条件培养基对神经干细胞增殖与分化作用的影响

目的：观察大鼠骨髓间充质干细胞（BMSCs）条件培养基对体外培养神经干细胞（NSCs）增殖与分化作用的影响。方法用 BMSCs 条件培养基分别在增殖与分化条件下培养 NSCs，观察 NSCs 形态变化。用 MTT 法及 NSCs 球数目和直径的统计分析了解 NSCs 增殖情况，同时行免疫荧光及 Western blot 法鉴定其分化情况。结果原代培养获得大量未分化且悬浮生长的 NSCs 球，并能分化为神经元细胞、星形胶质细胞及少突胶质细胞。BMSCs 条件培养基组能够促进 NSCs 球的吸附贴壁，但与对照组相比光密度（OD）值差异无统计学意义，NSCs 球数目和直径差异亦无统计学意义。另外，BMSCs 条件培养基组少突胶质细胞特异性蛋白MBP 表达量高于对照组（P <0.05），星形胶质细胞特异性蛋白 GFAP 表达量低于对照组（P <0.05），而神经元特异性蛋白 MAP-2表达量未见明显异常。结论大鼠 BMSCs 条件培养基促进 NSCs 向少突胶质细胞方向分化，但不抑制其增殖作用。

Effects of conditioned medium on the proliferation and differentiation of neural stem cells

Objective To observe the effects of conditioned medium from rat bone marrow mesenchymal stem cells (BMSCs)on the proliferation and differentiation of neural stem cells(NSCs)in vitro. Methods The NSCs were cultured with conditioned medium from BMSCs(BMSC-CM)under the condition of proliferation and differentiation respectively and shape variation of NSCs was observed. Then the proliferation of NSCs was analyzed by MTT,count-ing and diameter measurement of neurospheres;at the same time immunofluorescence and Western blot were used to identify their differentiation. Results A mass of neurospheres were obtained and cultured in suspension,those NSCs could differentiate into neurons,astrocytes and oligodendrocytes. The group of BMSC-CM promoted adhesion of neurospheres,but there was no obvious difference in OD value,the number and diameter of neurospheres com-pared with the control group. In addition,BMSC-CM could promote the expression of MBP(P < 0. 05),reduce the expression of GFAP(P < 0. 05)but had no effect on the expression of MAP-2. Conclusion BMSC-CM can pro-mote NSCs toward an oligodendrocyte fate but does not inhibit the proliferation of the NSCs.