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目的观察人胰岛素样生长因子-1重组腺病毒(Ad-IGF-1)转染对C17.2神经干细胞体外增殖和分化的影响。方法体外培养及鉴定C17.2神经干细胞,Ad-IGF-转染C17.2神经干细胞;Western-blotting法检测IGF-1蛋白的表达,酶联免疫吸附实验(ELISA)检测IGF-1蛋白的表达曲线;MTT检测C17.2神经干细胞增殖,NSE及MBP免疫组化观察C17.2神经干细胞体外分化。结果转染AdIGF-1的C17.2神经干细胞成功表达IGF-1蛋白;IGF-1蛋白在病毒转染5～7d达高峰,13d仍高于转染前,Ad-IGF-1转染后各时间C17.2神经干细胞的增殖与对照组相比有显著差异,Ad-IGF-1转染对c17.2神经干细胞向神经元细胞及神经胶质细胞的分化与对照组相比有显著差异。结论Ad-IGF-1转染c17.2神经干细胞可稳定表达IGF-1,Ad-IGF-1转染促进C17.2神经干细胞的增殖及向神经元细胞和少突胶质细胞的分化。
Objective To observe the effects of recombinant human insulin-like growth factor-1 recombinant adenovirus (Ad-IGF-1) on the proliferation and differentiation of C17.2 neural stem cells in vitro. Methods C17.2 neural stem cells and Ad-IGF-transfected C17.2 neural stem cells were cultured and identified in vitro. IGF-1 protein expression was detected by Western-blotting and IGF-1 protein expression by enzyme-linked immunosorbent assay (ELISA) Curve; C17.2 neural stem cell proliferation, NSE and MBP immunohistochemistry were used to observe the differentiation of C17.2 neural stem cells in vitro. Results IGF-1 protein was successfully expressed on C17.2 neural stem cells transfected with AdIGF-1. The expression of IGF-1 protein peaked at 5-7 days after transfection and was still higher at 13 days than before transfection The proliferation of neural stem cells at time C17.2 was significantly different from that of the control group. Ad-IGF-1 transfected c17.2 neural stem cells differentiated into neurons and glial cells significantly compared with the control group. Conclusion Transfection of Ad-IGF-1 with c17.2 neural stem cells can stably express IGF-1. Ad-IGF-1 transfection can promote the proliferation of neural stem cells and the differentiation of neural stem cells and oligodendrocytes.