间充质干细胞培养中分化现象及c-fos表达研究

目的观察不同的培养基对间充质干细胞(MSC)体外自发分化的影响及c鄄fos的表达情况。方法以密度梯度离心法从SD大鼠骨髓中培养MSC,分别以L鄄DMEM、RPMI鄄1640与MSCGM作为培养基,观察对MSC在体外培养扩增的影响,流式细胞仪鉴定MSC,以免疫组化方法检测原癌基因c鄄fos的蛋白表达情况。结果MSCGM培养的MSC体外基本维持较均一的梭形,并能保持6代以上较少分化,而用L鄄DMEM和RPMI鄄1640培养,原代MSC形态基本较好,两代后开始大量分化。大部分MSC表达FOS蛋白。结论专用培养基对MSC体外培养中保持未分化状态很关键。FOS可能是MSC的增殖与分泌作用的胞内信号之一。

Differentiation and c-fos expression of mesenchymal stem cells in vitro

Objective To observe the self-differentiation of mesenchymal stem cells(MSC) when cultured in different culture medium and c-fos expressing of them in vitro. Method MSCs were isolated from SD rat's bone marrow by density gradient centrifugation and idenfied by FACS analysis. We used L-DMEM, RPMI-1640 and MSCGM, a special medium for MSCs, to observe the different effects and detected the c-fos expressing by immunohistochemical methods. Result The shape of MSCs were the same as shuttle in primary culture with three medium and it could maintain for 6 passages at least with MSCGM, but many of the MSCs self-differentiated in passage culture with L-DMEM and RPMI-1640.Most of the MSCs expressed c-fos protein.Conclusion It is very important for special culture medium to keep undifferentiation state in MSC culture. FOS is possible one part of the signal pathways for the proliferation and secretion of MSC.