共培养体系间充质干细胞对同源内皮祖细胞增殖和血管形成的影响  

Mesenchymal stem cells promote the proliferation and angiogensis of endothelial progenitor cells in the co-culture system

作　　者：冯文磊 张猛[1] 徐芳洁[2] 王艳杰[1] 陈雪玲[2] 吴向未[1] 

机构地区：[1]石河子大学医学院第一附属医院普外科,石河子医学硕士研究生832008 [2]石河子大学医学院免疫学教研室,石河子832002

出　　处：《医学研究生学报》2015年第3期229-235,共7页Journal of Medical Postgraduates

基　　金：国家自然科学基金(31271458);人力资源和社会保障部留学回国人员科技活动项目(RSLX201201);新疆建设兵团科技计划项目(2014AB047)

摘　　要：目的临床干细胞治疗心肌梗死可能归因于细胞旁分泌对细胞存活和血管形成的作用,文中探讨小鼠骨髓来源的间充质干细胞(mesenchymal stem cells,MSCs)在非接触共培养体系中对同源内皮祖细胞(endothelial progenitor cells,EPCs)增殖和血管形成的影响。方法小鼠骨髓细胞经差速贴壁法分离MSCs与EPCs,以流式细胞术(flow cytometry,FCM)检测MSCs和EPCs表面标记物,以成骨成脂成软骨诱导分化对MSCs进行功能鉴定,以成血管对EPCs进行功能鉴定。取第3代MSCs和EPCs,按1∶1的细胞比例接种入Transwell共培养系统中,以下室接种EPCs上室接种MSCs为实验组,同时设置相同密度单纯EPCs接种于下室为对照组。采用MTT比色法和Ed U荧光标记法检测MSCs对EPCs增殖能力的影响,采用计数血管样结构数目和联结点数目检测MSCs对EPCs血管形成能力的影响。结果第3代MSCs经FCM检测高表达Sca-1,CD29低表达CD45,CD11b;经诱导可向成骨成软骨成脂方向分化。第3代EPCs经FCM检测高表达CD34、CD133和VEGFR2,在基质胶上可形成血管样结构。MTT比色法和Ed U荧光标记法结果显示与对照组相比,实验组细胞的增殖能力在72 h后显著增强(P<0.05);实验组DNA合成期的细胞比例较对照组显著增多[(89.00±5.30)vs(76.00±3.38),P<0.01)]。倒置显微镜观察结果显示实验组EPCs生成的血管样结构和联结点数目较对照组明显增加[(13.53±2.33)vs(10.80±1.82)、(16.60±2.23)vs(13.07±2.81),P<0.05]。结论利用差速贴壁法可同时分离纯化扩增MSCs和EPCs。EPCs在和MSCs非接触共培养时其增殖能力和血管形成能力都得到提高。Objective Stem cell-based therapy can achieve several functional improvements in the treatment of myocardial infarction, which are reportedly attributed to its paracrine effect on cell survival and angiogenesis .This study was to isolate and co-cul-ture mesenchymal stem cells （MSCs） and endothelial progenitor cells （EPCs） from the bone marrow of C57B/L6 mice in non-contact co-culture systems to demonstrate the contribution of MSCs to the proliferation and angiogenesis of EPCs . Methods EPCs and MSCs were isolated from the bone marrow of C57B/L6 mice using the differential adhesion method and their surface markers were identified by flow cytometry .Functional identification of the MSCs was performed by osteogenesis-, adipogenesis-and chondrogenesis-inducing differentiation , and that of the EPCs by angiogenesis experiment .The passage-3 EPCs and MSCs were harvested and placed into the Tr-

关 键 词：内皮祖细胞 间充质干细胞 非接触共培养 增殖 血管形成 

分 类 号：R541[医药卫生—心血管疾病]