一种诱导神经干细胞向神经元定向分化的方法

A reliable method to induce neural stem cells directed differentiating into neurons in vitro

目的介绍一种诱导神经干细胞向神经元定向分化的方法。方法采用选择性无血清培养液培养大鼠神经干细胞。纯化培养2或3代后,将其接种于条件培养液中,诱导其向神经元定向分化。用倒置显微镜观察形态学变化,用NSE免疫细胞化学法检测其向神经元分化情况。结果条件培养液可有效诱导神经干细胞向神经元定向分化。将神经球消化成单细胞后接种,不但分化过程比神经球接种更同步,而且可提高其向神经元分化的比例。结论本方法可稳定高效地诱导神经干细胞向神经元定向分化。

Objective To introduce a reliable method to induce neural stem cells directed differentiating into neurons in vitro. Methods The rat neural stem ceils were cultured in selected serum-free medium. After cultured for 2, 3 passages, the neurospheres or single-cells isolated from neurospheres were cultured in conditioned medium to induce directed differentiating into neurons. Besides morphological observation of those cells under inverted microscope, NSE immunocytochemistry was carried out to detect the differentiating ratio of those cells from neural stem cells into neurons. Results It was shown that the conditioned medium could induce neural stem cells differentiating into neurons effectively in both neurospheres culture mode or single-cells culture mode. Furthermore, compared with the neurospheres culture mode, the single-cells culture mode showed more synchronous in the differentiation process and higher in the percentage of NSE-positive cells. Conclusion This method can induce neural stem cells directed differentiating into neurons effectively and stablely.