人骨髓间充质干细胞表面抗原测定

目的 :研究体外培养的各代间充质干细胞 (MSCs)及其成骨分化后的部分细胞表面抗原变化。方法 :将体外普通培养的骨髓间充质干细胞 ,每隔 1代在流式细胞仪上检测CD48、CD5 6、CD71和CD90阳性表达率。另外 ,为促进成人MSCs体外成骨性分化 ,第 1传代培养时加入成骨性添加剂 ,培养第 6、 12、 18、 2 4、 3 0d时 ,也分别用流式细胞仪分析上述MSCs表面抗原的表达 ,并用碱性磷酸酶组化染色和VonKossa染色。结果 :( 1)在普通培养条件下 ( 10 %DMEM ) ,从第 1代起 ,MSC的表面抗原CD48、CD5 6、CD71、CD90都呈阳性表达。 ( 2 )诱导培养的MSCs表面抗原CD48、CD90阳性表达率在诱导第 12、 18、 2 4及 3 0d时 ,分别与第 6d相比都有显著差异。 ( 3 )在成骨诱导条件下 ,MSCs表现出AKP染色增强和VonKossa染色可见钙化的基质沉积。当MSC开始向成骨细胞分化后 ,CD71阳性率下降最明显。结论 :( 1)目前采用的体外普通培养条件下 ,培养、传代方法对MSCs表面抗原CD48,CD5 6,CD71及CD90表达并无明显影响。 ( 2 )诱导成骨培养的MSCs的CD71从高阳性率转变为极低阳性率可能与MSCs向成骨细胞系的分化成熟相对应。 ( 3 )CD90及CD48也可能与MSCs的细胞分化状态特别是向成骨细胞系的改变有关

Detection of cell surface molecules on adult MSCs cultured in vitro

Objective:To explore the possibility of 5-azacytidine(5-Aza) inducing mesenchymal stem cells (MSCs) differentiating into myoblast(Skeletal Muscle ) in vitro.Method:Cell surface molecules CD48,CD56,CD71 and CD90 on the first,third,fifth and seventh passage MSCs were analyzed by flow cytometry technique.For in vitro osteogenic differentiation,adult hMSCs from the first passage were grown in the presence of osteogenic supplements.On days 6~(th),12~(th),18~(th),24~(th) and 30~(th) of culture,flow cytometic analysis was performed to examine the expression of cell surface molecules CD48,CD56,CD71 and CD90 on adult hMSCs and samples were assayed for alkaline phosphatase histochemistry and for mineral deposition by Von Kossa staining respectively.Result:(1) The adult hMSCs express CD48,CD56,CD71 and CD90.(2) When the cells were induced by OS medium,there was significant difference referred to the positive rate of CD48,CD71 and CD90 on days 12~(th),18~(th),24~(th) and 30~(th) compared with that on day 6~(th).(3)Under the function of osteogenic supplements,the adult hMSCs were found higher activity of alkaline phosphatase,and depositing a calcified matrix.At the same time,the positive rate of CD71 decreased obviously.Conclusion:(1) The present method we used to isolate and culture the adult hMSCs has no obviously influence on the expression of CD48,CD56,CD71 and CD90.(2)There is correlation between the positive rate of CD71 and the osteogenesis of adult hMSCs.(3)There also might be relationship between the rate of CD90 and CD48 and the differentiation of adult hMSCs.