大鼠骨髓间充质干细胞分离与培养:第3代细胞增殖快纯度高

发布时间：2018-01-04 22:10

本文关键词：大鼠骨髓间充质干细胞分离与培养:第3代细胞增殖快纯度高　出处：《中国组织工程研究》2014年50期 　论文类型：期刊论文

【摘要】：背景:进行组织工程研究的首要环节是培养出大量生物活性好、增殖速度快、细胞纯度高的种子细胞。目的:分析全骨髓贴壁法培养大鼠骨髓间充质干细胞的生物学特性。方法:采用全骨髓贴壁法分离、培养大鼠骨髓间充质干细胞,绘制第3代细胞生长曲线,观察各代细胞形态学变化,流式细胞分析法测定第2代、第3代细胞表面标志物(CD29、CD90、CD34、CD45)的表达。结果与结论:经全骨髓贴壁法培养3代以内骨髓间充质干细胞均具有典型成纤维样细胞形态而且贴壁生长;第2代骨髓间充质干细胞表型CD29、CD90、CD34、CD45表达分别为91.5%,96.2%,1.3%,8.0%,第3代骨髓间充质干细胞分别为98.2%,98.1%,0.6%,2.0%;细胞生长曲线结果表明,以细胞浓度8×107L-1接种,细胞增殖速度快,获得的细胞总数多。综合形态学及细胞鉴定结果,第3代大鼠骨髓间充质干细胞活性好、增殖速度快、细胞纯度高,适合作为组织工程研究的种子细胞。
[Abstract]:Background: the first step of tissue engineering research is to cultivate a large number of biological activity and rapid proliferation. Objective: to analyze the biological characteristics of rat bone marrow mesenchymal stem cells (BMSCs) cultured by whole bone marrow adherent method. Methods: rat bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured by whole bone marrow adherent method. The cell growth curve of the third generation was plotted and the morphological changes of each passage were observed. The surface markers of the second generation and the third generation were determined by flow cytometry (FCM). Results and conclusion: bone marrow mesenchymal stem cells (BMSCs) cultured by whole bone marrow adherent method all had typical fibroblast morphology and adherent growth. The phenotypic expression of CD29, CD90, CD34, CD45 in the second generation of bone marrow mesenchymal stem cells was 91.5% and 96.2%, respectively. The third generation of bone marrow mesenchymal stem cells were 98.2 and 98.1, respectively. The results of cell growth curve showed that when the cell concentration was 8 脳 10 ~ (7) L ~ (-1), the rate of cell proliferation was fast, and the total number of cells was more. The third generation of rat bone marrow mesenchymal stem cells have good activity, rapid proliferation and high purity, so they are suitable for tissue engineering research.
【作者单位】：成都医学院第一附属医院脊柱外科;中南大学湘雅医学院附属海口医院骨科中心;
【基金】：海口市重点科技项目(2011-0135,2014-082) 海口市人民医院院内课题(2014-YQY-07)~~
【分类号】：R329
【正文快照】： 0引言Introduction骨髓间充质干细胞是一类具有自我更新能力和多向分化潜能的细胞,具有取材方便、培养增殖速度快、无伦理学争议、低免疫原性等优点,目前已成为治疗脊髓损伤及其他神经系统疾病的良好供体细胞[1-7]。骨髓间充质干细胞是来源于骨髓组织的非造血干细胞,数量极少,

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