抗氧化剂在分离培养成年大鼠海马神经祖细胞中的作用

The effect of antioxidants on the isolation and culture of adult rat hippocampal neural progenitor cells

目的探讨抗氧化剂在分离培养成年大鼠海马NG2蛋白聚糖阳性神经祖细胞(NG2细胞)中的重要作用。方法根据文献从成年大鼠海马分离培养NG2细胞的方法,以B27-AO添加剂(去除B27添加剂中所有抗氧化剂的衍生物)替代原培养液中的B27添加剂,并将四种抗氧化剂单独或组合加入B27-AO培养环境,行海马细胞原代及传代培养。以乳酸脱氢酶(LDH)分析法测定细胞活性;以免疫荧光双重染色法鉴定细胞性质。结果B27-AO添加剂未能分离或传代大部分NG2细胞,而过氧化氢酶(Catalase)能挽救大部分在B27-AO培养条件下死亡的细胞。结论抗氧化剂在NG2细胞分离培养过程中是必不可少的。

Objective To explore the effect of antioxidants on the isolation and culture of adult rat hippocampal NG2 proteoglycan expressing progenitor cells （NG2 cells）. Methods The primary culture was done according to the reference for isolation of NG2 cells from the adult rat hippocampus. Instead of B27 supplement, B27-AO supplement （a derivative of B27 supplement that lacks all the antioxidants） was used, and four types of antioxidants were added to B27-AO culture conditons separately or combined. Cell activities were determined by Lactate dehydrogenase （LDH） assay, and the characteristics of the cells were identified by double immunofluorescence stain. Results B27-AO supplement failed to expand or passage most of the NG2 cells, and catalase rescued most of the cells in B27-AO culture condition. Conclusion Antioxidants are essential for the isolation and culture of NG2 cells.