大鼠骨髓间充质干细胞分离鉴定方法及生物学特性

发布时间：2018-05-29 06:09

本文选题：骨髓 + 间质干细胞　；参考：《中国组织工程研究》2017年09期

【摘要】：背景:如何能简便高效地获得均质性的骨髓间充质干细胞群是软骨组织工程研究的重点。目的:分离培养大鼠骨髓间充质干细胞,观察其生物学特性。方法:抽取30只SD大鼠股骨以及胫骨骨髓,采用全骨髓贴壁培养法体外分离培养骨髓间充质干细胞,流式细胞仪测定细胞表面标记物的表达,细胞计数法观察细胞生长增殖能力,MTT法检测细胞存活率。结果与结论:(1)刚分离培养的细胞呈圆形、椭圆形,部分细胞呈三角形,接种12-15 d后,细胞开始融合,传代后细胞增殖速度加快,传代2 h后细胞均匀分布在瓶底;(2)随着传代次数的增加,细胞增殖能力出现下降趋势,第1-5代细胞存活率均超过95%,显著高于第6代和第7代(P0.05);(3)骨髓间充质干细胞表面CD34呈阴性表达,CD44呈阳性表达;(4)结果表明,采用全骨髓贴壁法可有效分离大鼠骨髓间充质干细胞,获得的细胞生长稳定,增殖能力强,可选用前5代细胞作为软骨组织工程的种子细胞。
[Abstract]:Background: how to obtain homogeneous bone marrow mesenchymal stem cells (BMSCs) is the focus of cartilage tissue engineering. Objective: to isolate and culture rat bone marrow mesenchymal stem cells (BMSCs) and observe their biological characteristics. Methods: bone marrow of femur and tibia of 30 Sprague-Dawley rats were extracted, bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adherent culture in vitro, and the expression of cell surface markers was measured by flow cytometry. Cell proliferation was observed by cell count method and cell survival rate was determined by MTT assay. Results and conclusion: the newly isolated and cultured cells were round, elliptical and triangular. After 12-15 days of inoculation, the cells began to fuse, and the proliferation rate of the cells increased after passage. After 2 h of passage, the cells distributed evenly at the bottom of the bottle.) with the increase of passage times, the proliferative ability of the cells decreased. The survival rate of the 1-5 passage cells exceeded 95%, which was significantly higher than that of the 6th and 7th generation (P0.05 + 3). The results showed that the bone marrow mesenchymal stem cells (BMSCs) could be effectively isolated by whole-bone marrow adherent method, and the positive expression of CD44 on bone marrow mesenchymal stem cells (BMSCs) was negative. The obtained cells can be used as seed cells for cartilage tissue engineering because of their stable growth and strong proliferative ability.
【作者单位】：江西医学高等专科学校;南昌大学医学部;
【分类号】：R318.08;R68

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