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目的探索建立从成年大鼠海马分离培养高纯度NG2蛋白聚糖阳性神经祖细胞(NG2细胞)的简便方法。方法从成年雌性大鼠解剖出海马,经木瓜蛋白酶消化和Optiprep不连续梯度离心,从离心产生的组织细胞密集带,用含B27添加剂和碱性成纤维细胞生长因子2(FGF2)的NeurobasalA培养液,分离培养出增殖性细胞,免疫荧光双重染色法鉴定细胞性质。结果应用上述方法,可从成年大鼠海马简便地培养出纯度大于90%的NG2细胞,此细胞具有神经干细胞(NSCs)潜能,在FGF2作用下可迅速增殖并传代。结论此方法在纯度、产出率和简便性等方面改进了成体NG2细胞的原代培养技术。
Objective To establish a simple and convenient method for isolation and culture of high purity NG2 proteoglycan positive neural progenitor cells (NG2 cells) from the hippocampus of adult rats. Methods Hippocampus were dissected from adult female rats, and papillae were digested by papain and disrupted by Optiprep gradient centrifugation. Histoblastoid cells from centrifugation were stimulated with Neurobasal A containing B27 supplementation and basic fibroblast growth factor 2 (FGF2) , Proliferating cells were isolated and cultured, and the cell characteristics were identified by double immunofluorescence staining. Results Using the above method, NG2 cells with a purity of more than 90% can be easily cultured from the hippocampus of adult rats. The cells have the potential of neural stem cells (NSCs) and can proliferate rapidly and pass on under the action of FGF2. Conclusion This method improves the primary culture of adult NG2 cells in terms of purity, yield and convenience.