Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells with the combined induction of hepatocyte growth factor and fibroblast growth factor
10.3969/j.issn.1673-8225.2009.45.001
- VernacularTitle:肝细胞生长因子与成纤维生长因子联合诱导入脐带间充质干细胞向肝样细胞的分化
- Author:
Huamin ZHU
;
Huan ZHANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(45):8811-8816
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Fibroblast growth factor can promote proliferation of mesenchymal stem cells (MSCs).grew along the wall.However,there are few reports on the differentiation of MSCs into hepatocytes following fibroblast growth factor induction.When mass concentration of hepatocyte growth factor was 1 μg/L,it can promote mitosis of hepatocytes,and is the strongest mitogenic agent for normal hepatocytes.OBJECTIVE:To explore the biological characteristics of human umbilical cord MSCs/n vitro and their differentiation ability to hepatocyte-like cells under the induction of chemical factors.DESIGN,TIME AND SETTING:The cytological in vitro study was conducted at the Institute of Blood,Jinan University from August 2008 to April 2009.MATERIALS:Umbilical cord was obtained from healthy full-term fetus,which was provided by the Guangzhou Huaqiao Hospital.The parturient signed informed consent.Hepatocyte growth factor and flbroblast growth factor were bought from Peprotech,USA.METHODS:The MSCs from human cord were isolated and cultured by type Ⅳ colagenase digestion+differential adherence.At the third passage,MSCs received cell surface antigen analysis and cell cycle determination to detect their ability to differentiate into adipocytes and osteoblasts.At the fifth passage,MSCs were adjusted into 5×10~9/L and assigned into 2 groups.BMSCs in the control group were incubated in DMEM/F12 containing 5% fetal bovine serum.BMSCs in the induction group were treated with above-mentioned medium supplemented with 20 μg/L hepatocyte growth factor and 10 μg/L flbroblast growth factor.MAIN OUTCOME MEASURES:The following parameters were measured:biological characteristics of human umbilical cord MSCs and differentiation of human umbilical cord MSCs into hepatocyte-like cells in vitro.RESULTS:At the third passage,MSCs derived from human cord expressed CD29,CD44,CD105,but not antigens of hematopoietic CD34,CD45,and 92.2% of them were in G_0/G_1 phase.Oil red O staining showed cytoplasm presented red granules.Alkaline phosphatase staining demonstrated that cytoplasm was black,with the differentiation ability into adipocytes and osteoblasts.Following 10 days of combined induction of hepstocyte growth factor and fibroblast growth factor,RT-PCR and Western blot results confirmed that cells expressed alpha fetoprotein and albumin.Negative expression was found in the control group.CONCLUSION:Human umbilical cord contained plenty of MSCs,with strong potential of multi-differentiation.Umbilical cord MSCs can differentiate into hepatocyte-like cells following combined induction of hepatocyte growth factor and fibroblast growth factor.
