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目的建立生长因子定向诱导骨髓基质细胞(BMSCs)分化为胆碱能神经元的体系。方法 SD大鼠原代BMSCs体外培养,分别用神经生长因子(NGF)、NGF加碱性成纤维细胞生长因子(bFGF)、NGF加bFGF和表皮生长因子(EGF)诱导1、3、5 h,免疫荧光染色鉴定神经元表面标志微管联合蛋白-2(MAP-2)和胆碱能神经元标志物胆碱乙酰转移酶(ChAT),计数阳性细胞率。结果原代培养BMSCs传至第5代已基本纯化,NGF 100 ng/ml能有效诱导BMSCs分化为神经元,NGF、bFGF和EGF联合诱导可使BMSCs定向分化为胆碱能神经元,分化率为(67±8)%。结论 NGF、bFGF和EGF联合使用是诱导BMSCs定向分化为胆碱能神经元的有效组合,细胞分化率高。
OBJECTIVE: To establish a system in which growth factor-induced bone marrow stromal cells (BMSCs) differentiate into cholinergic neurons. Methods SD rat primary BMSCs were cultured in vitro and were induced by NGF, bFGF, bFGF and epidermal growth factor (EGF) for 1, 3, 5, Immunofluorescence staining identified neuronal surface marker microtubule-associated protein-2 (MAP-2) and cholinergic neuron marker choline acetyltransferase (ChAT), count the positive cell rate. Results Primary cultured BMSCs were purified to the fifth passage. NGF (100 ng / ml) effectively induced the differentiation of BMSCs into neurons. The combination of NGF, bFGF and EGF induced the differentiation of BMSCs into cholinergic neurons with a differentiation rate of (67 ± 8)%. Conclusion The combination of NGF, bFGF and EGF is an effective combination of inducing BMSCs to differentiate into cholinergic neurons with high cell differentiation rate.