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目的比较两种不同的分离培养方法对大鼠神经干细胞(NSCs)增殖的影响。方法分离新生24h内的SD大鼠脑组织细胞,采用无血清培养技术进行体外扩增培养,分别以机械吹打法和胰酶消化法两种方法进行扩增培养、传代。免疫细胞化学法对NSCs及其分化后的细胞进行Nestin、NSE、GFAP、CNP的鉴定。结果分离培养的细胞具有自我更新和增殖能力及Nestin表达阳性,诱导后可分化为神经元、星形胶质细胞及少突胶质细胞。无论第1代或第2代比较,胰酶消化组NSCs的增殖明显高于机械吹打组。结论实验中分离培养的细胞为具有自我更新和增殖能力的NSCs,可诱导分化为终末神经细胞。在两种分离培养方法中,以胰酶消化法培养可获得更多的NSCs。
Objective To compare the effects of two different isolation and culture methods on the proliferation of neural stem cells (NSCs) in rats. Methods SD rat brain cells were isolated and cultured in vitro for 24 hours. The cells were cultured and expanded in vitro by serum-free culture. The cells were cultured and passaged by mechanical blowing and trypsin digestion respectively. Immunocytochemistry was used to identify Nestin, NSE, GFAP and CNP in NSCs and their differentiated cells. Results The cells cultured in vitro showed the ability of self-renewal and proliferation, positive expression of Nestin and differentiation into neurons, astrocytes and oligodendrocytes. The proliferation of NSCs in trypsin digestion group was significantly higher than that in mechanical pipetting group, no matter in the first generation or the second generation. Conclusion The isolated and cultured cells in this experiment are self-renewing and proliferating NSCs that can differentiate into terminal neurons. In both isolation and culture methods, more NSCs can be obtained by trypsin digestion.