大鼠骨髓间充质干细胞的分离鉴定及免疫调节性能初探  

Isolation of rat bone marrow-derived mesenchymal stem cells and explore its role in immunomodulation

作　　者：张志强[1] 刘义[1] 李克秋[1] 李光[1] 

机构地区：[1]天津医科大学基础医学院生物学教研室,300070

出　　处：《天津医药》2015年第6期611-615,共5页Tianjin Medical Journal

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)

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。Objective To explore the immunomodulation property of bone marrow-derived mesenchymal stem cells （BMSCs） from Sprague-Dawley （SD） rats after they are isolated, cultured and identified by surface marker and differentiation potential examination. Methods BMSCs were isolated from femur and tibia of SD rats and passaged by trypsinization. The surface markers of the 3rd passage BMSCs were detected by flow cytometry and the capacity of their adipocyte and cartilage differentiation were examined. In order to explore the immunomodulation property of BMSCs, allogeneic spleen T cells of Wi-star rats were co-cultured with BMSCs through either cell-to-cell contact or transwell, then its effect on the T cell subsets and related mechanism was also examined. Results BMSCs were mainly spindle-shaped in culture. Surface marker detec-tion showed that BMSCs expressed high levels of CD29, CD44 and CD90 but no CD34 nor CD45 at the third generation. Un-der specific condition, BMSCs could differentiate into adipocytes and chondrocytes. The CD8＋effector T cells （Teffs） decreas-es effectively and the CD4＋CD25＋regulatory T cells （Tregs） increased remarkably when BMSCs were co-cultured with allo-geneic spleen T cells for 48 hours. The expressions of IL-10 and TGF-β1 of BMSCs significantly increased after co-culture with T cells, and this effect was more obvious in cell-to-cell contact group. Conclusion The immunomodulation property of BMSCs were presumably function through cell-to-cell contacts and cytokine secretion.

关 键 词：骨髓 间质干细胞 细胞分化 神经免疫调节 大鼠 Sprague-Dawley 大鼠 Wistar 

分 类 号：R349.5[医药卫生—基础医学]