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目的探索从成年大鼠中枢神经系统(CNS)不同区域是否能分离培养出NG2蛋白聚糖阳性神经祖细胞(NG2细胞)。方法从成年雌性大鼠解剖出CNS的9个不同区域,分别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,再以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CNS的9个不同区域分离出具神经干细胞(NSCs)潜能的NG2细胞。结论成年大鼠CNS的非神经发生区域同样存在NSCs样细胞,且可通过适当方法体外培养。
Objective To explore whether NG2 proteoglycan positive neural progenitor cells (NG2 cells) can be isolated and cultured from different regions of central nervous system (CNS) of adult rats. Methods Nine different regions of the CNS were dissected from adult female rats and were isolated by papain digestion and Optiprep discontinuous gradient centrifugation, respectively. The tissue-cell-rich bands from centrifugation were stained with B27-supplemented and basic fibroblast growth factor 2 ( FGF2) NeurobasalA broth, proliferating cells were isolated and cultured, and then identified by immunofluorescence double staining of cell properties. Results Using the above method, NG2 cells capable of developing neural stem cells (NSCs) were isolated from 9 different regions of adult rat CNS. Conclusion NSCs-like cells also exist in the non-neurogenic region of adult rat CNS and can be cultured in vitro by appropriate methods.