Differentiation of 5-azacytidine-induced bone marrow stromal stem cells transduced with hepatocyte growth factor gene.
- Author:
Qian WEN
1
;
Chao-ying ZHOU
;
Ming-qian ZHOU
;
Wei LUO
;
Li MA
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Azacitidine; pharmacology; Bone Marrow Cells; cytology; drug effects; metabolism; Cell Differentiation; drug effects; Cells, Cultured; Dogs; Gene Expression; Hepatocyte Growth Factor; genetics; metabolism; Mesenchymal Stromal Cells; cytology; Transfection
- From: Journal of Southern Medical University 2010;30(7):1537-1540
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo examine the differentiation of 5-azacytidine-induced bone marrow stromal stem cells (BMSCs) into cardiomyocyte-like cells and hepatocyte growth factor (HGF) gene expression after HGF gene transfection.
METHODS5-azacytidine was used to induce beagle dog BMSCs to differentiate into cardiomyocyte-like cells. Morphological observation and immunohistochemistry were performed to detect the expression of the markers of cardiomyocyte-like cells including beta-myosin heavy chain (beta-MHC) and alpha-sarcomeric actin. The cells were then transfected with Ad-HGF, and the mRNA and protein expressions of HGF gene were detected by RT-PCR and ELISA, respectively.
RESULTSAfter 4 weeks of induction with 5-azacytidine, the BMSCs differentiated into cardiomyocyte-like cells. The expressions of HGF at the mRNA and protein levels were confirmed in the cells after transfection with Ad-HGF. The peak HGF protein secretion was 10(3) ng/ml at 48 h after transfection.
CONCLUSIONAd-HGF can efficiently transfect BMSCs induced with 5-azacytidine, and this result provides basic experimental evidence for biotherapy of ischemic heart disease using BMSCs.
