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目的观察碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)联合诱导骨髓基质细胞(MSCs)能否分化形成神经球。方法采用全骨髓法培养MSCs,bFGF和EGF联合诱导MSCs,倒置显微镜下观察分化细胞的形态,免疫荧光法和免疫印迹法鉴定分化细胞。结果 bFGF和EGF联合应用,可有效地诱导MSCs形成神经球。此神经球能传代并分化成神经元样细胞。免疫荧光和免疫印迹法均证实神经球表达nestin蛋白,神经球分化的细胞表达神经元、胶质细胞和少突胶质细胞标志性蛋白。结论 bFGF和EGF联合应用,可有效地诱导MSCs形成神经球。此神经球能自我更新并分化为神经元、胶质细胞和少突胶质细胞。
Objective To observe whether bFGF and epidermal growth factor (EGF) induce the differentiation of bone marrow stromal cells (MSCs) into neurospheres. Methods MSCs were cultured by whole bone marrow method. MSCs were induced by combining bFGF and EGF. The morphology of differentiated cells was observed under inverted microscope. Differentiated cells were identified by immunofluorescence and Western blotting. Results The combination of bFGF and EGF could effectively induce the formation of neurospheres in MSCs. This neurosphere can be passaged and differentiated into neuron-like cells. Both immunofluorescence and immunoblotting confirmed that neurospheres express nestin, a neurosphere-differentiated cell that expresses neuronal, glial and oligodendrocyte marker proteins. Conclusions The combination of bFGF and EGF can effectively induce the formation of neurospheres in MSCs. The neurospheres self-renew and differentiate into neurons, glia and oligodendrocytes.