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2022, Vol. 26 ›› Issue (13): 1969-1973SV40 large T gene induces immortalization of human bone marrow mesenchymal stem cellsZhang Luwen, Dai Pengxiu, Zhang Yihua, Chen Yijing, Wang Jinglu, Li Jiakai
Abstract: BACKGROUND: Cell immortalization is a characteristic of cells acquiring the ability to continue to proliferate. The large T antigen gene can improve the efficiency of cell immortalization, and is widely used in experiments, attracting more and more attention from researchers. OBJECTIVE: To establish an immortalized cell line of human bone marrow mesenchymal stem cells and lay the foundation for the clinical application of human bone marrow mesenchymal stem cells. METHODS: The recombinant plasmid vector containing the SV40 large T antigen gene was used to transfect human bone marrow mesenchymal stem cells, which were successively subcultured and identified by methods, such as morphology, cell proliferation ability, cell surface markers, and multidirectional differentiation potential. RESULTS AND CONCLUSION: (1) The immortalized cell line constructed by this method is an adherent growth of spindle cells with specific surface markers of human bone marrow mesenchymal stem cells. (2) The 50th generation of human bone marrow mesenchymal stem cells can still express SV40 large T antigen gene after transfection. (3) The proliferation ability of the immortalized cell line is better than that of untransfected human bone marrow mesenchymal stem cells. (4) The immortalized cell line has the potential to differentiate into osteogenic, adipogenic, and chondrogenic cells. (5) This shows that transfection of the SV40 large T antigen gene does not affect the biological characteristics and differentiation ability of human bone marrow mesenchymal stem cells. This method can be used to establish immortalized human bone marrow mesenchymal stem cells. Key words:stem cells, human bone marrow mesenchymal stem cells, SV40 large T antigen, gene, transfection, immortalization, identification |