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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 investigate the important role of antioxidants in the isolation and culture of NG2 proteoglycan-positive neural progenitor cells (NG2 cells) from hippocampus of adult rats. Methods According to the method of isolating and culturing NG2 cells from the adult rat hippocampus, the B27 additive in the original culture solution was replaced by the B27-AO additive (a derivative of all the antioxidants in the B27 additive), and the four antioxidants alone or in combination Join B27-AO culture environment, primary and hippocampal cell subculture. Cell viability was determined by lactate dehydrogenase (LDH) assay; cell characteristics were identified by double immunofluorescence staining. Results The B27-AO additive failed to isolate or pass most of the NG2 cells, whereas catalase blocked most of the cells that died under the B27-AO culture conditions. Conclusion Antioxidants are essential for NG2 cell isolation and culture.