Hepatocyte growth factor and fibroblast growth factor-4-induced differentiation of human bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro.
- Author:
Jin-min XIE
1
;
Jian-feng CHEN
;
Yi GAO
;
Kun-hou YAO
Author Information
- Publication Type:Journal Article
- MeSH: Bone Marrow Cells; cytology; drug effects; metabolism; Cell Differentiation; drug effects; Cells, Cultured; Fibroblast Growth Factor 4; pharmacology; Hepatocyte Growth Factor; pharmacology; Hepatocytes; cytology; drug effects; metabolism; Humans; Immunohistochemistry; Keratin-18; biosynthesis; Mesenchymal Stromal Cells; cytology; drug effects; metabolism; RNA, Messenger; genetics; metabolism; Reverse Transcriptase Polymerase Chain Reaction; alpha-Fetoproteins; biosynthesis; genetics
- From: Journal of Southern Medical University 2006;26(10):1439-1442
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo induce the differentiation of human bone marrow mesenchymal stem cells (HMSCs) into hepatocyte-like cells with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro.
METHODSHMSCs were induced to differentiate into hepatocyte-like cells by HGF (group B), FGF-4 (group C) and HGF+FGF-4 (group D) in vitro. Undifferentiated HMSCs and L-02 cells were used as the negative (group A) and positive (group E) controls, respectively. The changes of cell morphology were observed microscopically. The expressions of hepatic markers, alpha fetoprotein (AFP) and CK-18, were detected by immunocytochemical staining at different times after induction, and the differentiation ratios of the various groups of HMSCs were calculated on the basis of image analysis. The expressions of AFP and ALB were detected by immunofluorescence assay in each group at different times after induction, and the expressions of AFP and ALB mRNA by RT-PCR.
RESULTSHMSCs gradually transformed into spindle-shaped, round, polygonal or irregular cells after induction. Immunocytochemical staining revealed positive AFP and CK18 expressions in groups B, C, and D after induction as well as in group E. The positive units (PU) of AFP and CK18 in group D calculated according to image analysis were significantly higher than that of groups A, B, and C. The expressions of AFP and ALB detected by immunofluorescence were both positive after induction in all groups except group A, similar to the findings of the expressions of AFP and ALB mRNA by RT-PCR.
CONCLUSIONHMSCs can be induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and their combination at certain concentrations, and the hepatocyte-like cells can express some hepatic markers such as AFP, ALB, CK18, etc. HGF+FGF-4 may achieve more effective induction of HMSC differentiation into hepatocyte-like cells, and the efficiency of HGF is greater than that of FGF-4.
