大鼠骨髓间充质干细胞的分离鉴定及免疫调节性能初探

发布时间：2018-01-05 02:33

本文关键词：大鼠骨髓间充质干细胞的分离鉴定及免疫调节性能初探　出处：《天津医药》2015年06期 　论文类型：期刊论文

【摘要】：目的分离培养大鼠骨髓间充质干细胞(BMSCs),鉴定其向脂肪细胞和软骨细胞的分化潜能,并探讨其免疫调节性能。方法无菌条件下摘取Sprague-Dawley(SD)大鼠股骨和胫骨,全骨髓贴壁法培养BMSCs,胰酶消化传代培养。待培养至第3代时进行流式鉴定,检测分离的BMSCs分化为脂肪细胞的潜能。与Wistar大鼠脾脏来源T细胞进行共培养,分为直接接触组和Transwell共培养组,探讨其对T细胞亚群的影响,并研究其相关机制。结果BMSCs呈梭形生长,生长至第3代的细胞经检测发现几乎不表达CD34和CD45,高表达CD29、CD44和CD90,且经诱导后细胞可分化为脂肪细胞和软骨细胞。与脾脏来源T细胞共培养可有效降低CD8+的效应T细胞(Teffs)的比例,并且增加CD4+CD25+双阳性的调节性T细胞(Tregs)数量,具有一定的免疫调节性能。与T细胞共培养后BMSCs的IL-10和TGF-β1基因表达显著增强,其中与T细胞直接接触组效果更为明显。结论 BMSCs可能通过直接接触以及分泌细胞因子来发挥免疫调节功能。
[Abstract]:Isolation and cultivation of rat bone marrow mesenchymal stem cells (BMSCs), to identify the adipogenic and chondrogenic differentiation potential, and to investigate its immunomodulatory properties. Methods under sterile conditions the removal of Sprague-Dawley (SD) of rat femur and tibia, whole bone marrow adherent culture BMSCs, trypsin digestion and flow passage. Type identification to be cultured to the third generation, detection and separation of BMSCs differentiation into adipocytes. Potential co cultured with Wistar rat spleen derived T cells, divided into direct contact co culture group and Transwell group, to investigate its effect on T cell subsets, and study its related mechanism. Results BMSCs was spindle growth, growth of the third generation cells were detected almost no expression of CD34 and CD45, the high expression of CD29, CD44 and CD90, and the induced cells can differentiate into adipocytes and chondrocytes. Co cultured with spleen derived T cells can effectively reduce CD8+. T cell (Teffs) ratio, and increased regulatory T cells CD4+CD25+ double positive (Tregs) number, regulate the performance of certain immune. T cells were co cultured with BMSCs IL-10 and TGF- beta 1 gene expression was significantly increased, and the T cell direct contact group effect is more obvious. Conclusion BMSCs may through direct contact and secretion of cytokines play a role in regulating immune function.

【作者单位】：天津医科大学基础医学院生物学教研室;
【基金】：国家高技术研究发展计划(863计划)资助项目(2012AA021003) 国家自然科学基金资助项目(21177091) 天津市科技支撑计划资助项目(12ZCZDSY03400)
【分类号】：R329.2
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