CONSTRUCTION OF IMMORTALIZEDBONE MARROW MESENCHYMAL STEM CELLS / 现代医院

ABSTRACT

Objective To establish immortalized bone marrow mesenchymal stem cells (BMSCs) from SD rats in vitro for further research on the characteristics and clinical application of BMSCs.Methods By using L ipo-fectamine TM 2000 -mediated gene transfection, plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into BMSCs.BMSCs were screened by puromycin and then cultured on an extended scale.Their cell morphology and growth conditions were observed.Growth curve of cells was graphed.The expression of SV40Tag in transfected cells was identified by enzyme digestion method for the detection of tumor forma-tion.Results The growth rate of the experimental group of BMSCs after the 7th generation was significantly higher than that of the control group.No tumorigenicity was found in BMSCs after the 7th generation.Conclusion In vitro, the immortalized BMSCs by pCMVSV40T/PUR can provide basis on a large scale for its application in clinic and sci-entific research.