【摘要】 目的　体外诱导大鼠骨髓间质干细胞 (rMSC)分化为神经细胞。方法　用SD大鼠股骨骨髓细胞体外培养。用绞股蓝总甙加入无血清L DMEM诱导MSC分化为神经细胞。免疫细胞化学鉴定有神经元烯醇化酶 (NSE)、神经干细胞标志物巢蛋白 (nestin)和胶质纤维酸性蛋白 (GFAP)的表达。结果　大鼠骨髓间质干细胞在体外扩增 5～ 2 2代 ,对照组不加任何诱导剂 ,MSC可分化为神经样细胞 (5 3 1%± 4 3% )。加入绞股蓝诱导剂诱导 1～ 5h ,MSC形态转变为典型的神经样细胞(90 0 %± 4 6 % )。免疫细胞化学染色显示诱导出的神经样细胞 ,NSE、nestin、GFAP表达阳性。继续培养 12～ 19d ,5d后对照组所有细胞逐渐死亡。绞股蓝组持续诱导 12～ 19d后神经样细胞形态完好 ,大部分细胞变为圆形并聚集成团 ,NSE、nestin、GFAP表达阳性。如果换回正常培养液 ,神经样细胞可逆转回MSC并传代。结论　骨髓本身可能存在神经干细胞。大鼠骨髓间质干细胞用中成药绞股蓝诱导可分化为多种形态的神经样细胞。更多还原

【Abstract】 Objective To investigate the differentiation of rat mesenehymal stem cells (rMSC) into neuron-like cells. Methods rMSC were separated from femur marrow and expanded in L-DMEM culture medium supplemented with 10% FSC. rMSC were induced to differentiate into neurons by L-DMEM/gypenosides. At the same time the rMSC were cultured in L-DMEM in control group. Nestin, neuron-specific enclose (NSE), and glial fibrillary acidic protein (GFAP) were detected by immunocytochemistry method. Results rMSC were expanded as undifferentiated cells in culture from 5 to 22 passages, indicating their differentiated capacity. The simple method had induced the rMSC to exhibit a neuronal phenotype, expressing a positive NSE, nestin, and GFAP, at 3 hours. The differentiating cells (treated group 90.0%±4.6%), and undifferentiating cells (control group 53.1%±4.3%), had expressed characteristics of various neuronal cells at 3 hours, but the trait was undetectable at 5 days in control group. In contrast, the expression of nestin, NSE, GFAP in gypenosides group, persisted on from 3 hours to 19 days, and expressed the characteristics of neuronal precursor stem cells. Conclusion It suggests that rat neural stem cells (rNSC) might exist in bone marrow. rMSC should be differentiated into neuronal precursor stem cells as induced by Chinese medicine-gypenosides in vitro.更多还原