摘要
背景:细胞永生化是细胞获得持续增殖能力的一种特性,大T抗原基因可使细胞永生化效率提升,并广泛应用于实验中,越来越受到研究者的关注。目的:建立人骨髓间充质干细胞永生化细胞株,为人骨髓间充质干细胞的临床应用奠定基础。方法:使用含有猴空泡病毒40大T抗原基因的重组质粒载体转染人骨髓间充质干细胞,连续传代培养,通过形态学、细胞增殖能力、细胞表面标记、多向分化潜能等方法进行鉴定。结果与结论:①该方法构建的永生化细胞株为贴壁生长的梭形细胞,具有人骨髓间充质干细胞特异性表面标志;②转染后第50代人骨髓间充质干细胞仍然可表达猴空泡病毒40大T抗原基因;③永生化细胞株的增殖能力优于未转染人骨髓间充质干细胞;④永生化细胞株具有向成骨、成脂肪、成软骨细胞分化的潜能;⑤由此可见,转染猴空泡病毒40大T抗原基因不影响人骨髓间充质干细胞的生物学特性和分化能力,该方法可以用于建立人骨髓间充质干细胞永生化细胞株。
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.
作者
张露文
戴鹏秀
张翊华
陈奕静
王璟璐
李佳锴
Zhang Luwen;Dai Pengxiu;Zhang Yihua;Chen Yijing;Wang Jinglu;Li Jiakai(Northwest A&F University,Xianyang 712100,Shaanxi Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2022年第13期1969-1973,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(31872529),项目负责人:张翊华
陕西省自然科学基础研究计划重点项目(2019JZ-16),项目负责人:张翊华。
关键词
干细胞
人骨髓间充质干细胞
SV40大T抗原
基因
转染
永生化
鉴定
stem cells
human bone marrow mesenchymal stem cells
SV40 large T antigen
gene
transfection
immortalization
identification
