in treating addiction and mental health disorders.
Under Pressure : Kenneth Kruk's StrategyKenneth Kruk
Kenneth Kruk's story of transforming challenges into opportunities by leading successful medical record transitions and bridging scientific knowledge gaps during COVID-19.
Healthy Eating Habits:
Understanding Nutrition Labels: Teaches how to read and interpret food labels, focusing on serving sizes, calorie intake, and nutrients to limit or include.
Tips for Healthy Eating: Offers practical advice such as incorporating a variety of foods, practicing moderation, staying hydrated, and eating mindfully.
Benefits of Regular Exercise:
Physical Benefits: Discusses how exercise aids in weight management, muscle and bone health, cardiovascular health, and flexibility.
Mental Benefits: Explains the psychological advantages, including stress reduction, improved mood, and better sleep.
Tips for Staying Active:
Encourages consistency, variety in exercises, setting realistic goals, and finding enjoyable activities to maintain motivation.
Maintaining a Balanced Lifestyle:
Integrating Nutrition and Exercise: Suggests meal planning and incorporating physical activity into daily routines.
Monitoring Progress: Recommends tracking food intake and exercise, regular health check-ups, and provides tips for achieving balance, such as getting sufficient sleep, managing stress, and staying socially active.
Can Allopathy and Homeopathy Be Used Together in India.pdfDharma Homoeopathy
This article explores the potential for combining allopathy and homeopathy in India, examining the benefits, challenges, and the emerging field of integrative medicine.
Hypertension and it's role of physiotherapy in it.Vishal kr Thakur
This particular slides consist of- what is hypertension,what are it's causes and it's effect on body, risk factors, symptoms,complications, diagnosis and role of physiotherapy in it.
This slide is very helpful for physiotherapy students and also for other medical and healthcare students.
Here is summary of hypertension -
Hypertension, also known as high blood pressure, is a serious medical condition that occurs when blood pressure in the body's arteries is consistently too high. Blood pressure is the force of blood pushing against the walls of blood vessels as the heart pumps it. Hypertension can increase the risk of heart disease, brain disease, kidney disease, and premature death.
LGBTQ+ Adults: Unique Opportunities and Inclusive Approaches to CareVITASAuthor
This webinar helps clinicians understand the unique healthcare needs of the LGBTQ+ community, primarily in relation to end-of-life care. Topics include social and cultural background and challenges, healthcare disparities, advanced care planning, and strategies for reaching the community and improving quality of care.
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This particular slides consist of- what is Pneumothorax,what are it's causes and it's effect on body, risk factors, symptoms,complications, diagnosis and role of physiotherapy in it.
This slide is very helpful for physiotherapy students and also for other medical and healthcare students.
Here is a summary of Pneumothorax:
Pneumothorax, also known as a collapsed lung, is a condition that occurs when air leaks into the space between the lung and chest wall. This air buildup puts pressure on the lung, preventing it from expanding fully when you breathe. A pneumothorax can cause a complete or partial collapse of the lung.
1. International Journal of Stem Cells Vol. 3, No. 1, 2010
ORIGINAL ARTICLE
32
Accepted for publication March 27, 2010
Correspondence to Colin McGuckin
CTI-LYON, Cell Therapy Research Institute, Parc Technologique
de Lyon-St Priest, Cèdre 1, 97 Allée Alexandre Borodine, 69800
SAINT PRIEST LYON, France
Tel: +33-4-26 03 01 29, Fax: +33-4-26 03 01 29
E-mail: c.mcguckin@conoworld.com
The Cord Blood Separation League Table: a Comparison of
the Major Clinical Grade Harvesting Techniques for
Cord Blood Stem Cells
Christina Basford
1
, Nicolas Forraz
2
, Saba Habibollah
1
, Kendal Hanger
1
, Colin McGuckin
2
1
Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, NE1 3BZ,
United Kingdom, 2
CTI-LYON, Cell Therapy Research Institute, Parc Technologique de Lyon-St Priest, Cèdre 1,
97 Allée Alexandre Borodine, 69800 SAINT PRIEST LYON, France
Background and Objectives: Well over 1 million Umbilical Cord Blood units (UCB) have been stored globally in the
last 10 years. Already, over 20,000 transplants been performed using UCB for haematopoietic reconstitution alone,
now this potential is joined by regenerative medicine. However, more needs to be known about processing of this
stem cell source for it to reach full potential.
Methods and Results: In this study we evaluated five separation methods: plasma depletion, density gradient,
Hetastarch, a novel method known as PrepaCyte-CB and an automated centrifugal machine. Sepax gives the highest
recovery of nucleated cells, an average of 78.8% (SD±21.36). When looking at CD34+ haematopoietic stem cells
PrepaCyte-CB provided the greatest recovery at 74.47% (SD±8.89). For volume reduction density gradient was the
most effective leaving 0.03×10
6
RBC/ml, 8 times more efficient than its nearest competitor PrepaCyte-CB (p<0.05).
Finally PrepaCyte-CB processing left samples with the highest clonogenic potential after processing and more sig-
nificantly after cryopreservation: 9.23 CFU/108
cells (SD±2.33), 1.5 fold more effective than its nearest rival Sepax
(p<0.05).
Conclusions: PrepaCyte-CB was the most flexible method; the only processing type unaffected by volume. Results in-
dicate that processing choice is important depending on your final intended use.
Keywords: Stem cells, Bioprocessing, Umbilical cord blood, Transplantation
Introduction
The global rise in Umbilical Cord Blood (UCB) as a
transplant source, has been amazing, over 20,000 trans-
plants already having taken place in haematopoietic re-
constructions alone (1). It has become a real alternative
to bone marrow (BM) and peripheral blood (PB) as a
source of adult stem cells to treat multiple diseases. The
first properly validated transplant took place in Paris in
1989 and was performed by Gluckman et al. They were
successful in reconstituting the haematopoietic system of
a child with Fanconi’s anaemia with UCB from an HLA-
identical sibling rather than BM (2). However, we believe
the success of UCB for transplantation can be traced back
to 1970, when a young male with acute leukaemia, re-
ceived a multiple (eight) cord blood unit transplant, re-
maining diseases free for twelve months After the triumph
of these early infusions of UCB more than 85 conditions
can currently be treated using this stem cell source, such
as the previously mentioned Fanconi’s anaemia, a BM fail-
ure disorder (2), metabolic disorders like Krabbe’s disease
(3) and immune defects like severe combined immune de-
2. Christina Basford, et al: The Cord Blood Separation League Table 33
ficiency (SCID) (4).
UCB has become such a popular adult stem cell source
for many reasons, not least because over 130 million births
worldwide per annum represents the largest, easily avail-
able stem cell source. It also allows for storage of units
from ethnic minorities not easily possible within BM reg-
istries (5). This potentially allows for an increase in the
rate of matched unrelated donor allogeneic transplants (6).
It has also been found that there is a lower risk of graft
versus host disease (GvHD) when transplanting UCB
when compared to BM (7). This could be due to the fact
that the cells transplanted from UCB are more naïve and
have lower human leukocyte antigen (HLA) protein ex-
pression (6). UCB has been shown to contain a higher fre-
quency of early progenitor cells than PB or BM (8). Fur-
ther findings show that term and pre-term UCB contain
significantly higher number of early and committed pro-
genitor cells, and that they are better able to form col-
ony-forming-unit granulocyte-macrophage (CFU-GM) when
compared to adult PB (9). Other studies show that as well
as being a source of haematopoietic progenitor cells, UCB
also contains non-haematopoietic stem or progenitor cells
including mesenchymal and endothelial precursors (10).
Even more recently UCB is becoming a real player in
the regenerative medicine field. There are many groups
looking to develop tissues for either transplantation or
drug testing with many successes. For example, the Mc-
Guckin and Denner groups, working in Texas, have suc-
ceeded in being able to generate insulin producing islet
cells from UCB with the hope to be able to treat diabetes
in the future (11). Another group have managed to pro-
duce functioning neural cells (12) showing great potential
in the treatment of neurological disorders.
So, although there are many benefits to using UCB for
transplantation there is one major limitation: the total
nucleated cell (TNC) count and cell number recoverable
from a single unit. This is affected by unit size, maternal
factors such as number of previous pregnancies and age
of mother (13), limited volumes available from each sam-
ple, but not least the processing method used. Together,
these factors highlight the need to make processing as effi-
cient as possible (14) to make cord blood banking a real
option. Many methods are currently available and the pur-
pose of this study was to evaluate five of the main types.
Techniques were varied; from density gradient separation,
like Lymphoprep or Ficoll-Paque (15), rouleaux formation
using Hetastarch (HES), a starch based method causing
red cells to clump (16), plasma depletion; a simple volume
reduction method which avoids the addition of any chem-
icals and simply removes the plasma (17), a novel closed
separation kit, closed separation kit, known as PrepaCyte-
CB which offers rapid and specific cell separation (18).
In this study we used a PrepaCyte-CB kit designed specifi-
cally for UCB units, to allow recovery of other non-eryth-
roid subsets such as leukocytes, thrombocytes and stem
cells, including both hematopoietic stem cells (HSC) and
multi-lineage progenitor cells, without them needing to be
bound to unwanted chemicals (19) or particles. The final
separation system chosen was also the only fully auto-
mated system, provided by Biosafe, known as the ‘Sepax’
machine (20).
Materials and Methods
Inclusion criteria
Samples were collected from the Royal Victoria Infir-
mary Maternity Unit, Newcastle upon Tyne, UK, only af-
ter written and informed consent was gained from the
parents. The protocol followed, was reviewed and ap-
proved by the National Health Service local ethics com-
mittee. A negative viral profile and infection status was
required. UCB units were processed and cryopreserved
within 24 hours of collection and only if the sample ex-
ceeded 40 ml.
Collection
For logistical and reasons of homogeneity of treatment,
samples were only collected from caesarean section births.
Collection took place post partum, after the placenta was
delivered. The cord was clamped in line with currently
used protocols at the hospital, and was performed by the
surgeon. Under no circumstance was the birthing proce-
dure adjusted or changed due to our study. Two clamps
were used: one placed close to the placenta and one close
to the baby. The placenta was then hung in a cone shaped
collection vessel with the cord hanging through the un-
derside. Collection bags used contain citrate phosphate
dextrose adenine (CPD-A) anticoagulant and had a needle
attached (Baxter PL146-CPDA-1-35 ml Deerfield IL, USA).
This was spiked into the bottom of the cord allowing the
blood to drain into the collection bag. Only blood from
the umbilical cord vein was collected, not from the pla-
centa. Gravity and natural movement were used to drain
the cord. Once sample collection was complete, the unit
was transported back to the laboratory and stored at room
temperature until processing was initiated.
Processing
All umbilical cord blood samples used in this study
were processed within 24 hours of collection. For each
3. 34 International Journal of Stem Cells 2010;3:32-45
unit, analysis samples were taken pre and post processing
and post cryopreservation/thaw.
Hetastarch (HES): The UCB was transferred to 150 ml
transfer bags (Baxter, R4R2001) then HES (Baxter B5084
6% in 0.9% Sodium Chloride) was added to the UCB at
a concentration of 20% blood volume. The bag was centri-
fuged at 125×g for 10 min, with the brake off, (Jouan
CR422. St-herblain, France), to stop disruption of the
RBC pellet. Using a plasma expresser (Fenwal BM-1. Lake
Zurich IL, USA), the supernatant containing the desired
nucleated cells was expressed off into a second transfer
bag. The second bag was then centrifuged at 400 to 500×g
for 10 min (Jouan CR422). Again using a plasma ex-
presser, the supernatant was removed into a third transfer
bag and this time discarded, leaving the pelleted nucleated
cells in the second bag, where they were then resuspended
in Human Serum Albumin (HSA) (Bio Products Labora-
tory PL08801/006. Elstree, UK).
PrepaCyte-CB: Before adding the UCB unit to the
PrepaCyte-CB device it was thoroughly mixed using a
plate rocker (Genesis Blood Collection Mixer CM-735.
Hackensack NJ, USA). The UCB unit was then spiked
with the connecting tube from the PrepaCyte-CB system
(www.BioE.com) allowing the blood to drain into it. For
optimal recovery it was recommended to drain a portion
of the reagent-cord blood mixture back into the collection
bag, mix and transfer bag into the processing bag. The
tubing between UCB collection bag and the bag set was
heat sealed and the collection bag discarded. The bag set
containing the reagent-blood mixture was rocked for 3∼5
min, 15∼20 rocks per min. After mixing, the bag set was
hung on a plasma expresser (Fenwal BM-1) for 30 min
to allow the unwanted cells to aggregate and sediment.
After sedimentation, using the plasma expresser, the TNC-
rich supernatant was pushed into the next bag for cen-
trifugation at 400 to 500×g for 10 min, with a low break
to avoid disruption of the pellet (Jouan CR422). After cen-
trifugation the TNC and stem cell portion was pelleted
allowing the unwanted second supernatant to drain back
through the system into the first bag with the unwanted
RBC portion of the sample. The stem cell fraction then
continued into the cryopreservation bag or equally it could
be used in the laboratory for tissue culture purposes.
Plasma depletion: The collection bag was connected to
a 300 ml transfer bag (Baxter) and the product flowed
through. It was then heat sealed and the tubing and collec-
tion bag were removed. The bag was spun for 10 min at
2,400 rpm (temp. 15∼26o
C). The bag was carefully re-
moved from the centrifuge and hung on a plasma ex-
pressor, where the unwanted plasma drained into a 150
ml transfer bag (Baxter). After removal of plasma, the
bags were heat sealed and removed. The UCB unit was
thoroughly mixed to resuspend the cells in the residual
plasma. The cryobag was then attached, using sterile tech-
nique and the UCB was allowed to flow into it. The un-
wanted bag and tubing was then heat sealed and removed.
Biosafe’s sepax: This fully automated, functionally
closed and sterile system, was controlled by computer
software. It was possible to isolate the HSC rich buffy-coat
of a UCB unit to a final volume of 10 to 90 ml. Each unit
was separated as per the user guide with a single use kit,
which was inserted into the machine. Briefly, the UCB
was added to the machine where it filled the central
chamber. Whilst filling, the system simultaneously sedi-
mented the UCB unit. The sample was spun at speeds of
up to 1,900×g and the blood components were separated
concentrically. Then, using optical sensors and motorised
stopcocks the blood components were directed to indivi-
dual blood bags and extracted from the UCB unit (http://
wwwbiosafe.ch). This method was recently validated by
the Besançon Cord Blood Bank in France (20).
Density gradient; ficoll-paque/lymphoprep: Ficoll is
a neutral, highly branched, high-mass, hydrophilic poly-
saccharide which can readily dissolve in aqueous solu-
tions. The UCB was diluted 50:50 with a buffered saline
solution (PBS) and was then layered over the Ficoll. The
sample was centrifuged at 800×g, for 20 min, to accelerate
the density gradient separation, as per manufacturers
guide (http://www.axis-shield-density-gradient-media. com).
The buffy layer of WBCs was at the interface between the
sample and the medium, and was collected using a sterile
Pasteur pipette. The buffy coat was then washed with
PBS.
Sample analysis
Enumeration by differential cell counting: The cell
counts were performed using the CellDyn4000 Analyser,
a mechanical method. A sample of 1 ml was applied to
the machine at each of the three points during processing.
The WBC count and differential determine the number
of white blood cells and the percentage of each type of
WBC in each unit of blood.
Colony forming unit assays: A cell suspension was pre-
pared with the appropriate number of cells; at 0.25×
105
/plate. Colony counts must then be multiplied by 4 to
give the number of colonies per 105
cells. The relevant
number of cells was added to make up a volume of IMDM
(Gibco/Invitrogen 21980-032. Paisley, UK)+20% FCS+
supplements to 0.5 ml, then added to a 3.5 ml aliquot of
methylcellulose culture medium, (Stem cell Technologies
4. Christina Basford, et al: The Cord Blood Separation League Table 35
Table 1. Antibody cocktails used for cell separation assessment: flow cytometry
Antibody Type Volume (μl) Cat. no. Distributor Tube no.
CD45 (APC Cy7) IgG1 5 557833 BD Biosciences 1, 2, 3, 4
CD235a (PE) IgG2b 5 555570 BD Pharmingen 1, 2, 3, 4
7AAD IgG1 5 559925 BD Pharmingen 1, 2, 3, 4
CD14 (Pacific Blue) IgG1a 5 558121 BD Biosciences 1, 2
CD33 (PerCP-Cy5.5) IgG1 20 333146 BD Biosciences 1, 2
CD7 (FITC) IgG1 20 555360 BD Pharmingen 1, 2
CD4 (Alexa 700) IgG1 5 557922 BD Pharmingen 1, 2
CD25 (PE-Cy 7) IgG1 5 557741 BD Pharmingen 1, 2
CD3 (Biotin)
c
IgG2a 20 555338 BD Pharmingen 1
CD8 (Alexa 647) IgG1 5 557708 BD Pharmingen 1
CD19 (Biotin)
c
IgG1 20 555411 BD Pharmingen 2
CD56 (APC) IgG1 20 555518 BD Pharmingen 2
CD34 (PE Cy7) IgG1 5 348811 BD Sciences 3
CD133 (APC) IgG2b 5 130-090-854 Miltenyi Biotec 3
CD38 (Biotin)
c
IgG1 5 ab30418 AbCam 3
CD90 (FITC) IgG1 5 555595 BD Biosciences 3
CD117 (unconjugated)
b
IgG1 5 313202 Bio legend 3
CD41 (unconjugated)
a
IgG1 5 ab15021 AbCam 3
Lineage 1 (FITC) IgG1/2b 20 340546 BD Biosciences 4
CD123 (PerCP-Cy5) IgG2a 20 558714 BD Pharmingen 4
CD11c (PE) IgG1 5 555392 BD Pharmingen 4
HLA-DR (Alexa 700) IgG2a 5 307626 Bio legend 4
HLA-DQ (Biotin)
c
IgG2a 5 ab24265 AbCam 4
HLA-ABC (Pacific Blue) IgG2a 5 311418 Bio legend 4
HLA-G (unconjugated)
a
IgG1 5 ab7758 AbCam 4
CD16 (PE-Cy7) IgG1 5 335823 BD Biosciences 4
Unconjugated antibodies were labelled using a Zenon labelling complex from Invitrogen; Alexa405 z25313
b
or Alexa430 z25301
a
. The
biotinylated antibodies were labelled using Quantum dots 605, also from Invitrogen, conjugated to streptavidinc
.
04534. USA), making a final volume of 4 ml. Samples
were plated in triplicate in small, gridded Petri dishes
(Nunc brand, VWR 734-2114 Westchester PA, USA) 1ml
per plate. The plates were incubated at 37o
C with 5% CO2
for 14 days. After incubation the number of granulo-
cyte-macrophage colonies and other lineages present were
assessed. An average colony number from the three test
plates was then calculated to give colony numbers per 105
cells plated.
Flow cytometric analysis: Flow cytometric analysis was
carried out using a Becton Dickinson FACS Caliber ma-
chine. The samples were prepared as follows; 100μl of
blood was added to each tube and 50μl of antibody cock-
tail (For antibody, fluorochromes and supplier details see
Table 1). The tubes were then incubated at room temper-
ature, in the dark for 20 min. The cells were then lysed
and washed on the BD FACS Lyse/Wash Assistant, after
which the samples were run on the flow cytometer.
Cryopreservation of umbilical cord blood: When the
UCB was ready for cryopreservation it was cooled to 4o
C
before being transferred to a cryostore bag (Quest Biome-
dical CS250n. Solihull, UK.), where dimethyl sulphoxide
(DMSO) (Origen 210002, Austin TX, USA) mixed with
Dextran 40 (Baxter Healthcare B5043), also cooled to 4o
C,
was added at a concentration of 10%. The sample was cry-
opreserved in a controlled rate freezer (Planar Kryo 560-
16. Sunbury-on-Thames, UK) using the following proto-
col: Start temperature=4o
C, step I=hold @ 4
o
C for 10
min, step II=−2o
C/min to −5
o
C, step III=−1
o
C/min to
−40o
C, step IV=−5
o
C/min to −100
o
C. The sample was
then transferred to the gaseous phase of a liquid nitrogen
Dewar. Samples were frozen for a minimum 14 days be-
fore thawing.
Thawing of umbilical cord blood: This protocol was
based on Rubinstein’s method (14). After thawing, the
sample was then resuspended in Dextran/Albumin to its
original post processing volume.
Statistical analysis
Statistical analysis was carried out using the statistical
software programme, Minitab (Version 15, 2006, Univer-
sity of Pennsylvania, USA). The data was analysed using
5. 36 International Journal of Stem Cells 2010;3:32-45
Fig. 1. Sepax gives an increased recovery of nucleated cells than all other methods. (A) Sepax offers the highest recovery of CD45+ cells
at 75.79% (SD±18.58) compared to PrepaCyte CB at 72.03% (SD±8.48), Plasma depletion at 71.38% (SD±29.35), Hetastarch at 44.94%
(SD±20.06), and Density Gradient at 19.13% (SD±10.88). Sepax was only significantly more efficient than Hetastarch and density gradient
methods (p<0.005). (B) After exclusion of granulocytes recovery with Sepax is still the greatest at 78.8% (SD±21.36) compared to other
methods. These results were determined using differential cell counts coupled with flow cytometric analysis of CD45+ cells, viability of
CD45+ cells. Exclusion of granulocytes was also determined in this way to give a more accurate picture of cell recovery. Fluorochromes
used were PE Cy5 for CD45+ cells, PE for CD14 +/− cells and 7AAD for viability.
non parametric, two-tailed Mann-Whitney U-testing to de-
termine significance. A p<0.05 was considered to be stat-
istically significant.
Results
For this study, 80 UCB units were processed. Of those
samples, 46% were from female infants (n=37) and 54%
were from male infants (n=43). The birth weights of the
infants involved ranged from 2.41 kg to 4.42 kg with an
average of 3.52 kg (SD±0.38). The volume of blood col-
lected ranged from 46.6 ml to 194.3 ml with an average
volume of 98.72 ml (SD±31.56).
Recovery of nucleated cell fraction
When looking at TNC recovery, Sepax gives the highest
recovery at 75.79% (SD±18.58). This is significantly grea-
ter than all other methods tested (p<0.05) apart from
PrepaCyte-CB (p=0.90) (Fig. 1A). After exclusion of gran-
ulocytes recovery with Sepax is still the greatest at 78.8%
(SD±21.36) compared to other methods, however data is
only significant when compared to density gradient sepa-
ration where recovery is 40.28% (SD±20.97) (p<0.01).
These results were determined using differential cell counts
coupled with flow cytometric analysis of CD45+ cells, vi-
ability of CD45+ cells. Exclusion of granulocytes was also
determined in this way to give a more accurate picture
of cell recovery. Fluorochromes used were PE Cy5 for
CD45+ cells, PE for CD14 +/− cells and 7AAD for
viability. We also examined the effect initial cord blood
volume had on recovery of nucleated cells. Hetastarch,
Sepax (not significant) and plasma depletion processing
exhibited a negative correlation; as the volume of the cord
blood increased, the recovery of nucleated cells decreased
(p<0.05) (Fig. 6A, D, E). However when processed with
PrepaCyte-CB there was no significant correlation and re-
covery remained constant regardless of volume (Fig. 6B).
Density gradient processing became more efficient as
UCB volume increased (p<0.01) (Fig. 6C).
Recovery of haematopoietic stem cells
We looked at three different populations of HSC as
previously described; early stage (CD34−/CD133+),
mid stage (CD34+/CD133+) and late stage (CD34+/
CD133−) (13). When looking at recovery of late stage
haematopoietic stem cells results show that PrepaCyte-CB
gives the greatest recovery at 74.47% (SD±8.89). Although
this result is higher than all other methods it is only sig-
nificantly more effective than Hetastarch, whose recovery
was low at 56.48% (SD±21.99), providing an average in-
crease of 17.99% (p=0.024) (Fig. 2A). Again we looked at
the effect initial cord blood volume had on recovery of
CD34+ cells. As was shown with nucleated cell recovery,
Hetastarch, Sepax and plasma depletion processing ex-
6. Christina Basford, et al: The Cord Blood Separation League Table 37
Fig. 2. PrepaCyte-CB provides the most efficient recovery of haematopoietic stem cells than other methods tested. (A) Recovery of late
stage haematopoietic (CD34+/CD133−) stem cells was greater with PrepaCyte-CB at 74.41% (SD±8.89) than Sepax; 73.91% (SD±16.86),
plasma depletion; 73.88% (SD±22.26) Hetastarch; 56.48% (SD±21.99) and density gradient separation methods 22.83% (SD±18.56).
However, results are only significant when compared to HES (p=0.05). (B) Recovery of mid stage haematopoietic (CD34+/CD133+) stem
cells stem cells was greater with PrepaCyte-CB at 65.83% (SD±14.22) than Hetastarch; 64.21% (SD±17.89), Sepax; 60.98% (SD±14.90),
plasma depletion; 54.30% (SD±26.79) and density gradient separation methods 27.43% (SD±13.37). However, results are only significant
when compared to density gradient (p=<0.01). (C) Recovery of early stage haematopoietic (CD34−/CD133+) stem cells was greater
with PrepaCyte-CB at 62.59% (SD±16.23) than plasma depletion; 60.10% (SD±28.5), Sepax; 48.50% (SD±15.06) Hetastarch; 48.40%
(SD±26.2) and density gradient separation methods 13.06% (SD±9.36). However, results are only significant when compared to density
gradient (p=<0.01). CD34+ cell numbers and viability were calculated using flow cytometric analysis of CD34 antibody and 7AAD uptake.
Flourochromes used were FITC for CD45+ cells, PE for CD34+ cells and 7AAD for viability.
hibited a negative correlation; as the volume of the cord
blood increased, the recovery of CD34+ cells decreased
(p<0.05) (Fig. 6F, I, J). However when processing with
PrepaCyte-CB there was no significant correlation and re-
covery remained constant regardless of volume (Fig. 6G).
Density gradient processing became more efficient as
UCB volume increased (p<0.01) (Fig. 6H).
Next we looked at recovery of mid stage HSC.
PrepaCyte-CB gave the highest recovery at 65.83%
(SD±14.22); this was greater than all the others methods
evaluated but only significantly higher than density gra-
dient processing which gave a recovery of 27.43%
(SD±13.37) (Fig. 2B).
Finally when looking at recovery of early stage, it was
7. 38 International Journal of Stem Cells 2010;3:32-45
Fig. 3. PrepaCyte-CB gives the greatest recovery of T Cells (CD45+/CD3+) and B Cells (CD45+/CD19+). (A) Recovery of T Cells (CD45+
/CD3+) was greater with PrepaCyte-CB at 71.84% (SD±18.06) than plasma depletion; 65.54% (SD±35.0), Sepax; 65.26% (SD±35.19)
Hetastarch; 56.85% (SD±23.79) and density gradient separation methods 36.62% (SD±26.32). (B) Recovery of B Cells (CD45+/CD19+)
was greater with PrepaCyte-CB at 75.68% (SD±24.0) than Sepax; 71.75% (SD±16.84), plasma depletion; 71.74% (SD±8.66) Hetastarch;
46.47% (SD±25.69) and density gradient separation methods 19.13% (SD±10.88). However, results are only significant when compared
to HES (p=<0.05).
Fig. 4. Density gradient separation is the most effective method for volume reduction by removal of either; (A) Red blood cells and (B)
Haemoglobin. (A) Compared to whole blood, which has an average of 2.9×106
cell/ml (SD±0.75), post processing, density gradient is
the most effective method for the removal of red blood cells, it leaves 0.03×106
cell/ml (SD±0.02), PrepaCyte-CB leaves 0.24×106
cell/ml
(SD±0.08), Hetastarch leaves 1.93×10
6
cell/ml (SD±0.22), plasma depletion leaves 2.01×106
cell/ml (SD±1.53) and Sepax leaves
3.33×106
cell/ml (SD±0.95), all results are significant (p=<0.05). Post thaw density gradient is again the most effective method for the
removal of red blood cells, it leaves 0.02×106
cell/ml (SD±0.01), PrepaCyte-CB leaves 0.11×10
6
cell/ml (SD±0.10), Hetastarch leaves
0.22×10
6
cell/ml (SD±0.16), plasma depletion leaves 0.54×106
cell/ml (SD±0.22) and Sepax leaves 2.01×106
cell/ml (SD±0.94), all
results are significant (p=<0.05). (B) Compared to whole blood, which has an average of 10.58 g/dl (SD±4.11), post processing, density
gradient is the most effective method for the removal of haemoglobin, it leaves 0.17 g/dl (SD±0.12), PrepaCyte-CB leaves 0.68 g/dl
(SD±0.31), Hetastarch leaves 6.94 g/dl (SD±4.74), plasma depletion leaves 7.23 g/dl (SD±5.56) and Sepax leaves 13.07 g/dl (SD±7.03),
all results are significant (p=<0.05). Post thaw PrepaCyte-CB is the most effective method for the removal of haemoglobin, it leaves 0.10
g/dl (SD±0.04), density gradient leaves 0.13 g/dl (SD±0.05), Hetastarch leaves 0.46 g/dl (SD±0.29), plasma depletion leaves 1.94 g/dl
(SD±1.05) and Sepax leaves 7.57 g/dl (SD±3.70), all results are significant (p=<0.05).
8. Christina Basford, et al: The Cord Blood Separation League Table 39
again PrepaCyte-CB which gave the greatest recovery
62.59% (SD±16.23). This is significantly higher than sep-
aration using Sepax and density gradient (p<0.05) (Fig.
2C).
Recovery of other cells types within UCB
When looking at T cells (CD45+/CD3+) recovery,
PrepaCyte-CB gives the highest recovery at 71.84% (SD±
18.06). This is significantly higher than Hetastarch (p<
0.05) (Fig. 3A). For B cells (CD45+/CD19+) recovery,
plasma depletion gives the highest recovery at 75.68%
(SD±24.0). This is significantly higher than PrepaCyte-
CB and Hetastarch (p<0.05) (Fig. 3B).
Volume reduction
For red blood cell removal, density gradient separation
was the most effective method. The average number of red
blood cells per ml of whole blood was 2.92×106
cells (SD
±0.75). After processing with density gradient methods
the number of red blood cells was reduced to 0.03×106
cells/ml of blood (SD±0.02). This reduction was signifi-
cantly greater than with PrepaCyte-CB where RBC num-
bers were on average 0.24×106
cells/ml (SD±0.08), a 12
fold decrease (p=0.00005), with plasma depletion where
RBC numbers were on average 2.01×106
cells/ml (SD±
1.53), a 0.6 fold decrease (p=0.00003), with HES where
RBC numbers were 1.93 106
/ml (SD±1.32) a 1.5 fold de-
crease (p=0.005) and with Sepax where RBC numbers
were 3.33×106
cells/ml (SD±1.53), a 0.9 fold decrease
(p=0.000005) (Fig. 4A). We also examined whether initial
collected cord blood volume had an effect on RBC deple-
tion but our data revealed no correlation. We also exam-
ined whether initial collected volume of UCB had an im-
pact on RBC depletion but no correlation was observed.
For removal of haemoglobin density gradient was the
most efficient. Whole blood had an average of 10.58 g/dl
of haemoglobin (SD±4.11), after density gradient process-
ing this was reduced to 0.17 g/dl (SD±0.12). Density gra-
dient was 76 fold more efficient at removing haemoglo-
bin than Sepax (p=0.000009) where post processing levels
were 13.07 g/dl (SD±5.56), it was 43 fold more efficient
at Haemoglobin removal than plasma depletion (p=0.0008)
where post processing levels were 7.23 g/dl (SD±7.03), it
was 41 fold more efficient than HES (p=0.006) where post
processing levels were 6.94 g/dl (SD±4.74), and it was 4
fold more efficient than PrepaCyte-CB (p=0.002) where
post processing levels were 0.68 g/dl (SD±0.31) (Fig. 4B).
Clonogenic potential
When processing with PrepaCyte-CB, UCB units are
left with the highest clonogenic potential compared to the
other methods; 13.27 CFU/108
cells (SD±3.33). This is
significantly higher than with density gradient separation;
where clonogenic potential was 5.54 CFU/108 cells (SD±
5.36), 2.3 fold greater (p=0.0002), plasma depletion; where
clonogenic potential was 8.59 CFU/108
cells (SD±1.65),
1.6 fold greater (p=0.00003), and Sepax; where clonogenic
potential was 11.10 CFU/108 cells (SD±2.89), 1.2 fold
greater (p=0.05). Although PrepaCyte-CB also provided a
higher clonogenic potential than Hetastarch these data
was not significant (p=0.17). After thaw results are as fol-
lows: PrepaCyte-CB processing left UCB units 9.23 CFU/
108
cells (SD±2.31), the highest clonogenic potential com-
pared to the other methods. This is significantly higher
than with Sepax separation; where clonogenic potential
was 6.08 CFU/108
cells (SD±2.15), 1.5 fold greater (p=
0.0003), plasma depletion; where clonogenic potential was
5.70 CFU/108
cells (SD±1.06), 1.6 fold greater (p=0.000003),
Hetastarch; where clonogenic potential was 3.94 CFU/108
cells (SD±1.44), 6.4 fold greater (p=0.000009), and densi-
ty gradient separation: where clonogenic potential was 1.33
CFU/108
cells (SD±2.31), 6.9 fold greater (p=0.000009)
(Fig. 5).
Discussion
Recently there has been an explosion of clinical applica-
tions for UCB. This escalation leads to an increased need
for optimising separation, whether to reduce volume to
make storage more efficient or to increase stem cell yield
for transplant. UCB potential is well documented, with
enough clinical value to warrant further investigation.
Since the advent of regular UCB transplantation in the
1990s, and the potential of the units increased, it was nec-
essary to increase banking to meet the demand. It may
also be linked to limitations of BM registries. The process
of finding a donor can be long; potential donors may
change address without notifying their registry; ethnicity
can also be an issue with ethnic minorities often strug-
gling to find a suitable donor. UCB can help towards this
problem as it is easier to store samples from donors of
many ethnicities (21). UCB is an accessible option, with
samples already stored. However this increase is not in
line with sample demand, and the contrast between these
two resources is significant. There are currently 11 million
patrons registered worldwide for BM donation, only an es-
timated 300,000 UCB units are publically banked. Some
of these banks are struggling financially so it is of critical
importance to find the most effective and economical
processing, and storage methods (22). Many factors affect
9. 40 International Journal of Stem Cells 2010;3:32-45
Fig. 5. PrepaCyte-CB processing leaves samples with the highest
clonogenic potential both post processing and post thaw. Whole
blood has on average 10.22 CFU per 10
8
cells (SD±3.80). Post
processing PrepaCyte-CB leaves the greatest number of CFU at
13.28 per 10
8
cells (SD±5.36) compared to Hetastarch which
leaves 11.53 per 10
8
cells (SD±1.65), Sepax which leaves 11.10
per 10
8
cells (SD±2.89), plasma depletion which leaves 8.59 per
108
cells (SD±4.74), and density gradient which leaves 5.55 per
10
8
cells (SD±3.36). Samples processed with PrepaCyte-CB have
a significantly greater clonogenic potential than all other methods
except Hetastarch (p=<0.01). Post thaw PrepaCyte-CB again
leaves the greatest number of CFU at 9.23 per 10
8
cells (SD±2.31)
compared to Hetastarch which leaves 3.94 per 10
8
cells
(SD±1.06), Sepax which leaves 6.08 per 108
cells (SD±2.15), plas-
ma depletion which leaves 5.70 per 108
cells (SD±1.44), and den-
sity gradient which leaves 1.33 per 108
cells (SD±2.33). Samples
processed with PrepaCyte-CB have a significantly greater clono-
genic potential than all other methods (p=<0.01).
the success of processing, not least the physicality of nor-
mal birthing (13), so it remains an important priority to
find the best possible methods of separation, so that even
smaller units may prove clinically viable.
Our results show that Sepax depletion gives a higher re-
covery of nucleated cells, crucial for successful engraft-
ment. However, recovery using Sepax is reduced as the
size of unit processed increases. Hetastarch, density gra-
dient and plasma depletion separation were also affected
in this way, but PrepaCyte-CB processing was not affected
by the initial volume of the collected unit, and recovery
of both TNC and CD34+ progenitor cells was as efficient
with smaller volumes as it was with larger units. Density
gradient separation shows a reverse correlation: as UCB
volume increase, so does recovery. Although interesting,
it does not fairly compare to the other methods as the
maximum volume processed with density gradient was 90
ml and all other methods were routinely tested with units
of over 100 ml. The National Cord Blood Program at the
New York Blood Center suggests an average TNC count
for a UCB transplant should be a minimum of 2×107
cells
per kg of body weight (23). Recent findings show that it
is now as successful to transplant an adult patient with
multiple UCB units as it is to transplant a child with a
high dose single unit (24). HLA is also important when
considering units for transplantation, ideally it would be
a 6/6 match but, for 5/6 or less an increase TNC is essen-
tial to promote engraftment (25). From the results of this
study we believe that Sepax, offers the best recovery of
TNC, with PrepaCyte-CB and plasma depletion close
behind. However, it should also be noted that the content
of plasma depleted product is a concern for transplant in
conditions other than haematopoietic reconstitution. Fur-
ther, in our experiments it was not possible to differ-
entiate plasma depleted product, even after further proc-
essing into hepatic lineages, despite this being possible for
other processing systems such as PrepaCyte-CB, Sepax
and density gradient (Hetastarch was not tested).
We next focused on recovery of haematopoietic stem
cells of three different developmental stages; our study
shows that PrepaCyte-CB offers the best methodology for
optimum HSC numbers from all three stages, although
again it is worth mentioning that Sepax recovery for both
CD34+ and TNC is diminished as the volume of the
UCB increases. When the number of HLA mismatches is
greater, then CD34+ count is more critical (26). Suggest-
ed minimum numbers are 1.7×105
per kg (27). The bene-
fits of Sepax are that it is a fully automated system which
allows for the mass processing of samples, which is suit-
able for larger cord blood banks.
Additionally we examined recovery of some immune
cells: T and B cells. T cells which are involved in cell
mediated immunity. They can be stimulated by antigens
and have dual function, for example; T helper cells and
cytotoxic T cells, or can act as regulatory T cells, which
are involved in immunological tolerance by suppressing T
cell mediated immunity at the end of an immune response
and by halting auto-immune reactions (28). Also B cells,
which are involved in the humoral immune response.
These cells are activated in the presence of foreign antigen
to make antibodies. However, this is a complex process
and assistance is required from T helper cells (29). During
this study we found superior recovery of CD45+/CD3+
T lymphocytes with PrepaCyte CB which also gave the
best results for CD10+ B cells. Not much is known about
whether these cells play a role in engraftment in humans
but some mice models have shown that increased numbers
10. Christina Basford, et al: The Cord Blood Separation League Table 41
Fig. 6. Initial cord blood volume negatively correlates with recovery of CD45+ and CD34+ haematopoietic progenitor cells when process-
ing with Hetastarch, Plasma Depletion and Sepax. For CD45+ cell recovery: (A) Hetastarch processing negatively correlates with initial
volume (p<0.01). (B) Initial volume has no significant impact on recovery of TNC when processing with PrepaCyte-CB (p>0.05). (C) Density
gradient processing has a positive correlation with an increase in collected volume (p<0.01). (D) Plasma Depletion separation negatively
correlates with initial UCBV volume (p<0.01). (E) Sepax processing also negatively correlates with initial UCB volume but not significantly
(p>0.05). For CD34+ recovery results are the same. (F) Hetastarch (p<0.01). (G) PrepaCyte-CB (p>0.05). (H) Density gradient (p<0.05).
(I) Plasma Depletion (p<0.01). (J) Sepax processing negatively correlates with initial UCB volume (p<0.05).
11. 42 International Journal of Stem Cells 2010;3:32-45
Fig. 6. Continued.
of T cells transplanted can increase bone marrow recon-
stitution and therefore haematopoiesis and also eliminate
residual leukemic disease in the transplanted mice. How-
ever, it is important to get the fullest picture of the unit
quality that we can.
When looking at the volume reduction of the physical
size of the unit, it would seem that plasma depletion
would be of particular benefit, as a smaller volume re-
duces the space needed for storage and also means less
DMSO is added to the sample in preparation for cry-
opreservation (20). This means that it could even save the
need to wash samples before infusion (for haematopoietic
transplant only) as it has been previously shown that TNC
recovery after cryopreservation is greater without a wash
step (30). However, if you measure volume reduction as
the ability to remove RBC and Haemoglobin; then it is
actually a simple and economic density gradient separa-
tion which is the most efficient. Removal of RBC is also
a well documented way to reduce the bulk of UCB units
(14). Ficoll-paque was the most successful at removing
both RBC and haemoglobin. Banking of UCB units is a
fiercely discussed topic at the moment with much focus
on the debate of private versus public banks (31) so it is
important to get the methods right, whichever style of
banking the cord blood unit is destined for.
Finally we looked at clonogenic potential of the UCB
units which was measured by the CFU assay. Traditionally,
this is the most important test, since it gives the best pos-
sible readout for potential of the cord blood to be useful
if used therapeutically. PrepaCyte-CB performed best in
this test, not only post processing but also after cryopre-
servation and the subsequent thawing. The significance of
post thaw CFU is critical for future therapeutic uses of
UCB units as it is necessary to know that they will still
be able to engraft after storage (32). This could be because
PrepaCyte-CB is the second most efficient method for re-
12. Christina Basford, et al: The Cord Blood Separation League Table 43
Table 2. Processing league table
Method
Performance position & frequency
Points
1st
a
2ndb
3rdc
1 PrepaCyte-CB 4 4 1 21
2 Sepax 2 3 2 14
3 Plasma Depletion 2 1 4 12
4 Density gradient 2 1 0 8
5 Hetastarch 0 1 3 5
a
1st place is worth 3 points, b
2nd place is worth 2 points, c
3rd
place is worth 1 point (9 methods of analysis).
moving RBC. A reduction in RBC numbers has previously
been shown to have a advantageous effect on CFU (16,
33). The fact that recovery of CD34+ cells did not fit well
with the clonogenic potential also fits with our previously
reported theories that many CD34+ populations are rela-
tively late for haematopoietic engraftment (such as CD34
+/45+) and that cells earlier than CD34 exist in cord
blood units. This is an important point to consider when
carrying out cord blood banking because it means that
those banks, who only discuss CD34+ cells and/or TNC,
are not representing the whole picture of cord blood.
We also analysed the cost of processing a single unit
using each of the five methods. Costs were worked out by
pricing the general plastic ware, the separation reagents
and any special, non-usual laboratory equipment required.
For some of methods where special kits were used both
the list price option and the best available price (for exam-
ple using a large number of kits of a yearly period) were
taken into consideration. Plasma depletion followed by den-
sity gradient was the cheapest methods analysed. Sepax
was the most expensive.
During the analysis no one method consistently came
out best; so a league table was devised. Each method was
rated for all nine analysis methods (as discussed above)
and was awarded 3 points for 1st
place, 2 points for 2
nd
place and finally 1 point for 3rd
place (Table 2). Plasma
Depletion has proved to be a cost effective, rapid and effi-
cient method for volume reduction of UCB. Sepax was
proven to be the most efficient method for TNC recovery.
Only Hetastarch didn’t perform best in any of the tests.
This system showed that PrepaCyte-CB is consistently the
best performer over the whole range of analysis. Prepa-
Cyte-CB has other benefits as it is a closed system, re-
ducing the risk of contamination. Not only is this factor
useful to the USA and the rest of the developed world but
may serve to help UCB storage a more viable option in
less economically developed parts of the world where ac-
cess to a clean room may not be available. The Prepa-
Cyte-CB system does not require expensive laboratory
equipment and this is a significant issue in the developing
countries where transplants are rarely carried out due to
lack of necessary equipment. It is worth noting that UCB
transplants are currently unavailable to large sections of
the second and third worlds.
In conclusion we can say that, depending on your pri-
mary use for UCB, different processing methods may be
more applicable than others, but that while ‘futureproof-
ing’ is not easy in medical sciences, the choice of process-
ing method must be carefully addressed so that patients
have cord blood units available in a suitable form to treat
their disease.
Acknowledgments
With thanks to the staff at Newcastle’s Royal Victoria
Infirmary Woman’s Directorate and Maternity Unit. We
are grateful to One North East regional development agen-
cy of the British Government for their financial support
to our laboratories.
Potential Conflict of Interest
The authors have no conflicting financial interest.
References
1. Gluckman E, Rocha V. Cord blood transplantation: state of
the art. Haematologica 2009;94:451-454
2. Gluckman E, Broxmeyer HA, Auerbach AD, Friedman HS,
Douglas GW, Devergie A, Esperou H, Thierry D, Socie G,
Lehn P. Hematopoietic reconstitution in a patient with
Fanconi's anemia by means of umbilical-cord blood from
an HLA-identical sibling. N Engl J Med 1989;321:1174-1178
3. Escolar ML, Poe MD, Provenzale JM, Richards KC, Allison
J, Wood S, Wenger DA, Pietryga D, Wall D, Champagne
M, Morse R, Krivit W, Kurtzberg J. Transplantation of um-
bilical-cord blood in babies with infantile Krabbe's disease.
N Engl J Med 2005;352:2069-2081
4. Slatter MA, Gennery AR. Umbilical cord stem cell trans-
plantation for primary immunodeficiencies. Expert Opin
Biol Ther 2006;6:555-565
5. Davey S, Armitage S, Rocha V, Garnier F, Brown J, Brown
CJ, Warwick R, Fehily D, Watt S, Gluckman E, Vora A,
Contreras M, Navarrete CV. The London Cord Blood Bank:
analysis of banking and transplantation outcome. Br J
Haematol 2004;125:358-365
6. Fasouliotis SJ, Schenker JG. Human umbilical cord blood
banking and transplantation: a state of the art. Eur J Obstet
Gynecol Reprod Biol 2000;90:13-25
7. Grewal SS, Barker JN, Davies SM, Wagner JE. Unrelated
donor hematopoietic cell transplantation: marrow or um-
bilical cord blood? Blood 2003;101:4233-4244
13. 44 International Journal of Stem Cells 2010;3:32-45
8. Kekarainen T, Mannelin S, Laine J, Jaatinen T. Optimiza-
tion of immunomagnetic separation for cord blood-derived
hematopoietic stem cells. BMC Cell Biol 2006;730
9. Broxmeyer HE, Gluckman E, Auerbach A, Douglas GW,
Friedman H, Cooper S, Hangoc G, Kurtzberg J, Bard J,
Boyse EA. Human umbilical cord blood: a clinically useful
source of transplantable hematopoietic stem/progenitor
cells. Int J Cell Cloning 1990;8 Suppl 1:76-89
10. Tondreau T, Meuleman N, Delforge A, Dejeneffe M, Leroy
R, Massy M, Mortier C, Bron D, Lagneaux L. Mesenchymal
stem cells derived from CD133-positive cells in mobilized
peripheral blood and cord blood: proliferation, Oct4 ex-
pression, and plasticity. Stem Cells 2005;23:1105-1112
11. Denner L, Bodenburg Y, Zhao JG, Howe M, Cappo J, Tilton
RG, Copland JA, Forraz N, McGuckin C, Urban R.
Directed engineering of umbilical cord blood stem cells to
produce C-peptide and insulin. Cell Prolif 2007;40:367-380
12. McGuckin C, Forraz N, Baradez MO, Basford C, Dickinson
AM, Navran S, Hartgerink JD. Embryonic-like stem cells
from umbilical cord blood and potential for neural mo-
deling. Acta Neurobiol Exp (Wars) 2006;66:321-329
13. McGuckin CP, Basford C, Hanger K, Habibollah S, Forraz
N. Cord blood revelations-The importance of being a first
born girl, big, on time and to a young mother! Early Hum
Dev 2007;83:733-741
14. Rubinstein P, Dobrila L, Rosenfield RE, Adamson JW,
Migliaccio G, Migliaccio AR, Taylor PE, Stevens CE.
Processing and cryopreservation of placental/umbilical cord
blood for unrelated bone marrow reconstitution. Proc Natl
Acad Sci USA 1995;92:10119-10122
15. Vannier JP, Monconduit M, Piguet H. Comparison between
2 density gradients for separation of CFU. Biomedicine
1980;33:236-239
16. Solves P, Mirabet V, Planelles D, Blasco I, Perales A,
Carbonell-Uberos F, Soler MA, Roig R. Red blood cell de-
pletion with a semiautomated system or hydroxyethyl starch
sedimentation for routine cord blood banking: a com-
parative study. Transfusion 2005;45:867-873
17. Chow R, Nademanee A, Rosenthal J, Karanes C, Jaing TH,
Graham ML, Tsukahara E, Wang B, Gjertson D, Tan P,
Forman S, Petz LD. Analysis of hematopoietic cell trans-
plants using plasma-depleted cord blood products that are
not red blood cell reduced. Biol Blood Marrow Transplant
2007;13:1346-1357
18. Ademokun JA, Chapman C, Dunn J, Lander D, Mair K,
Proctor SJ, Dickinson AM. Umbilical cord blood collection
and separation for haematopoietic progenitor cell banking.
Bone Marrow Transplant 1997;19:1023-1028
19. Berger MJ, Adams SD, Tigges BM, Sprague SL, Wang XJ,
Collins DP, McKenna DH. Differentiation of umbilical
cord blood-derived multilineage progenitor cells into respi-
ratory epithelial cells. Cytotherapy 2006;8:480-487
20. Lapierre V, Pellegrini N, Bardey I, Malugani C, Saas P,
Garnache F, Racadot E, Schillinger F, Maddens S. Cord
blood volume reduction using an automated system (Sepax)
vs. a semi-automated system (Optipress II) and a manual
method (hydroxyethyl starch sedimentation) for routine cord
blood banking: a comparative study. Cytotherapy 2007;9:
165-169
21. Meyer-Monard S, Passweg J, Troeger C, Eberhard HP,
Roosnek E, de Faveri GN, Chalandon Y, Rovo A, Kindler
V, Irion O, Holzgreve W, Gratwohl A, Müller C, Tichelli
A, Tiercy JM. Cord blood banks collect units with different
HLA alleles and haplotypes to volunteer donor banks: a
comparative report from Swiss Blood stem cells. Bone Mar-
row Transplant 2009;43:771-778
22. McGuckin CF, Forraz N. Umbilical cord blood stem cells--
an ethical source for regenerative medicine. Med Law 2008;
27:147-165
23. Wagner JE, Barker JN, DeFor TE, Baker KS, Blazar BR,
Eide C, Goldman A, Kersey J, Krivit W, MacMillan ML,
Orchard PJ, Peters C, Weisdorf DJ, Ramsay NK, Davies
SM. Transplantation of unrelated donor umbilical cord
blood in 102 patients with malignant and nonmalignant dis-
eases: influence of CD34 cell dose and HLA disparity on
treatment-related mortality and survival. Blood 2002;100:
1611-1168
24. Sauter C, Barker JN. Unrelated donor umbilical cord blood
transplantation for the treatment of hematologic malig-
nancies. Curr Opin Hematol 2008;15:568-575
25. Lister J, Gryn JF, McQueen KL, Harris DT, Rossetti JM,
Shadduck RK. Multiple unit HLA-unmatched sex-misma-
tched umbilical cord blood transplantation for advanced
hematological malignancy. Stem Cells Dev 2007;16:177-186
26. Rodrigues CA, Sanz G, Brunstein CG, Sanz J, Wagner JE,
Renaud M, de Lima M, Cairo MS, Fürst S, Rio B, Dalley
C, Carreras E, Harousseau JL, Mohty M, Taveira D, Dreger
P, Sureda A, Gluckman E, Rocha V. Analysis of risk factors
for outcomes after unrelated cord blood transplantation in
adults with lymphoid malignancies: a study by the euro-
cord-netcord and lymphoma working party of the european
group for blood and marrow transplantation. J Clin Oncol
2009;27:256-263
27. Van Haute I, Lootens N, De Buck K, Verdegen L, Vander
Steene V, Desmet S, Craeye D, Vandekerckhove B. Selecting
cord blood units for storage by CD34+ cell counts. Trans-
fusion 2005;45:455-457
28. Adkins B. T-cell function in newborn mice and humans.
Immunol Today 1999;20:330-335
29. Li J, Barreda DR, Zhang YA, Boshra H, Gelman AE,
Lapatra S, Tort L, Sunyer JO. B lymphocytes from early
vertebrates have potent phagocytic and microbicidal abili-
ties. Nat Immunol 2006;7:1116-1124
30. Laroche V, McKenna DH, Moroff G, Schierman T, Kadidlo
D, McCullough J. Cell loss and recovery in umbilical cord
blood processing: a comparison of postthaw and postwash
samples. Transfusion 2005;45:1909-1916
31. Committee on Obstetric Practice; Committee on Genetics.
ACOG committee opinion number 399, February 2008: um-
bilical cord blood banking. Obstet Gynecol 2008;111:475-477
32. Yoo KH, Lee SH, Kim HJ, Sung KW, Jung HL, Cho EJ,
Park HK, Kim HA, Koo HH. The impact of post-thaw col-
14. Christina Basford, et al: The Cord Blood Separation League Table 45
ony-forming units-granulocyte/macrophage on engraftment
following unrelated cord blood transplantation in pediatric
recipients. Bone Marrow Transplant 2007;39:515-521
33. de Kreuk AM, Zevenbergen A, van Oostveen JW, Schuur-
huis GJ, Huijgens PC, Jonkhoff AR. A single-step col-
ony-forming unit assay for unseparated mobilized periph-
eral blood, cord blood, and bone marrow. J Hematother
Stem Cell Res 2001;10:795-806
34. Samuel GN, Kerridge IH, O'Brien TA. Umbilical cord
blood banking: public good or private benefit? Med J Aust
2008;188:533-535.