小鼠骨髓间充质干细胞的培养、鉴定和Survivin的表达研究

Culture,Identification of Mice Bone Marrow Mesenchymal Stem Cells and the Expression of Survivin

为培养及鉴定小鼠来源骨髓间充质干细胞,并测定细胞中Survivin的表达情况,采用全骨髓培养法获取骨髓间充质干细胞,绘制生长曲线,流式细胞仪检测细胞表面标志物,行成骨、成脂检测,RT-PCR测定Survivin表达情况.结果表明培养出的细胞呈长梭状成纤维细胞样,经流式细胞仪检测细胞表面高表达CD29、CD34、CD44、SCA-1,低表达CD117;细胞曲线显示传代细胞培养1～3d生长缓慢,第4d生长加快并于第7d达到高峰;成骨诱导20d经茜素红染色呈红色结节,成脂诱导14d油红O染色显示有大量脂质沉淀;RT-PCR结果显示Survivin mRNA阳性表达.经全骨髓培养法可以培养出大量骨髓间充质干细胞,同时Survivin在小鼠骨髓间充质干细胞中正常表达,提示可能参与骨髓间充质干细胞抗凋亡过程.

To culture and identify mice bone marrow mesenchymal stem cells （BMSCs）,and to detect the expression of Survivin,BMSCs by whole bone marrow adherence method was adopted,charted the growth curves of the passage BMSCs,the specific surface markers were detected by flow cytometry （FCM）,osteogenic and adipogenic induction was performed on BMSCs,expression of Survivin mRNA in BMSCs determined by RT-PCR.The results showed that the cells displayed fusiform shape as fibroblasts,the surface highly express CD29,CD34,CD44,SCA-1,lowly express CD117;the growth curve showed that cells grew slowly in the first three days,then growth accelerated and reached the top at the seventh day;after 20 days of osteogenic induction,alizarin red stain showed that alizarin red was positive in cells,and after 14 days adipogenic induction,oil red O stain showed that lipide sediment existed;Survivin mRNA was positive in BMSCs by RT-PCR.So BMSCs can be cultured abundantly by whole bone morrow adherence method,and expression of Survivin is positive,and indicating that Survivin may participate in the anti-apoptotic of BMSCs.