加压运动对羊骨髓间充质干细胞增殖、分化的影响

Effects of KAATSU Training on proliferation and differentiation of goat bone marrow mesenchymal stem cells

目的探索加压运动对羊骨髓间充质干细胞增值、分化的影响。方法将60只波尔山羊随机分成实验组和对照组,实验组每周进行2次加压运动,对照组非加压下运动,6个月后获取羊骨髓,然后采用percoll密度梯度离心法分离并吸取上中层界面间以单个核细胞为主的白色云雾层进行培养,观察细胞形态和增殖速率;两组细胞均用5-氮杂胞苷诱导其定向分化为心肌样细胞,对诱导后的细胞进行免疫荧光检测细胞上心肌特异性抗原a-actin表达情况。结果从骨髓中分离出的白色云雾层中的细胞能贴壁、能传代且生长状态良好。此外,经过5-氮杂胞苷诱导后细胞均表达心肌细胞特异性抗原。实验组骨髓间充质干细胞在培养过程中细胞多呈小而圆的形态,且增殖速率比对照组骨髓间充质干细胞快,而对照组细胞多呈多角形或梭形且增殖速率相对较慢。结论成功的分离并培养了骨髓间充质干细胞,并在体外成功诱导了骨髓间充质干细胞向心肌样细胞的分化。长期处于加压运动条件下的骨髓间充质干细胞比非加压运动条件下更倾向于增殖,而分化能力相对弱一些。

Objective To explore the effects of KAATSU Training on proliferation and differentiation of goat bone marrow mesenchymal stem cells.Methods 60 Boer goats were randomly divided into experimental group and control group,the experimental group were given KAATSU Training twice a week,non-KAATSU Training twice a week for the control group,6months later,we got the goat bone marrow,and then separated and absorbed the white cloud layer which mainly contained the mononuclear cell in the upper-middle interface with the method of percoll-density gradient centrifugation,cultured and observed the cell morphology and the proliferation rate;the cells of the two groups were induced into cardiomyocyte like cells by the 5-azacytidine,The cells which had been differentiated were detected the expression of the cardiac specific antigen a-actin by immunofluorescence assay.Results The cells isolated from the bone marrow in the white cloud layer were adherent,generated and grew well,in addition,the cells which induced by 5-azacytidine could express the cardiac specific antigen.The bone marrow mesenchymal stem cells of the experimental group were small and round,and their proliferation rate was faster than the control group’s,though the shape of the cells in the control group were polygonal or triangular,and the proliferation rate were slow.Conclusion It has been succeeded both in separation and cultivation of the bone marrow mesenchymal stem cells,and also induced the proliferation of turning into cardiomyocyte like cells in vitro.The bone marrow mesenchymal stem cells in pressurization motion for a long period of time were more easy to proliferate than the cells in non-pressurization motion,but the differentiative capability were low.