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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 epidermal cells induced by epidermal cells induced by epidermal growth factor (EGF) plus conditioned media in vitro. Methods: Bone marrow MSCs were separated and amplified by Ficoll-Paque lymphocyte separation and identification by immunocytochemistry and flow cytometry. MSCs transfected into passage 3 were induced to differentiate into epidermal cells with epidermal growth factor (EGF) and conditioned medium. Immunocytochemistry was used to detect the expression of cytokeratin 5/8 and 19 Cells are tested. Results: The MSCs isolated from rat bone marrow showed strong proliferative ability. The cell surface markers CD34 and CD45 were negative, CD29 and CD44 were positive. The purity of the cells was high by flow cytometry. Immunocytochemistry showed that keratin 5/8 , 19 stained positive, with epidermal cell features. CONCLUSION: The mesenchymal stem cells isolated from the bone marrow of rats possess the characteristics of self-renewal and proliferation. They can be induced to differentiate and express keratin.