大鼠嗅鞘细胞无血清上清液诱导脊髓神经干细胞分化的研究

Study of the differentiation of NSCs induced by serum free condition medium of OECs from adult rat

[目的]观察大鼠嗅神经鞘细胞上清液对脊髓神经干细胞共培养发生的诱导分化作用。[方法]采用差速贴壁法获得较高纯度的嗅鞘细胞,分时段MTT法检测细胞活性,选取最佳状态细胞无血清培养后取上清液,与3代纯化后的神经干细胞共培养,观察分化过程,免疫荧光法鉴定诱导结果。[结果]MTT法分6个时段检测纯化后的嗅鞘细胞,发现9 d及12 d细胞活性最高。使用无血清嗅鞘细胞上清液与脊髓神经干细胞共培养发现诱导作用明显,向神经元样细胞及胶质细胞分化的比例分别达到53%和42%。[结论]嗅鞘细胞在体外培养的不同时间段活性不同,无血清的嗅鞘细胞上清有明显诱导神经干细胞向成熟神经元分化的作用。

[ Objective] AIM To explore the differentiation of neural stem cells from rat embryonic spinalcord in the serum free condition culture medium of OECs. [ Method] OECs were acquired by dissection ofolfactory nerve ensheathing and cultured for 20 hours, then suspending cells were put intoselective culture medium and cultured in the fibemectin coated plate. The activi- ty of OECs was detected by MTT method on different periods and the best situation was selected out and serum-free cultured. Exudates of OECs was added into neural stem cells of the third generation. The differentiation of neural stem cells were observed under inverted microscope. The neural stemcells cultured were identified by immumofluorescence method. [ Result]The activity of OECs was high on the 9＇h day and the 12＇h day. The serum free condition culture medium of OECs induced 53% neuralstem cells to differentiate into neuron-like cells and 42% neural stem ceils into astrocytes cells. [ Conclusion] The activity of OECs on different periods is different. The serum free condition culture medium of OECs can obviously induced neural stem cells to differentiate into mature neuronecells.