大鼠骨髓间充质干细胞诱导分化为表皮细胞的实验观察

Study on induction of mesenchymal stem cells into epidermal cells in rat

目的:研究表皮生长因子(Epidermal growth factor,EGF)加条件培养基体外诱导大鼠 MSCs 向表皮细胞定向分化的可行性。方法:采用 Ficoll-Paque 淋巴细胞分离液分离扩增大鼠骨髓 MSCs,免疫细胞化学染色及流式细胞仪进行鉴定。传至第3代的大鼠 MSCs 用表皮生长因子(EGF)、条件培养基等定向诱导 MSCs 分化为表皮细胞;免疫细胞化学对细胞角蛋白 CK5/8、19(Cytokeratin5/8、19)阳性表达细胞进行检测。结果:从大鼠骨髓中分离培养的 MSCs 增殖能力强,细胞表面标志 CD34、CD45阴性,CD29、CD44阳性,流式细胞仪检测显示细胞纯度高,诱导后7d 细胞免疫化学显示角蛋白5/8、19染色阳性,具有表皮细胞特征。结论:从大鼠骨髓中分离培养出的问充质干细胞,具有自我更新和增殖能力强的特点,经诱导可定向分化表达角蛋白。

Objective:To investigate the feasibility of inducing bone marrow mesenchymal stem cells(MSCs) into epidermal cells with epidermal growth factor (EGF) and conditioned culture medium in vitro.Methods:Bone mar- row MSCs were isolated from rat bone marrow using Ficoll-Hypague lymphocyte separation medium,and they were proliferated in vitro.Immunocytochemistry and flow cytometric examinations were carried out to identify the MSC cells.MSCs were then cultured in a conditioned culture medium to induce the transformation of epidermal cells,which were identified with cytokeratin5/8 and cytokeratin19 expression.Results:Bone marrow MSCs prolif- erated quickly and were uniformly positive for CD29 and CD44 but negative for CD34 and CD45.After being in- duced for seven days with conditioned culture medium,they manifested the characteristics of epidermal cells as evi- denced by cytokeratin5/8 and 19 expression.Conclusion:MSCs derived from rat proliferated rapidly and could be induced into epidermal cells.