CXCR4基因转染的猪BMSCs体外诱导分化为心肌样细胞的实验研究.docVIP

CXCR4基因转染的猪BMSCs体外诱导分化为心肌样细胞的实验研究 　　作者：陈中璞1，李拥军2 ，姚玉宇2，蒋益波1，钱琪1，潘啸东1 作者单位：(1. 东南大学医学院,江苏 南京 210009; 2. 东南大学附属中大医院 心内科,江苏 南京 210009) 　　【摘要】 目的:探讨基质细胞衍生因子1(SDF1)受体CXCR4基因转染的猪骨髓间充质干细胞(BMSCs)体外诱导分化为心肌样细胞的可行性。方法：按照Nance方法培养BMSCs，转染带荧光的CXCR4基因，流式细胞仪检测转染后BMSCs的细胞周期，用5氮胞苷(5aza)诱导，并行结蛋白、肌球蛋白重链、心肌特异性肌钙蛋白I免疫组化鉴定。对照组为未转染CXCR4基因的BMSCs。结果：体外培养的原代BMSCs 10～14 d达到融合，CXCR4基因转染后的BMSCs能成功表达CXCR4，转染前后细胞周期无明显改变。与对照组相同，经5aza刺激后，部分BMSCs成梭形，结蛋白、肌球蛋白重链、心肌特异性肌钙蛋白I免疫组化染色均为阳性。结论:CXCR4基因转染的猪BMSCs在体外条件下生长稳定,传代后仍保持未分化状态,有分化成心肌样细胞的潜能。 　　【关键词】 基质细胞衍生因子1; CXCR4基因; 骨髓间充质干细胞; 体外诱导; 心肌样细胞 　　Investigation on inducing porcine BMSCs transfected by CXCR4 gene 　　into cardiomyocyteslike cells in vitro 　　CHEN Zhongpu1， LI Yongjun2， YAO Yuyu2， JIANG Yibo1， QIAN Qi1, PAN Xiaodong1 (1. Medical College, Southeast University, Nanjing 210009, China; 2. Department of Cardiology, Zhongda Hospital, Southeast University, Nanjing 210009, China) 　　[Abstract] Objective: To investigate the possibility of inducing porcine bone marrow mesenchymal stem cells(BMSCs) which were transfected by CXCR4 gene first,the acceptor of SDF1, to differentiate into cardiomyocyteslike cells in vitro. Methods: BMSCs were isolated from bone marrow and purified by centrifuge.The proliferation and growth characteristics were observed in primary and passage culture.BMSCs were transfected by CXCR4 gene, then analyzed cell cycle by flowcytometer.After being cocultured with 5aza cytidine for 24 h, the cultured cells were evaluated by immunohistochemical stains. The control group was BMSCs without CXCR4 gene. Results: The cells were fused in single layer after plating for 10～14 days.BMSCs which were transfected could express CXCR4 successfully, and the cell cycle didnt changed. The same as the control group,after being cocultured with 5aza cytidine, some of transfected BMSCs became spindlelike and formed a network of myotubule. Myotubular cells were found to be stained with positively MHC, desmin and TnI. Co