摘要
目的：探讨体外培养间充质干细胞的方法及其生物学特性，并检测与其抑制T淋巴细胞增殖活性减轻GVHD机制的细胞因子的表达。方法：通过密度梯度离心及贴壁筛选法分离、纯化及培养问充质干细胞，观察其生物学特性，并进行免疫细胞化学染色观察间充质干细胞是否表达TGF—β1、HGF。结果：采用贴壁筛选法培养间充质干细胞原代经7天贴壁后变为梭形，经14～17天可长满瓶底，传代后3～4天即可长满，免疫分型示CD29＋，CD34＋，CD34-，CD45-，免疫细胞化学染色显示高度表达TGF-β1及HGF。结论：密度剃度离心结合贴壁筛选法培养间充质干细胞可获得纯度较高，生物学特性稳定的问充质干细胞；间充质干细胞可能通过TGF-β1与HGF两种细胞因子抑制T淋巴细胞增殖减轻移植后GVHD。
Objective ： MSCs can express some cytokines to suppress T lymphocytes proliferation and alleviate GVHD. To investigate the biological characters of human bone marrow-derived mesenchymal stem cells（MSCs） in vitro and observe the cytokines expressed by MSCs. Methods ： Gradient centrifugation were used to separate, purify and culture MSCs, investigate the biological character of MSCs and observed if MSCs express TGF-β1 and HGF through immunochemistry. Results ： MSCs turned fibroblastic at 7 days after seeding and grow all of the culture bottom after 14 - 17 days culture. 3 - 4 days after passage can be passaged again. MSCs expressed CD29 and CD44, do not express CD34 or CD45, and through immunochemistry found TGF-β1 and HGF in MSCs were expressed highly. Conclusion ： By gradient centrifugation can gain purified MSCs with steady biological characters, hMSCs may suppress T lymphocytes proliferation through secreting TGF-β1 and HGF.
出处
《河南医学研究》
CAS
2006年第2期97-100,共4页
Henan Medical Research
