水平振摇在骨髓间充质干细胞提纯中的作用

目的研究水平振摇对全骨髓贴壁分离大鼠骨髓间充质干细胞在细胞数量、纯度、凋亡及远期增殖及分化能力的影响,探索更为简单、高效的间充质干细胞提纯方法。方法在全骨髓贴壁法的基础上,使用水平摇床,以180 r/min、4 h为干预条件,水平振摇后计算贴壁细胞总数量;使用流式细胞仪测定贴壁细胞纯度及凋亡情况;使用MTT法测定振摇后贴壁细胞24 h的增殖情况;对分离纯化后细胞使用定向诱导培养基进行诱导分化。结果经振摇处理后,获得的细胞总量有所下降,细胞纯度显著提升,凋亡略有增加,增殖情况变化不大,具有向成骨细胞和脂肪细胞分化的能力。结论水平振摇法可显著提高全骨髓贴壁法的提纯效率,具有推广价值。

Exploration of horizontal shaking for bone marrow mesenchymal stem cells purification

Objective To study the effects of horizontal shaking on cell purity, activity, apoptosis, and differentiation in the process of the in vitro isolation and purification of rat bone marrow mesenchymal stem cells (BMSCs). Even the shaking effect for the cells proliferation, differentiation ability in the future. To establish a simpler and more efficient method for BMSCs purification. Methods Based on the whole bone marrow adhesion method, BMSCs isolated and purified by using different attachment method and shaking treatment of 180 r/min for 4 h, and then analyzed the adherent cell state including cell counting by cells counter; cell purity and apoptosis by flow cytometry; cells proliferation after shaking 24 h by MTT method; cells differentiation by directional induction medium. Results After shaking, cell count was decrease, purity was increased, apoptosis was increased, proliferation had no change, and the harvested cells had the ability to differentiate into osteoblasts and adipocytes. Conclusion The whole bone marrow adhesion method combined with appropriate shaking is more efficient and worth popularizing.