猪骨髓间充质干细胞的分离培养及体外转化为心肌样细胞的实验研究  

Culture of multipotent mesenchymal stem cells from porcine bone marrow and transformation to myogenic cell.

作　　者：李厚怀[1] 沈振亚[1] 滕晓梅[1] 郑琳[1] 李科[1] 焦朋[1] 周晓彤[1] 夏漫辉[1] 

机构地区：[1]苏州大学附属第一医院,在读博士生江苏苏州215006

出　　处：《山东医药》2005年第6期12-13,共2页Shandong Medical Journal

基　　金：江苏省卫生厅 13 5工程资助课题 (No.RC2 0 0 3 0 92 )

摘　　要：目的　探讨体外培养猪骨髓间充质干细胞 (MSCs)的方法及 MSCs的一些生物学特性 ,为细胞心肌成形术提供新材料。方法　按照 Nancer方法培养 MSCs,用 5 -氮胞苷 (5 - aza)诱导 ,流式细胞仪检测细胞周期 ,并行结蛋白 (desmin)、肌球蛋白重链 (MHC)、心肌特异性肌钙蛋白 I(c Tn I)、连接蛋白 - 4 3(Cx- 4 3)免疫组化鉴定。结果　体外培养的原代 MSCs10～ 14天达到融合 ,细胞周期显示 80 %的细胞处于 G0 / G1期 ;经 5 - aza刺激后 ,部分MSCs呈梭形 ,结蛋白、肌球蛋白重链、心肌特异性肌钙蛋白 、连接蛋白 - 4 3免疫组化染色阳性。结论　猪 MSCs在体外条件下生长稳定 ,传代后仍保持未分化状态 ,有分化成心肌的潜能 ,可作为细胞心肌成形术的材料。Objective By establishing a method for the culture of porcine mesenchymal stem cells(MSCs) from bone marrow and the transformation to myogenic cells ex vivo,to provide a new cell source for the cellular cardiomyoplasty.Methods MSCs were isolated from bone marrow and purified by centrifuge.The proliferation and growth characteristics were observed in primary and passage culture.Cell cycle was analyzed by measuring DNA content with flowcytometer.After being co-cultured with 5-aza cytidine for 24h,the cultured cells were evaluated by immunohistochemical stains.Results The adherent,fibroblast-like cells were confluent in single layer after plating for 10～12 days.The cell cycle analysis showed that 80% of MSCs were in G0/G1 phase.After being co-cultured with 5-aza cytidine,some of MSCs became spindle-like and formed a network of myotu- bules.Myotubularcells were found to be stained with positively MHC,cTnI,Cx-43 and desmin.Conclusion Porcine MSCs can be isolated from postnatal bone marrow through their adherentability.Myogenic cells can be generated from MSCs in vitro MSCc may be a new cell source for the cellar cardiomyoplasty.

关 键 词：细胞培养 猪 骨髓间充质干细胞 细胞分离 体外转化 心肌样细胞 细胞心肌成形术 

分 类 号：R654.2[医药卫生—外科学] R329.2[医药卫生—临床医学]