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为建立有效纯度高的恒河猴星形胶质细胞体外培养体系,无菌条件下取6月龄恒河猴婴猴的大脑皮质,除去白质,充分剪碎后机械吹打制成细胞悬液接种培养,待原代培养的细胞长满培养皿后,通过恒温摇床振荡和传代差速贴壁除去寡突胶质细胞、成纤维细胞等,得到纯化后的星形胶质细胞,并用GFAP-FITC(glial fibrillary acidic protein-fluorescein isothiocyanate)免疫荧光法对所获细胞进行鉴定。分离培养的细胞具备典型的星形胶质细胞形态,并表达星形胶质细胞特异性抗原GFAP(glial fibrillary acidic protein),纯化后获得了高纯度的恒河猴星形胶质细胞。采用该培养方法成功建立原代恒河猴星形胶质细胞培养体系,为体外研究嗜神经性病毒、神经系统疾病及其相关中枢神经病理机制等提供了可靠的细胞模型。
In order to establish an effective culture system of rhesus monkey astrocytes with high purity and purity, the cerebral cortex of 6-month-old rhesus monkey was removed under sterile conditions to remove white matter, After the primary cultured cells are covered with petri dishes, oligodendrocytes, fibroblasts and the like are eliminated by oscillating at a constant temperature and differentiating and passaged at a differential transmission rate to obtain purified astrocytes, and cultured with GFAP- The cells were identified by FITC (glial fibrillary acidic protein-fluorescein isothiocyanate) immunofluorescence. The cultured cells have typical astrocyte morphology and expressed GFAP (glial fibrillary acidic protein), and obtained high purity rhesus monkey astrocytes after purification. The culture method was used to establish the primary culture system of rhesus monkey as a reliable cell model for studying the neurotropic virus, the nervous system diseases and the related central nervous pathological mechanism in vitro.