[Differentiation of 5-azacytidine-induced bone marrow stromal stem cells transduced with hepatocyte growth factor gene].
OBJECTIVE
To 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.


METHODS
5-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.


RESULTS
After 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.


CONCLUSIONS
Ad-HGF can efficiently transfect BMSCs induced with 5-azacytidine, and this result provides basic experimental evidence for biotherapy of ischemic heart disease using BMSCs.
Wen
Qian
Q
Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China.
Zhou
Chao-ying
CY
Zhou
Ming-qian
MQ
Luo
Wei
W
Ma
Li
L
D000818:Animals; D001374:Azacitidine / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D004285:Dogs; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000235:genetics* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology; D014162:Transfection
D017228:Hepatocyte Growth Factor; D001374:Azacitidine