从成年大鼠海马分离培养NG2蛋白聚糖阳性神经祖细胞的新方法

A new method for isolating NG2 proteoglycan expressing progenitor cells from the adult rat hippocampus

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 develop a simple method for isolating and culturing high purity NG2 proteoglycan expressing progenitor cells （NG2 cells） from adult rat hippocampus. Methods Hippocampus was dissected out from the adult female rats, digested with papain and centrifuged by Optiprep step density gradient. From the resulting major fraction, proliferating ceils were isolated and cultured in Neurobasal A containing B27 supplement and basic fibroblast growth factor 2 （ FGF2 ）, and then the characteristics of the cells were identified by double immunofluorescence stain. Results It was found that more than 90% of the culture were NG2 cells isolated from the adult rat hippocampus. These cells were neural stem like cells, and can be proliferated rapidly and passaged. Conclusion This study improved the primary culture method of adult NG2 cells in terms of purity and easiness.