成人骨髓间充质干细胞的体外诱导培养及向脂肪细胞的定向分化

目的：确定人骨髓的间充质干细胞(MSCs)体外培养扩增、培养，向脂肪细胞表型转化的方法，并了解其生物学特性变化。方法：用Percoll分离液(1．073g／ml)分离成人MSCs，传代培养。流式细胞仪检测表面标记物CDl4，CD34，CD45，CD44，VLA—1，HLA—DR和细胞周期，用1—甲基—3—异丁基—黄嘌吟、吲哚美辛、牛胰岛素、地塞米松实现向脂肪细胞的定向诱导分化。油红O染色，光镜下观察橙红色脂滴沉着的细胞比例。结果：分离培养获得贴壁细胞，流式细胞仪检测CDl4，CD34，CD45，HLA—DR阴性，CD44阳性，VLA—1表达微弱。G0／G1期细胞约为86％。诱导72h后出现脂墒，第3周油红O染色示85％以上细胞转变为脂肪细胞。结论：从人骨髓分离、体外诱导培养间充质干细胞，可定向促进向脂肪细胞表型转化。

Isolation,culturation and adipogenisis committed differentiation of adult human mesenchymal stem cell

Objective: To isolate MSCs from adult human bone marrow cells and to induce them into adipocytes. Methods: MSCs were isolated from adult human bone marrow aspirated by Percoll and expanded in L DMEM. The surface antigen of MSCs, CD14, CD34, CD45, CD44, VLA 1, HLA DR and cell cycle were analysed on a FACScan flow cytometer. MSCs were cultured in adipogenisis inducing medium including insulin , 1 methyl 3 isobutylxanthine, indomethine and dexamethasone for 7 days and stained with Oil Red O. Results: MSCs grew as adherent cells and expanded more than 10 passages. They were positive for CD44 and negative for CD14, CD34, CD45, HLA DR. The expression of VLA 1 was weak. After 7 days of adipocyte inducing, about 85% of the cells displayed accumulation of lipid vacuoles, as detected by Red Oil O. Conclusion: MSCs isolated and cultured from adult human bone marrow can be induced to adipogenisis committed differentiation.