This study investigated the proliferation characteristics and tumorigenic potential of oval cells, the proposed stem cells of the liver. Male rats were fed a choline-deficient diet to induce oval cells, which were then isolated and cultured over 100 passages over 2 years. The oval cells were analyzed using flow cytometry and markers to observe their immunophenotype, proliferation ability, and differentiation potential. When xenografted into immunodeficient mice, the oval cells did not spontaneously transform into cancer-initiating cells. This suggests hepatic oval cells may serve as a safe cell source for future cell therapy and studying liver carcinogenesis.
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clini...IOSR Journals
Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from csection with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively. Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine, Deoxycholate, PPARG, C/EBPα, etc.
Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure.
Meritxell Huch - Wellcome Trust/Cancer Research UK Gurdon Institute, Universi...Fundación Ramón Areces
El jueves 8 de febrero de 2018 se realizó en la Fundación Ramón Areces un Ciclo de Conferencias sobre células madre y organoides, en colaboración con Springer Nature.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
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.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clini...IOSR Journals
Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from csection with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively. Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine, Deoxycholate, PPARG, C/EBPα, etc.
Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure.
Meritxell Huch - Wellcome Trust/Cancer Research UK Gurdon Institute, Universi...Fundación Ramón Areces
El jueves 8 de febrero de 2018 se realizó en la Fundación Ramón Areces un Ciclo de Conferencias sobre células madre y organoides, en colaboración con Springer Nature.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
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.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Induced Pluripotent Stem-Like Cells Derived from Ban, a Vietnamese Native Pig...AI Publications
Induced pluripotent stem cells (iPSc) is a promising technology for applying in bio-medicine and biodiversity conservation. In the present study, we isolate and culture fibroblasts from Ban – a Vietnamese native pig breed and transfer episomal plasmid containing genes Oct3/4, Sox2, Klf4, l-Myc, LIN28 and EBNA1 in order to reprogram cells. We isolated, cultured and cryopreserved successfully 9 primary fibroblast lines from Ban (culture percentage is 90.0%). Plasmids was successfully transferred into Ban fibroblasts with high efficiency. Changes in morphology of fibroblasts into pluripotent stem-like cells showed that they had been reprogrammed under the effect of transferred genes. The pluripotency signal was further proved by in vitro differentiation by formation of embryoid body in all 3 transfected cell lines. The results showed that pluripotent stem-like cells has successfully derived in Ban pigs.
Poster – Development and automation of 3D innovative hiPSC-based liver organo...HCS Pharma
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
Poster - A single procedure to generate functional hiPSCs-derived liver organ...HCS Pharma
metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
Induced Pluripotent Stem-Like Cells Derived from Ban, a Vietnamese Native Pig...AI Publications
Induced pluripotent stem cells (iPSc) is a promising technology for applying in bio-medicine and biodiversity conservation. In the present study, we isolate and culture fibroblasts from Ban – a Vietnamese native pig breed and transfer episomal plasmid containing genes Oct3/4, Sox2, Klf4, l-Myc, LIN28 and EBNA1 in order to reprogram cells. We isolated, cultured and cryopreserved successfully 9 primary fibroblast lines from Ban (culture percentage is 90.0%). Plasmids was successfully transferred into Ban fibroblasts with high efficiency. Changes in morphology of fibroblasts into pluripotent stem-like cells showed that they had been reprogrammed under the effect of transferred genes. The pluripotency signal was further proved by in vitro differentiation by formation of embryoid body in all 3 transfected cell lines. The results showed that pluripotent stem-like cells has successfully derived in Ban pigs.
Poster – Development and automation of 3D innovative hiPSC-based liver organo...HCS Pharma
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
Poster - A single procedure to generate functional hiPSCs-derived liver organ...HCS Pharma
metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
Higher expression of SATB2 in hepatocellular carcinoma of African Americans d...rakeshsrivastava89
Abstract
In the United States, Hepatocellular Carcinoma (HCC) incidence has tripled over the past two decades. The disease has disproportionately affected minority and disadvantaged
populations. The purpose of this study was to examine the expression of SATB2 gene in HCC cells derived from African Americans (AA) and Caucasian Americans (CA) and assess its oncogenic potential by measuring cell viability, spheroid
formation, epithelial‐mesenchymal transition (EMT), stem cell markers and pluripotency maintaining factors in cancer stem cells (CSCs). We compared the expression of SATB2 in human primary hepatocytes, HCC cells derived from AA and CA, and
HCC CSCs. Hepatocellular carcinoma cells derived from AA expressed the higher level of SATB2 than those from CA. By comparison, normal human hepatocytes did
not express SATB2. Higher expression of SATB2 in HCC cells from AA was associated with greater growth rate, cell viability, colony formation and EMT characteristics than those from CA. Knockout of SATB2 in CSCs by Crispr/Cas9 technique significantly inhibited the expression of SATB2 gene, stem cell markers (CD24, CD44 and CD133), pluripotency maintaining factors (c‐Myc, KLF4, SOX2 and OCT4), and EMT
compared with non‐targeting control group. The expression of SATB2 was negatively correlated with miR34a. SATB2 rescued the miR‐34a‐mediated inhibition of CSC's viability. These data suggest that SATB2 is an oncogenic factor, and its higher expression may explain the disparity in HCC outcomes among AA.
Wei Yu1 | Sanjit K. Roy2 | Yiming Ma1 | Thomas A. LaVeist3 | Sharmila Shankar1,2,4 |
Rakesh K. Srivastava1,2,4
Brucea javanica oil inhibits proliferation of hepatocellular carcinoma cells ...LucyPi1
Abstract Background: Brucea javanica oil (BJO), distributed primarily in Southeast Asia, has long been utilized as a therapeutic agent for treating malignancies. However, its anticancer mechanisms are not clearly understood. The objective of this study was to examine the mechanisms underlying its treatment of hepatocellular carcinoma cells. Methods: CCK8 assay was used to evaluate cell viability. Hoechst33342 staining and flow cytometry analyses were used to examine apoptosis. Mito-Tracker Red CMXRos kit was used to measure the membrane potential of mitochondria. ATP assay kit was used to evaluate ATP levels. Western blots were used to assess the presence of AKT, adenosine monophosphate-activated protein kinase, Caspase3, Caspase9, Bax, and Bcl-2. Results: BJO inhibited the proliferation of hepatocellular carcinoma cells HepG2 in a time- and dose-dependent manner. It induced apoptosis, with the percentage of cells treated with 50–150 μg/mL BJO increasing from 8.01% to 28.02% in a concentration-dependent manner (P < 0.05, when 50 μg/mL of BJO group compared with the control group; P < 0.001, when 100 or 150 μg/mL of BJO group compared with the control group). After exposed to BJO, the expression of C-caspase3, C-caspase9 and Bax upregulated while that of Bcl-2 downregulated. BJO suppressed the PI3K/AKT pathway and promoted phosphorylation of adenosine monophosphate-activated protein kinase, while repressing the phosphorylation of mechanistic target of rapamycin. Compared with treatment by BJO alone, the PI3K/AKT agonist 740Y-P increased the survival rate of HepG2 cells (P < 0.01) and attenuated the inhibitory effect of BJO on cell apoptosis (P < 0.05). Conclusion: BJO is capable of inhibiting proliferation of HepG2 cells and inducing apoptosis via the PI3K/AKT pathway.
Evaluation of hepatoprotective agents - Hemant KanaseHemant Kanase
1. Introduction
2. Hepatotoxicity: Mechanism
3. Therapeutic strategies available – their limitations
4. In vivo models of liver damage
- Non-invasive model
a. Chemically induced hepatotoxicity
b. Drug-induced hepatotoxicity
c. Radiation-induced hepatotoxicity
d. Metal-induced hepatotoxicity
e. Diet-induced hepatotoxicity
Models of Acute Hepatitis
Models of chronic hepatitis
Models of fibrosis
Models of cholestasis
Models of steatosis
4. Problems faced with animal studies
5. In vitro models of liver damage
6. Advantages and disadvantages of in vitro models
7. Parameters of evaluation
8. Clinical Assessment
Kaempferol increases levels of coenzyme Q in kidney cells and serves as a biosynthetic ring precursor
Complete study available in Free Radical Biology and Medicine. 2017 Sep;110:176-187.
doi: 10.1016/j.freeradbiomed.2017.06.006. Epub 2017 Jun 9.
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
Toxic effects of heavy metals : Lead and Arsenicsanjana502982
Heavy metals are naturally occuring metallic chemical elements that have relatively high density, and are toxic at even low concentrations. All toxic metals are termed as heavy metals irrespective of their atomic mass and density, eg. arsenic, lead, mercury, cadmium, thallium, chromium, etc.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptxMAGOTI ERNEST
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
DERIVATION OF MODIFIED BERNOULLI EQUATION WITH VISCOUS EFFECTS AND TERMINAL V...Wasswaderrick3
In this book, we use conservation of energy techniques on a fluid element to derive the Modified Bernoulli equation of flow with viscous or friction effects. We derive the general equation of flow/ velocity and then from this we derive the Pouiselle flow equation, the transition flow equation and the turbulent flow equation. In the situations where there are no viscous effects , the equation reduces to the Bernoulli equation. From experimental results, we are able to include other terms in the Bernoulli equation. We also look at cases where pressure gradients exist. We use the Modified Bernoulli equation to derive equations of flow rate for pipes of different cross sectional areas connected together. We also extend our techniques of energy conservation to a sphere falling in a viscous medium under the effect of gravity. We demonstrate Stokes equation of terminal velocity and turbulent flow equation. We look at a way of calculating the time taken for a body to fall in a viscous medium. We also look at the general equation of terminal velocity.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
ANAMOLOUS SECONDARY GROWTH IN DICOT ROOTS.pptxRASHMI M G
Abnormal or anomalous secondary growth in plants. It defines secondary growth as an increase in plant girth due to vascular cambium or cork cambium. Anomalous secondary growth does not follow the normal pattern of a single vascular cambium producing xylem internally and phloem externally.
2. Viral hepatitis and liver cancer;M Levrero ;Nature review , 2006
Liver cancer as a result of chronic liver injury
Department of Surgery, University of Tsukuba
3.
4. Study overview
Department of Surgery, University of Tsukuba
Snapshot :
Oval cell is considered the stem cell of liver .
This study tries to investigate the proliferation characteristics, and
tumorgenesis of oval cells .
Male Sprague dawley rat was feed with choline deficient diet CDE(to induce
oval cell )>>>hepatic oval cell isolation >>2 years culture :100 passage >>
Flow cytometry –immunoflurence ( Ov6 , BD-2, BD-2, Dlk antibodies
/markers)>>microscopic morphology study>proliferation ability > colony
formation ability study >>>
Xenograft into immunodeficient mouse to observe tumorgenesis .
HepG2 cell line was used as control in this study .
5. Male Sprague dawley rat was feed with choline
deficient diet CDE(to induce oval cell )-6 weeks
hepatic oval cell isolation
2 years culture :100 passages
study
Xenograft into immunodeficient
mouse to observe tumorgenesis .
Department of Surgery, University of Tsukuba
Research flow
12. Department of Surgery, University of Tsukuba
Hepatic oval cell proliferation capability and
chromosomal stability
13. Department of Surgery, University of Tsukuba
Colony formation assessment: anchorage growth assessment
14. Department of Surgery, University of Tsukuba
In vivo :xenograft to immunodeficient mouse
15. Department of Surgery, University of Tsukuba
In summary, serial passages in vitro did not change the
immunophenotype,proliferation capacity, or differentiation
potential of hepatic oval cells and did not cause spontaneous
maltransformation of these cells.
Therefore, hepatic oval cells without
immortalized manipulation not only provide an expandable
cell source for future cell-therapy utilizations but also may
serve as a cell model for understanding the mechanism of
carcinogenesis
This study also suggests that hepatic oval cells did not
experience spontaneous maltransformations into cancer-
initiating cells, suggesting the safety of utilizing hepatic
stem/progenitor cells for exploitation of stem cell
technology.
Summary
Editor's Notes
This diagram shows the pathogenesis of liver cancer : Chronic liver injury such as hepatitis B and C infection and alcohol can result liver hepatitis and cirrhosis and eventually cancer , this is a multistep progressive process lead to genetic alteration resulting dysplastic nodeules (preneoplastic nodules ) which later develop into cancer.
Pathogenesis of human hepatocellular carcinoma (HCC). Chronic hepatitis B and C and associated liver cirrhosis represent major risk factors for HCC development, being implicated in more than 70% of HCC cases worldwide. Additional etiological factors, which often represent co-factors of an underlying HBV- or HCV-related chronic liver disease, include toxins and drugs (e.g., alcohol, aflatoxins, microcystin, anabolic steroids), metabolic liver diseases (e.g., hereditary hemochromatosis, alpha1-antitrypsin deficiency), steatosis, non-alcoholic fatty liver diseases and diabetes. Hepatocarcinogenesis is a multistep process that may last for decades and involves the progressive accumulation of different genetic alterations ultimately leading to malignant transformation. Regardless of the etiological agent, malignant transformation of hepatocytes is believed to occur through a pathway of increased liver cell turnover, induced by chronic liver injury and regeneration, in a context of inflammation and oxidative DNA damage. Dysplastic nodules and macroregenerative nodules are considered as pre-neoplastic lesions. The detailed analysis of HCC development in experimental animals and the comparison of the results with HCC in humans has identified a variety of genomic and molecular alterations in fully developed HCC and to a lesser extent in morphologically defined pre-neoplastic precursor lesions. At least four pathways that regulate either cell proliferation or cell death (i.e., the phospho-retinoblastoma (pRb), p53, transforming growth factor-beta (TGF-beta) and beta-catenin pathways) are affected in HCCs.
Journal of Gastroenterology and Hepatology (2003) , REVIEW Oval cell-mediated liver regeneration: Role of cytokines and growth factors
The 2-AFF induced the proliferation of a periportal population of oval like cells ( Negative for expression of classical oval cell markers ) .However , the expression of these markers (AFP) started after two days ( Hepato-lineage ) affliated subpopulation .
Background & Aims: Although expandable hepatic progenitors
provide renewable cell sources for treatment of hepatic disorders,
long-term cultivation of hepatic progenitors may affect proliferation
and differentiation abilities, and even initiate the formation
of malignant cancer stem cells. This study aims to determine
characteristics of primary cultured hepatic oval cells after prolonged
cultivation in vitro.
This will be the steps this experiment.
1. The typical epithelial morphology of hepatic oval cells after serial passages. (A) The morphology of hepatic oval cells under low phase-contrast microscope
Fig. 2. Flow cytometry analysis demonstrating that hepatic oval cells expressed hepatic progenitor cell markers. The positive rate of the common marker for hepatic
oval cells, OV-6, at primary culture, 10th, 50th, and 100th passage, was 87.2%, 77.7%, 81.1%, and 76.7%, respectively. The expression of BD-1, BD-2, and Dlk, was over 85%,
90%, and 98%, respectively. As a negative control, primary cultured hepatocytes did not express these hepatic progenitor cell markers.
Fig. 3. Hepatic oval cells sustained expression of AFP, ALB, and CK19. (A) Immunofluorescence results showed that hepatic oval cells at the 0–2nd, 10th,
50th, and 100th passage expressed hepatocytic markers AFP and ALB. (B) Realtime RT-PCR results showed that hepatic oval cells had a sustained expression of
AFP, ALB, and CK19. (C) Western blot results demonstrated that in hepatic oval cells the expression of AFP, ALB, and CK19 persisted.
Fig. 4. Hepatocyte-specific differentiation capacity of hepatic oval cells after serial passages. (A) High power phase contrast images of hepatic oval cells exposed to
sodium butyrate (0.75 mM) for 3 days, and stained for Giemsa or PAS; hepatic oval cells at different passages increase in cell size, become binuclear and/or polyploid and
accumulate intracellular polysaccharides. Periodic acid–Schiff (PAS) is a staining method used to detect polysaccharides such as glycogen, and mucosubstances such as glycoproteins, glycolipids and mucins in tissues. (B) Western blot showing that the expression of ALB is upregulated, CK19 is slightly downregulated, while CK18 is significantly
decreased after sodium butyrate (0.75 mM) inducement from different passages of oval cells. (C and D) Quantification of the albumin secretion and ureagenesis showing
that albumin in the supernatant is increased and transformed urea is also increased at different passages of oval cells after sodium butyrate (0.75 mM) treatment.
B) Western blot showing that the expression of ALB is upregulated, CK19 is slightly downregulated, while CK18 is significantly
decreased after sodium butyrate (0.75 mM) inducement from different passages of oval cells. (C and D) Quantification of the albumin secretion and ureagenesis showing
that albumin in the supernatant is increased and transformed urea is also increased at different passages of oval cells after sodium butyrate (0.75 mM) treatment.
Fig. 5. Hepatic oval cells survive after serial passages without loss of their proliferation ability and maintain diploid cells with features of chromosomal
stability. (A) Growth curves for the cell count of hepatic oval cells revealed that their doubling time at the 0–2nd, 10th, 50th, and 100th passage was 25.8, 25.4,
22.3, and 23.1 h, respectively. (B) Representative flow cytometry analysis of DNA content and cell cycle showing that hepatic oval cells sustain their proliferation
potential and maintain the typical PI-staining pattern for diploid cells.
Fig. 6. Colony formation in vitro and tumorigenesis in vivo. (A) HepG2 cells and hepatic oval cells at different passages plated in soft agar for 3 weeks. Unlike the positive
control, tumorigenic HepG2 cells, hepatic oval cells at different passages fail to proliferate in soft agar. (B and C) Quantification by ELISPOT of the rate of formation of
colonies (B), and the % of the well area covered (C) (*p <0.05) for hepatic oval cells, there is no significant difference in colony number and covered area at the 0–2nd, 10th,
50th, and 100th passage. (D) In vivo tumorigenesis by injection of different passages of oval cells subcutaneously in the left flank of the immunodeficient mouse (five mice
per group); the positive control cells, HepG2, induce tumors after 2 and 4 weeks in the immunodeficient mice, while the oval cells cause no tumors.
Fig. 6. Colony formation in vitro and tumorigenesis in vivo. (A) HepG2 cells and hepatic oval cells at different passages plated in soft agar for 3 weeks. Unlike the positive
control, tumorigenic HepG2 cells, hepatic oval cells at different passages fail to proliferate in soft agar. (B and C) Quantification by ELISPOT of the rate of formation of
colonies (B), and the % of the well area covered (C) (*p <0.05) for hepatic oval cells, there is no significant difference in colony number and covered area at the 0–2nd, 10th,
50th, and 100th passage. (D) In vivo tumorigenesis by injection of different passages of oval cells subcutaneously in the left flank of the immunodeficient mouse (five mice
per group); the positive control cells, HepG2, induce tumors after 2 and 4 weeks in the immunodeficient mice, while the oval cells cause no tumors.