Differentiation of mesenchymal stem cells from umbilical cord blood into hepatocyte-like cells in the in vitro culture
- VernacularTitle:人脐血间充质干细胞在体外向肝样细胞的诱导分化
- Author:
Zuohua CHI
;
Yuan ZHANG
;
Yan LU
- Publication Type:Journal Article
- Keywords:
Fetal blood;
Mesenchymal stem cells;
Cell differentiation;
Hepatocytes
- From:
Chinese Journal of Organ Transplantation
2008;29(3):159-163
- CountryChina
- Language:Chinese
-
Abstract:
Objective To discuss the possibility of differentiation of mesenchymal stem cells(MSCs)from umbilical cord blood(UCB)into hepatocyte-like cells in the in vitro culture.Methods MSCs were isolated from UCB,cultured and passaged.The surface markers were examined by flow cytometry.When cells were cultured to the third passage,they were inoculated into a 6-well plate.A two-stage induction method was used:MSCs in the first phase were cultured in DMEM/F12 medium supplemented with dexamethasone(final concentration of 0.5 μmol/L,the same below),hepatocyte growth factor(HGF,10 ng/ml),epidermal growth factor(10 ng/m1)and 1×insulin-transferrin-Se(ITS)for two weeks,then in DMEM/F12 supplemented with 0.5 μmol/L dexamethasone,10 ng/ml HGF,1×ITS,10 ng/ml Oncostatin M for another two weeks.Morphological changes were observed under a microscope.The gene expression correlated with hepatocytes was detected by using RT-PCR.Immunofluorescence staining was used to identify the expression of specific protein related to hepatocytes(AFP,Albumin,CK-18).Ultrastructure was detected under an electron microscope.Results In the cultured MSCs from UCB,CD34/CD45/CD14,CD54,CD49f and HLA-DR were not detected,there was low expression of CD106 and strong expression of CD29,CD44 and CD13.The gene expression of AFP,albumin,CK-18 and TAT was discovered and three kinds of protein AFP,albumin and CK-18 were positively showed in cytoplasm after 4 weeks'induction.The hepatin granules and fatty drops in cytoplasm of cells induced for 4 weeks were found under an electron microscope.Conclusion The MSCs fromUCB can differentiate into hepatocyte-like ceils in the in vitro culture under some conditions.
