The Bionomics, Plasticity And Hepatogenic Differentiation Capacity Of The Adipose Derived Stem Cells Form Donors With Chronic Liver Diseases And Health Adults

The development of tissue engineering has been restricted by the source of seed cells.Since adipose derived stem cells(ADSCs)were discovered,more and more studies confirmed that these cells could be induced to adipose cells,cartilage cells,myocyte and osteblasts,etc,by specific cell factors.The ADSCs not only have the ability of multi-directional differentiation,but also can excrete the transforming growth factor-β(TGF—β)and hepatocyte growth factor(HGF), both of which can inhibit the cell immunity of the host.So the ADSCs can be used to treat some inflammatory diseases as well as release the rejection of host in the allograft transplantation.Adipose tissue is easy to be obtained and autotransplant. Furthermore,ADSCs could keep the potential of multi-directional differentiation when they are cultured in vitro.In addition the genetic background ADSCs are very stable.Because of these advantages,they are suitable seed cells for tissue engineering.However the biological behavior of ADSCs such as metabolic capability,proliferation and differentiation potency could be greatly influenced by the origin position of the adipose tissue,age and sex of the donors.The biology and plasticity of ADSCs from different donors with or without chronic liver disease were investigated to provide theroretical basis for further application of the ADSCsPART 1Objective:To study the morphology,growth,adhesion,proliferation,cell surface marker of ADSCs of different donors.Method:20 donors were allocated to 2 groups:10 donors without chronic diseases and 10 donors with the history of type B hepatitis for more than 10 years and the history of liver cirrhosis.Then each group was further divided into 2 groups according to their sex.The 4 groups were:Group A:male,41-60years old,without chronic disease;Group B:female, 43-58 years old,without chronic disease;Group C:male,45-63 years old,with the chronic hepatic disease;Group D female,41-61 years old,with chronic hepatic disease.The ADSCs were isolated from the abdominal wall cell subcutaneous adipose tissue of the donors;the cell morphology was observed by using inverted microscope;cell surface markers were examined by immunohistochemistry and then the percent of positive cells was analyzed in image-pro plus4.0;growth character was determined by cell counting.Reault:ADSCs from different donors showed the same long fusiform shape.About 80%cells are positive for CD13, CD44 and CD105;Time-growth curve and doubling time of ADSCs differed in groups with or without chronic liver disease.Conclusion:ADSCs had many advatages,such as easily to be isolated and cultured,stationary proliferation rate, etc.ADSCs from donors with chronic liver disease had lower proliferation rate than ADSCs from ones without chronic disease.PART 2Objective:To study the plasticity of ADSCs from different donors.Method: Inducing multi-differentiation of one donor with chronic hepatic disease and one healthy.①ADSCs were induced to osteoblasts by water-solubility dexamethasone andβ—glycerophosphate.AKP stain was made after 21 days.②They were induced to adipose cells by isobutyl 1- methylxanthine(IBMX),water-solubility dexamethasone,insulin and indomethacin.Oil Red stain was made after 14 days induction.③They were induced to cardiomyocyte-like cells by 5-azacytidine,α—Actin stain was made after 24h induction.Results:The induced cells were positive for AKP after 21d induction;positive in Oil Red O stain after 14d induction;positive inα—Actin stain after 24h induction.Conclusion:ADSCs from different donors could be induced to ostoblasts,adipose cells, cardiomyocyte-like cells,and the plasticity was good.PART 3Objective:To study the ability of being induced to the hepatocyte-like cells of the ADSCs from different donors.Method:ADSCs were induced to hepatocyte-like cells by HGF,OSM and DMSO.The expression of AFP and ALB were examined by RT-PCR and flow cytometry after 14d induction.Results: When the age and sex were matched,the ADSCs from group C and group D showed lower induction rate and ALB gene expression than the ADSCs from group A and group B.Conclusion:The ADSCs from donors with chronic liver disease showed lower ability of being induced into hepatocyte-like cells than those from healthy donors.