鼠胶质瘤细胞上清液诱导人骨髓间充质干细胞向神经元样细胞的分化

Differentiation of bone mesenchymal stem cells into neuron-like cells induced by supernatant of glioma cells

目的:分离培养人骨髓间充质干细胞(MSCs),探讨鼠胶质瘤细胞上清液对其向神经元样细胞的诱导分化。方法:利用Percoll梯度分离法,培养人MSCs。采用鼠胶质瘤细胞上清液对MSCs进行诱导分化。观察人MSCs经诱导后细胞的形态变化,采用免疫细胞化学方法检测神经元烯醇化酶(NSE)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)的表达。结果:诱导24h后,细胞胞体收缩呈锥形或球形,有突起长出,细胞间突起相互连接,交错成网,为典型的神经元样形态。免疫细胞化学结果显示,经诱导培养后的神经元样细胞的胞体及部分突起NSE和NF染色呈强阳性表达,而GFAP染色呈阴性。诱导组出现NSE阳性的细胞率为(79.5±3.2)%,而对照组出现NSE阳性的细胞率为(12.1±2.0)%,两组之间比较差异有显著性(P<0.01)。诱导组出现NF阳性的细胞率为(41.2±2.4)%,而对照组为阴性。结论:鼠胶质瘤细胞上清液可以诱导MSCs向神经元样细胞分化。

Objective To study the isolation, cultivation of human mesenchymal stem cells （MSCs） and the differentiation into neuron-like cells induced by the supernatant of glioma cells in vitro. Methods MSCs were separated from human bone marrow and expanded in vitro and induced by the supernatant of glioma cells. The morphological changes of MSCs after induction were obeserved. The expressions of neuron specific enolase （NSE）, neurofilm entprotein （NF） and glial fibrillary acidic protein （GFAP） were detected by immunohistochemistry. Results After induction for 24 h, the differentiated cells showed typical neuronal appearance, and expressed neuron-specific markers, such as NF and NSE. But glial astrocyte marker GFAP was negative. Quantitative count analysis showed that the NSE positive rate of the cells in the induced group （79.5 % ±3.2 %） was much higher than that in the control group （12. 1%± 2.0%, P〈0. 01） . The NF positive rate of cells in the induced group was 41.2 % ±2. 4%, and the cells in the control group did not express NF. Conclusion The supernatant of glioma cells has inductive effect on differentiation of MSCs into neuron-like cells in vitro.