窦房结细胞裂解液对大鼠骨髓间充质干细胞的诱导分化作用

摘要

目的:观察不同浓度的窦房结细胞裂解液对骨髓间充质干细胞(MSCs)的诱导分化作用.方法:体外分离乳鼠窦房结细胞并制备窦房结细胞裂解液.密度梯度离心法分离成年SD大鼠骨髓MSCs,分别加入含1、2、4和8倍浓度的窦房结细胞裂解液培养基诱导培养,普通培养基培养作对照,观察细胞形态变化,并通过免疫荧光染色法检测MSCs心肌特异性肌钙蛋白I(cTnI)的表达,免疫细胞化学染色检测超极化激活的环核苷酸门控的离子通道蛋白HCN2和HCN4的表达.结果:经1、2、4和8倍浓度窦房结细胞裂解液诱导1周后的骨髓MSCs均表达cTnI和HCN2;5组间HCN2阳性细胞率比较,差异有统计学意义(F=93.113,P＜0.001),且在一定浓度下随着裂解液浓度的升高而增高(P＜0.05).结论:窦房结细胞裂解液能够体外诱导骨髓MSCs分化为具有起搏功能的心肌样细胞.%To observe the effects of different concentration of lysate of sinoatrial cells on differentiation of the rat bone marrow mesynehymal stem cells( MSCs). Methods:Sinoatrial cells of rats were extracted in vitro and then were made into lysate. Dissociated the rat MSCs and induced them with different concentration of lysate of the sinoatrial cells. The group with ordinary medium was chosen as control group. The morphological changes were observed, and the expressing of cardiac cell specific protein cTnI,HCN2 ,and HCN4 were analyzed by immunofluoresence and immunohistochemistry. Results: Different concentration of lysate of the sinoatrial cells induced the MSCs to differentiate into cardiac cells, which expressing HCN2. To some extent, the higher the concentration of lysate of the sinoatrial cells was, the higher rates of the speed of differentiation ( F = 93. 113,P <0.001). Conclusion; Lysate of the sinoatrial cells could induce the MSCs to differentiate into cardiac cells with pacemaking functionin in vitro, which establishes theoretical foundation for the choice of seed cells on biological pacemaker.