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(P<0.01),Neurobasal+B27培养的神经干细胞增殖效率最高(P<0.01)。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。
Objective： To look for an effective method for culturing neural stem cells（NSCs） of cerebral cortex. Meth- ods：In vitro, newborn 24 h SD rat cerebral cortex cells NSCs were gained. In the condition of DMEM/F12 ＋ B27 ＋ bF- GF, Neurobasal ＋ B27 ＋ bFGF and Neurobasal ＋ B27 to culture cells and use suspension culture method and adherent cul- ture method, respectively. Immunoeytochemical stainings of Nestin, β-TublII and GFAP was used to identify NSCs, neu- rons and astrocytes. Results： In the case of suspension, all three culturing conditions induced cells into neurospheres, and all of the cells expressed Nestin. While, DMEM/F12 ＋ B27 ＋ bFGF promoted proliferation speed of NSCs （ P 〈 0.01 ）, Neurobasal ＋ B27 promoted better proliferation efficiency （P 〈0.01 ）. In case of adherent culture, some cells ex- pressed 13-TublII or GFAP. Conclusion： Cerebral cortical NSCs are not suitable for adherent culture. Suspension culture with DMEM/F12 ＋ B27 ＋ bFGF is an appropriate condition to induce proliferation of cerebral cortical NSCs.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2013年第1期70-74,共5页
Chinese Journal of Neuroanatomy
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)
