新医学2018，Vol.49Issue(5)：315-321,7.DOI:10.3969/j.issn.0253-9802.2018.05.004
体外诱导小鼠心脏CD51+细胞分化为血管内皮细胞的研究
In vitro induction of differentiation of cardiac CD51 +cells into vascular endothelial cells in mouse mod-els
摘要
Abstract
Objective To investigate the feasibility of isolation,culture and in vitro induction of the differentiation of cardiac CD5 1 +cells into vascular endothelial cells,aiming to provide theoretical evidence for the treatment of cardiac vascular endothelial injury. Methods The heart tissues were collected from 7-day-old C57 BL/6 mice. Primary cells were isolated by flow cytometry and cell sorting. The cardiac CD5 1 +cells were cultured and purified by suspension cell culture method. After cell purification and proliferation,the cardiac CD5 1 +cells were subjected to in vitro induction of differentiation into endothelial cells in endothelial cell growth medium-2(EGM-2)for 20 days. After in vitro induction,the changes in cell morphology were ob-served. The expression levels of CD31,CD34,vWF,VEGFR-2 and VE-Cadherin mRNA were quantitatively measured by real-time fluorescent quantitative PCR. The expression levels of CD31,VEGFR-2 and vWF were detected by immunofluorescent staining and flow cytometric immunophenotying. The level of nitric oxide(NO) was measured and the function of the induced endothelial cells was evaluated by angiogenesis assay in vitro. Results The morphology of cardiac CD5 1 +cells was changed from the long-spindle to short-spindle shape and the induced cells grew in a cobblestone-like pattern. The results of fluorescent quantitative PCR demonstrated that the relative expression levels of CD31,CD34,vWF,VEGFR-2 and VE-Cadherin mRNA were significantly up-regulated after induction(t=24.529,6.383,25.872,5.256 and 5.255,all P<0.05). Both immunoflu-orescent staining and flow cytometric immunophenotying revealed that the cells expressed CD31,VEGFR-2,and vWF after induction. The quantity of NO was considerably increased after induction(t =9.549,P=0.011)and had the ability to form lumen-like structures in vitro. Conclusion Cardiac CD51 +cells can be induced to differentiate into vascular endothelial cells in vitro.关键词
间充质干细胞/CD51/内皮细胞/诱导分化Key words
Mesenchymal stem cell/CD51/Endothelial cell/Induced differentiation引用本文复制引用
高山泉,廖延,姜美花,杜钰,谢冬梅,祝双华,彭朝权..体外诱导小鼠心脏CD51+细胞分化为血管内皮细胞的研究[J].新医学,2018,49(5):315-321,7.基金项目
国家自然基金面上项目(81370214),广东省自然科学基金面上项目(S2013010016392),广东省自然科学基金自由申请项目(2015A030313183) （81370214）
