大鼠大脑皮层神经干细胞培养方法比较

发布时间：2018-03-01 03:28

本文关键词： SD大鼠大脑皮层神经干细胞培养方法　出处：《神经解剖学杂志》2013年01期 　论文类型：期刊论文

【摘要】：目的:比较不同培养基对大脑皮层神经干细胞增殖的影响,以寻找培养神经干细胞的有效方法。方法:体外分离生后24 h SD大鼠大脑皮层细胞,使用DMEM/F12+B27+bFGF、Neurobasal+B27+bFGF和Neuro-basal+B27三种培养条件,分别用悬浮培养法和贴壁培养法对细胞进行培养。免疫细胞化学技术分别以Nestin、β-TubIII和GFAP标记神经干细胞、神经元和星形胶质细胞。结果:在悬浮培养情况下,三种培养条件均诱导所培养的细胞形成神经球,经染色鉴定神经球中的细胞均表达Nestin;其中DMEM/F12+B27+bFGF培养的神经干细胞增殖速度最快(P0.01),Neurobasal+B27培养的神经干细胞增殖效率最高(P0.01)。在贴壁培养情况下,经染色鉴定,部分细胞表达β-TubⅢ或GFAP。结论:大脑皮层神经干细胞不适合贴壁培养,DMEM/F12+B27+bFGF条件适合大脑皮层神经干细胞体外增殖。
[Abstract]:Objective: to compare the effects of different culture media on the proliferation of neural stem cells (NSCs) in cerebral cortex, and to find an effective way to culture NSCs. Methods: the cortical cells of SD rats were isolated 24 hours after birth in vitro. DMEM/F12 B27 bFGFG Neurobasal B27 bFGF and Neuro-basal B27 were used to culture the cells by suspension culture and adherent culture respectively. Nestin, 尾 -TubIII and GFAP were used to label Nestin, 尾 -TubIII and GFAP by immunocytochemistry, respectively. Results: under the condition of suspension culture, the cultured cells were induced to form neurospheres. Nestin was detected by staining, and the proliferation rate of neural stem cells cultured with DMEM/F12 B27 bFGF was the fastest. The proliferation efficiency of neural stem cells cultured in neurobasal B27 was the highest (P 0.01). Under the condition of adherent culture, the proliferation rate of neural stem cells was identified by staining. Conclusion: the neural stem cells of cerebral cortex are not suitable for the proliferation of neural stem cells in vitro under the condition of DMEM / F12B27 bFGF.
【作者单位】：上海体育学院运动科学院;
【基金】：国家自然科学基金(30870349) 运动健身科技省部共建教育部重点实验室基金(上海体育学院)
【分类号】：R329

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