不同pH值的培养液对兔骨髓间充质干细胞体外培养的影响

The influence of different pH level of DMEM on rabbit Bone Mesenchymal stem cells in vitro culture

目的 探讨不同pH值的低糖DMEM培养液对兔骨髓间充质干细胞 (MSCs)体外培养的影响 ,寻找最适宜MSCs生长的培养液的pH值。方法 采用常规的细胞培养传代技术以及光、电镜技术和细胞增殖活性的检查方法 ,观察不同pH值的低糖DMEM培养液对MSCs生长、增殖、形态的影响。结果 培养液的pH值在 7.2～ 7.4之间时 ,MSCs生长活跃 ,增殖速度极快 ,无接触抑制 ,镜下细胞呈单一梭形外观的成纤维细胞 ,形成成纤维细胞集落。pH值大于 7.4时 ,MSCs逐渐停止增殖 ,镜下细胞膜贴壁展开、边缘卷曲 ,细胞面积变大为正常的数倍 ,形态极不规则。pH值在 7.1～ 7.19之间时 ,变化不大 ,仅增殖速度稍减慢。pH值小于7.1时 ,MSCs生长增殖速度亦明显减慢 ,镜下细胞逐渐从培养瓶壁脱落 ,成为圆球形悬浮细胞。结论 MSCs对碱性的培养液较敏感 ,稍偏碱即影响生长增殖 ,对酸性的培养液则有一定的耐受。使用培养液前有条件也最好用pH计测定pH值 ,同时注意观察细胞生长增殖情况及培养瓶中培养液的颜色 。

Objective To study the influence of different pH level of DMEM (Dulbecco's modified Eagle's medium) on rabbit Bone Mesenchymal stem cells in vitro culture to find the optimum pH level. Methods The influence of different pH level of DMEM on MSCs in vitro culture were studied by the normal way. The growth, the proliferation and the morphology were observed. Results The growth and the proliferation velocity of MSCs were active and fast respectively when the pH level of culture medium were between 7.2～7.4, MSCs formed the cell clone having the anagogic cell shape and exhibited a typical fibroblast-like morphology. The situation was similar to former when the pH level of culture medium were between 7.1～7.19. But when the pH level of culture medium was over 7.4,the proliferation velocity of MSCs obviously declined, and the elicited outspread , fringe curl , and the cell shape were abnormal. The proliferation velocity of MSCs also obviously declined when the pH level of culture medium was less than7.1. MSCs gradually desquamated from the wall and became a ball-like appearance. Conclusion MSCs were sensitive to the alkalescent culture medium, but it could bear acidic solution in a certain extent. Culture medium must be measured of pH level before using it. In order to replacing the fresh culture medium in time, the situation of proliferation and the color of culture medium should be observed carefully.