This document describes the facilities and research at ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione), a transplant hospital and research foundation in Palermo, Italy. The document outlines ISMETT's laboratories for regenerative medicine, immunobiology, and biomedical technologies. It also summarizes some of ISMETT's research areas like adoptive immunotherapies, fetal hepatocyte transplantation, pancreatic islet transplantation, and characterization of stem/progenitor cells. Services mentioned include development of adoptive immunotherapies, stem cell production in GMP facilities, and clinical trials.
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
HYBRIDOMA TECHNOLOGY IT IS DEFINED AS THE PROCESS WERE THERE IS A FUSION OF SPLLEN CELL AND MYELOMA CELLS IN THE PRESENCE OF POLYETHYLENE GLYCOL OR SENDAI VIRUS AND LEADS TO THE PRODUCTION OF MONOCLONL ANTIBODY.
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
HYBRIDOMA TECHNOLOGY IT IS DEFINED AS THE PROCESS WERE THERE IS A FUSION OF SPLLEN CELL AND MYELOMA CELLS IN THE PRESENCE OF POLYETHYLENE GLYCOL OR SENDAI VIRUS AND LEADS TO THE PRODUCTION OF MONOCLONL ANTIBODY.
Sanja Selak of Intercell AG, Vienna, Austria, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Intercell develops vaccines for the prevention and treatment of infectious diseases
Gene Olinger, USAMRIID, Fort Detrick USA, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Protective Immune Reponses to Ebola Virus
Paludisme grave : pourquoi doit-on développer des modèles in vitro sur le terrain ? - Conférence de la 8e édition du Cours international « Atelier Paludisme » - RAZAKANDRAINIBE Romy - Madagascar - romy@pasteur.mg
ProImmune Antigen Characterization Summit Paul Mossamandacturner
Paul Moss, School of Cancer Sciences, Birmingham UK, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Cytomegalovirus and Cancer-specific Immunity
Non-Viral Φc31 Integrase Mediated In Vivo Gene Delivery to the Adult Murine Kidney by Daniel C Chung, Matthew C Canver, Xiaofeng Zuo and Jean Bennett and Joshua H Lipschutz* in Experimental Techniques in Urology & Nephrology
Journal Club presentation of the peer reviewed paper: Alteration of Marrow Cell Gene Expression, Protein Production and Engraftment into Lung by Lung-derived Microvesicles: A Novel Mechanism for Phenotype Modulation
Aliotta JM, Sanchez-Guijo FM, Dooner GJ, Johnson KW, Dooner MS, Greer KA, Greer D, Pimentel J, Kolankiewicz LM, Puente N, Faradyan S, Ferland P, Bearer EL, Passero MA, Adedi M, Colvin GA, Quesenberry PJ.
Stem Cells. 2007 Jul 2
Sanja Selak of Intercell AG, Vienna, Austria, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Intercell develops vaccines for the prevention and treatment of infectious diseases
Gene Olinger, USAMRIID, Fort Detrick USA, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Protective Immune Reponses to Ebola Virus
Paludisme grave : pourquoi doit-on développer des modèles in vitro sur le terrain ? - Conférence de la 8e édition du Cours international « Atelier Paludisme » - RAZAKANDRAINIBE Romy - Madagascar - romy@pasteur.mg
ProImmune Antigen Characterization Summit Paul Mossamandacturner
Paul Moss, School of Cancer Sciences, Birmingham UK, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Cytomegalovirus and Cancer-specific Immunity
Non-Viral Φc31 Integrase Mediated In Vivo Gene Delivery to the Adult Murine Kidney by Daniel C Chung, Matthew C Canver, Xiaofeng Zuo and Jean Bennett and Joshua H Lipschutz* in Experimental Techniques in Urology & Nephrology
Journal Club presentation of the peer reviewed paper: Alteration of Marrow Cell Gene Expression, Protein Production and Engraftment into Lung by Lung-derived Microvesicles: A Novel Mechanism for Phenotype Modulation
Aliotta JM, Sanchez-Guijo FM, Dooner GJ, Johnson KW, Dooner MS, Greer KA, Greer D, Pimentel J, Kolankiewicz LM, Puente N, Faradyan S, Ferland P, Bearer EL, Passero MA, Adedi M, Colvin GA, Quesenberry PJ.
Stem Cells. 2007 Jul 2
After the intravenous transplantation of MSCs, a significant population of cells accumulates in the lung, which they alongside immunomodulatory effect could protect alveolar epithelial cells, reclaim the pulmonary microenvironment, prevent pulmonary fibrosis, and cure lung dysfunction. The fact that the transplantation of MSCs improved the outcome of COVID-2019 patients may be due to regulating inflammatory response and promoting tissue repair and regeneration. This is a preliminary report of our study in Iran.
The NICB (National Institute for Cellular Biotechnology) is located on the Dublin City University (DCU) campus in Dublin, Ireland. It is a leading multidisciplinary centre of translational research in fundamental and applied cellular biotechnology, molecular cell Biology, ocular diseases and biological chemistry. It includes a multidisciplinary team of Cell and Molecular Biologists, Biotechnologists, Chemists and Informatics specialists.
The NICB prioritises translational research involving collaborations with industry and with clinicians, and is committed to educating people from all backgrounds in the area of Biomedical Science.
This slideshare summarises the main research areas of the NICB, including:
Molecular basis for biopharmaceutical production by animal cells
Cancer – drug resistance, invasion and biomarkers
Tissue Engineering/Stem Cell Therapy – ocular diseases, diabetes
Using animal cells as research tools and models for disease research
A presentation on the Stem Cells 21 - IntelliHealthPlus medical center in Bangkok, Thailand. Information on Umbilical cord mesenchymal stem cells and Cd34+ cells, also the companies Ultrasound adipose stem cell separation.
Los días 20 y 21 de octubre de 2016, la Fundacion Ramón Areces organizó un simposio internacional para analizar las 'Enfermedades raras de la piel: de la clínica al gen y viceversa'. El doctor Fernando Larcher Laguzzi, del CIEMAT-Universidad Carlos III de Madrid-IIS Fundación Jiménez Díaz, ejerció de coordinador.
Stem Cells in A New Era of Cell based Therapies - Creative BiolabsCreative-Biolabs
A stem cell can replicate itself or differentiate into cells that carry out the specific functions of the body. The application of stem cells in regenerative medicine and disease therapeutics is one of the most exciting advances in medical science today. In cell-based therapies, stem cells may play two roles. The first role is as drug-delivery vehicles. The second role is as therapeutic agents themselves. Stem cells also offer opportunities for scientific advances that go far beyond cell-based therapies. Creative Biolabs is dedicated to facilitate the research of stem cells in both basic science and therapeutics development. Please contact us if you are interested in our services or products.
Mesenchymal stem cells and their use in inflammatory bowel illnessIJEACS
New pharmacological, surgical, and endoscopic therapies have recently been developed to treat IBD. Among them, stem cell treatment is still in its early stages, despite the fact that multiple studies show that stem cell therapy's immunomodulatory function may decrease inflammation and tissue harm in IBD patients. Intralesional transplantation of autologous or allogeneic MSCs can be deemed a safe and successful therapeutic method for mending perianal fistulas in CD patients, according to this research, which analyses randomized clinical trials and their potential relevance. We did a thorough search of the literature to find research that looked into the function of stem cell treatment in IBD. Since multiple clinical trials have documented exacerbations of IBD following intravenous infusion of MSCs, this literature raises safety concerns about the systemic administration of MSCs.
Paratuberculosis (PTB) remains one of the most obstacles limit animal breeding sector all over the world. The current study aimed to detect the etiology of PTB in tissues of clinically suspected small ruminants using histopathological and real-time polymerase chain reaction (RT-PCR) methods. Clinical examination showed 10 (26.4%) PTB suspected cases out of the total (38) examined animals. The suspected cases were euthanized, necropsied, gross lesions were recorded and tissue samples were collected for histopathological and molecular procedures. Grossly intestinal and mesenteric lymph nodes thickening, corrugations and edematous swellings were recorded. Semi-thin sections of the intestine and mesenteric lymph nodes stained with toluidine blue demonstrated MAP organism inside epithelium cells and macrophages. RT-PCR detected MAP IS900 gene in all suspected cases (100%), thus we recommend using RT-PCR as a rapid sensitive method in the diagnosis of PTB.
Key-words: Paratuberculosis, Mycobacterium, Semi thin sections, Toluidine blue, IS900 gene
Key Trends Shaping the Future of Infrastructure.pdfCheryl Hung
Keynote at DIGIT West Expo, Glasgow on 29 May 2024.
Cheryl Hung, ochery.com
Sr Director, Infrastructure Ecosystem, Arm.
The key trends across hardware, cloud and open-source; exploring how these areas are likely to mature and develop over the short and long-term, and then considering how organisations can position themselves to adapt and thrive.
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
Clients don’t know what they don’t know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clients’ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Securing your Kubernetes cluster_ a step-by-step guide to success !KatiaHIMEUR1
Today, after several years of existence, an extremely active community and an ultra-dynamic ecosystem, Kubernetes has established itself as the de facto standard in container orchestration. Thanks to a wide range of managed services, it has never been so easy to set up a ready-to-use Kubernetes cluster.
However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
Dev Dives: Train smarter, not harder – active learning and UiPath LLMs for do...UiPathCommunity
💥 Speed, accuracy, and scaling – discover the superpowers of GenAI in action with UiPath Document Understanding and Communications Mining™:
See how to accelerate model training and optimize model performance with active learning
Learn about the latest enhancements to out-of-the-box document processing – with little to no training required
Get an exclusive demo of the new family of UiPath LLMs – GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
Speakers:
👨🏫 Andras Palfi, Senior Product Manager, UiPath
👩🏫 Lenka Dulovicova, Product Program Manager, UiPath
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
Slack (or Teams) Automation for Bonterra Impact Management (fka Social Soluti...Jeffrey Haguewood
Sidekick Solutions uses Bonterra Impact Management (fka Social Solutions Apricot) and automation solutions to integrate data for business workflows.
We believe integration and automation are essential to user experience and the promise of efficient work through technology. Automation is the critical ingredient to realizing that full vision. We develop integration products and services for Bonterra Case Management software to support the deployment of automations for a variety of use cases.
This video focuses on the notifications, alerts, and approval requests using Slack for Bonterra Impact Management. The solutions covered in this webinar can also be deployed for Microsoft Teams.
Interested in deploying notification automations for Bonterra Impact Management? Contact us at sales@sidekicksolutionsllc.com to discuss next steps.
Epistemic Interaction - tuning interfaces to provide information for AI supportAlan Dix
Paper presented at SYNERGY workshop at AVI 2024, Genoa, Italy. 3rd June 2024
https://alandix.com/academic/papers/synergy2024-epistemic/
As machine learning integrates deeper into human-computer interactions, the concept of epistemic interaction emerges, aiming to refine these interactions to enhance system adaptability. This approach encourages minor, intentional adjustments in user behaviour to enrich the data available for system learning. This paper introduces epistemic interaction within the context of human-system communication, illustrating how deliberate interaction design can improve system understanding and adaptation. Through concrete examples, we demonstrate the potential of epistemic interaction to significantly advance human-computer interaction by leveraging intuitive human communication strategies to inform system design and functionality, offering a novel pathway for enriching user-system engagements.
Generating a custom Ruby SDK for your web service or Rails API using Smithyg2nightmarescribd
Have you ever wanted a Ruby client API to communicate with your web service? Smithy is a protocol-agnostic language for defining services and SDKs. Smithy Ruby is an implementation of Smithy that generates a Ruby SDK using a Smithy model. In this talk, we will explore Smithy and Smithy Ruby to learn how to generate custom feature-rich SDKs that can communicate with any web service, such as a Rails JSON API.
2. 2
Experimental
Laboratory
GMP Facility
• Cell Production Laboratory
• QC Laboratoy
Preclinical Laboratory
(large animals)
q Regenerative Medicine
q Immunobiology
q Molecular Medicine
q Biomedical Technologies
Advanced therapies
Department of Laboratory Medicine and Advanced Biotechnologies
Laboratory of Clinical Pathology,
Microbiology and Virology
Adoptive Immunotherapies
Transplantation of fetal hepatocytes
Transplantation of pancreatic islets
3. 3
ü Immunotherapeutic approaches (CTLs, NK, dendritic
cells) for treating viral infections and virus-related
tumors
ü Development of regenerative therapies for treating
chronic liver diseases
ü Characterization of stem/progenitors cells in fetal
and adult tissues/organs
ü Characterization of soluble factors produced by stem
cells
Competence
4. GMP Facility - ISMETT
4
Unit of Regenerative Medicine and Biomedical Technologies
5. Unit of Regenerative Medicine and Biomedical Technologies
Organization and Personnel
5
Laboratory of experimental research
n.12 Researchers, n.2 Technicians
(supported by RiMED Foundation)
GMP Facility
(QP, QA, QC, Validation Manager, Production Head)
ISMETT Labs
Laboratory of molecular biology
Laboratory of microbiology
Laboratory of immunology
Cytofluorimetry – Mass Spectrometry
Pre-Clinical Research Lab
Animal Facility
n. 1 Veterinarian
n. 1 Bioengineer
6. Technical setting
6
Unit of Regenerative Medicine and Biomedical Technologies
§ Facility for experimental cell biology (primary cell cultures, cell immortalization, phenotypic &
genotypic analysis , 3D cultures, etc.)
§ Facility for cell production (infected/non-infected cells)
§ Facility for molecular biology (NA amplification, gene expression, sequencing, gene vector
production)
§ Facility for experimental and clinical immunology (lymphoid cells purification, activation and
characterization; immunopheno yping; cell sorting)
§ Facility for microbiology/virology (virus production and analysis; bacterial/yeast cultures for
genetic engineering; control of microbial contamination)
§ Mass spectrometry (proteomics, metabolomics)
§ Biobanking (lymphoid cells for therapy, stem/progenitor cells, primary cell cultures, clinical samples
etc.)
§ Clinical setting for phase I/III studies (cohorts of patients affected by end-stage diseases and
solid organ transplants, highly-specializedsurgical and interventional procedures)
7. Services currently offered
Ø Development of adoptive immunotherapies:
Ø Ag-specific T Cell therapy – Innate immunity–mediated therapy
Ø Production of stem cells in GMP Facility
Ø Mass spectrometry (qualitative/quantitative) analysis of stem cells
secreted factors
Ø Biobanking of primary cultures of adult/fetal human tissues and
pathologic samples of patients affected by end-stage organ
diseases and serious viral/microbial infections
Ø QA assistance for GMP- compliant Standard Operative Procedures
Ø Clinical trials (phase I/II studies)
7
8. Ex vivo production and in vivo infusion of
autologous (or heterologous) cytotoxic/helper
T lymphocyte clones specific to
EBV
CMV
Adoptive cell immunotherapy for prevention and treatment of
post-transplantation herpesvirus infections
Prevention and
treatment of PTLD
Treatment of CMV infections
caused by drug-resistant viral
variants
Trapianti – Infettivologia - Immunoterapia
8
9. ITA 005 colon - treatment with anti-EBV-CTLs
before treatment after treatment
CD20+ B cellsCD20+ B cells
EBEREBER
In vitro production of EBV-specific T cells
Immunoterapia adottiva
CD20+ B cells
EBER
60 Gy
irradiation
9
10. Immunoterapia adottiva per il trattamento di patologie
indotte da herpesvirus in pazienti immunocompromessi
§ Autorizzazione dell’AIFA per il trattamento delle patologie EBV e CMV-correlate nei
pazienti trapiantati
QP/QA: QC: Controllo impianti:
§ Uso di CTL eterologhe anti-EBV e anti-CMV
§ Biomarkers diagnostici di trasformazione EBV- indotta su linfociti B
§ Linee guida di diagnostica molecolare, immunologica e immunoistochimica per
trattamenti terapeutici virus-specifici in fase sintomatica e pre-emptive
(Gruppo di Lavoro su Infezioni in Trapianto: AMCLI-SIV)
§ Partecipazione di ISMETT (socio fondatore) a EATRIS
10
Workshop: L’Infrastruttura di Ricerca EATRIS e il Nodo Italiano:
Stato di avanzamento e prospettive
26-27 Febbraio 2013 - Istituto Superiore di Sanità
The EATRIS consortium of academic institutes provides cutting edge
infrastructure and expertise along the entire translational value chain.
With top quality facilities for basic research, manufacture, non-clinical and
clinical development, our EATRIS institutes offer a truly multidisciplinary
environment.
Prfoducts:
Advanced Therapy Medicinal Products – Biomarkers - Imaging and
Tracing - Small Molecules - Vaccines
11. Isolation of Buffy coat
from liver perfusate
Leukapheresis
Hepatic Lymph nodes
PBMC
from HCV+ patients and
healthy donors
B cell
B cell
HCV-specific
Neutralizing Antibodies
B cell
B cellB cell
EBV- Immortalized
HCV-specific B cells
IV infusion
activated NK
IL2/IFNα/IL12
Low viremic Recipient
(anhepatic phase)
Adapted from: Ohira M et al JCI 2009,119:11 3226-3235
Depletion of CD3+ cells
Isolation of CD56+ NK cells
CliniMACS
Adoptive immunotherapy plan against HCV recurrence
after liver tranplantation
12. Liver perfusate (whole hepatic blood) processing
Infusion in Liver
Recipient
Transfer in 500ml conical
Centrifugation
In vitro NK cells
Activation/expansion
3 X ~3Lt
Reservoirs
Volume reduction
7-10 Lt → 0.5-1.5 Lt
buffy coat
Isolation
Apheresis COM.Tec
Transfer in double
blood bag
Purification of
CD3-CD56+ NK cells
CliniMACS
Clinical grade
MagMACS
research grade
GMP-conforming
12
13. 13
Isolamento Isole di Langerhans
Programma di trapianto insule pancreatiche
- trapianto eterologo, autotrapianto –
14. Insule pancreatiche – diabete
14
Nano Engineering for Cross Tolerance: New approaches for bioengineered, vascularized,
chimeric islet transplantation in non-immunosuppressed hosts (NEXT)
Progetto presentato per FP7 HEALTH.2013.1.3-2: Innovative approaches to address
adverse immune reactions to biomedical devices, implants and transplant tissues -
Coordinator dr. Severino – Explora srl
Studio di piccole molecole cito-protettive con duplice applicabilità nella demenza di Alzheimer
e nel trattamento del diabete mediante trapianto di insule pancreatiche
Progetto finanziato PON 02- 00697 Potenziamento laboratori pubblico-privati
In collaborazione con CNR IBB di Catania, RiMED, Myrmex SpA (ccordinatore CNR –IBB)l
15. Transplantion of human fetal liver cells in patients
with end-stage liver cirrhosis
Fetal liver cell
suspension
Collagenase digestion
Infusion into patient
splenic arteryHuman fetal
liver
Phase I-II Control Study of Human Fetal Liver Cell Transplantation for
Treatment of Chronic Liver Disease
(Submitted manuscript)
Efficient human fetal liver cell isolation protocol based on
vascular perfusion for liver cell-based therapy and case report on
cell transplantation
(Liver Transpl. 18:226-237, 2012)
15
16. Human fetal hepatocytes cultured in high density
demonstrate mature hepatic functionality
*
0
50
100
150
200
250Conc(ng/ml)/1.8x10
6
cells
Albumin secretion
Fetal Adult Liver
MSCs
0
2
4
6
8
10
12
14
16
18
Conc(mg/dL)/1.8x10
6
cells
Urea production
Fetal Adult Liver
MSCs
0
50000
100000
150000
200000
250000
300000
RLU/1.8x106
cells
CYP3A4 activity
Fetal Adult
Liver
MSCs
G6Pase activity Glycogen storage ICG uptake
Primary culture of
fetal hepatocytes
High density culture Low density culture Flat cell culture
1:3 split
0
50
100
150
200
250
Conc(ng/ml)/1.5x10
6
cells
Down-regulation albumin
secretion in flat cell cultures
Fetal hep. Flat
cells
Liver
MSCs
*
Cryopreservation up to 1 year storage is well tolerated as did not significantly affect viability (A), and
functions (B, C) of fetal hepatocytes
60
65
70
75
80
85
90
FRESH CRYO
Percentageofviability
Fresh Cryo
10
30
50
70
90
110
130
150
170
190
F R ES H C R Y O
Conc.(ng/ml)/1.8x10
6
cells
Fresh
Cryo
A B
C
Cryo
16
17. 17
Medicina rigenerativa: cellule staminali dermiche
Ø Multipotent fetal dermal cells are easy to isolate, with a high yield
Ø Cultured fetal dermal stem cells possess a mesenchymal phenotype
Ø Cells with a regenerative potential can be successful isolated from a
small fetal skin biopsy and maintained in culture for long periods without
losing their potential, thus generating large quantities for clinical
applications.
18. 18
In vitro Expansion and Characterization of Skin
Precursor Cells Isolated from Human Fetal Dermis
CD71
CD105
dermal
papilla dermal
papilla
A B C
CD105
CD90 CD73 CD105
Fetal Adult
SSEA4SSEA4
90% 10.5%
Pluripotency marker SSEA-4 in fetal
and adult dermal cells
Approx. 90% positive for the classical
mesenchymal markers CD90, CD73 and CD105
Localized in close proximity of skin
dermal papillae
Low immunogenicity. (A) Fetal dermal
cells co-cultured with allogenic PBMCs
do not evoke T cell proliferation. (B)
Positive control MLR
A B
A
0
5
10
15
20
25
30
P1
P3
P5
P7
P9
P11
P13
P15
P17
P19
P21
P23
P25
P27
P29
Passage number
Numberofdaystothenext
passage
Adult Fetal
Day 0
CB
Day 6
Retention of a stable phenotype
after 12 subcultivations
Expansion potential of fetal vs. adult dermal cells
(A) In vitro angiogenic potential. (B, C) Epithelial-like
potential DMSO-induced
A B
AD P10Fet P25
Higher retention of differentiation
potential in fetal than adult cells
0
20
40
60
80
100
120
CD90
CD105
CD73
HLAclassI
CD44
CD166
CD71
Vimentin
CD29
Nestin
CD49b
CD49e
CD106
CD117
CD45
CD34
CD14
HLADR
%positivecells
P3
P12
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Cellnumber(106
)
(C) Non enzymatic isolation technique of “cell outgrowth”. (D)
cell yield after isolation: fetal cells can be isolated with a 3-fold
higher efficiency than adult cells
Adult
Fetal
19. Tissue Stem/Progenitor Cells & Microenvironment Modulation
TISSUE OF ORIGIN:
≠ types of tissues ; ≠ age (e.g. fetal and adult tissues)
DEVELOPMENT & OPTIMIZATION of CULTURE SYSTEMS:
2D and 3D ; use of scaffolds ; growth factors + ; hypoxia
Mesenchymal Stem Cells
“Stemnessdegree”
Tuning&Evaluation
Models of assessment of therapeutic and regenerative potentials in vivo
SILAC
Microvescicles
cell-cell contacts
Secreted Factors: Qualitative & Quantitative ProteomicsSecret-OMICS
miRNA
Light AA Heavy AA
Conditioned Media Processing
Quantitative identification of
full differential secretome
LC-MS/MS
Systems Biology
Bioinformatics
Data Functional Analysis
Integration of
Transcriptomics Data
Secreted Factors Mix &
Systems Biology
In Gel
Digestion
Protein identification
Protein quantification