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A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

机译:大鼠背根神经节来源高纯度卫星神经胶质细胞的原代培养新方法

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摘要

Satellite glial cells surround neurons within dorsal root ganglia. Previous studies have focused on single-cell suspensions of cultured neurons derived from rat dorsal root ganglia. At present, the primary culture method for satellite glial cells derived from rat dorsal root ganglia requires no digestion skill. Hence, the aim of the present study was to establish a novel primary culture method for satellite glial cells derived from dorsal root ganglia. Neonatal rat spine was collected and an incision made to expose the transverse protrusion and remove dorsal root ganglia. Dorsal root ganglia were freed from nerve fibers, connective tissue, and capsule membranes, then rinsed and transferred to 6-well plates, and cultured in a humidified 5% COsub2/sub incubator at 37°C. After 3 days in culture, some cells had migrated from dorsal root ganglia. After subculture, cells were identified by immunofluorescence labeling for three satellite glial cell-specific markers: glutamine synthetase, glial fibrillary acidic protein, and S100β. Cultured cells expressed glutamine synthetase, glial fibrillary acidic protein, and S100β, suggesting they are satellite glial cells with a purity of 95%. Thus, we have successfully established a novel primary culture method for obtaining high-purity satellite glial cells from rat dorsal root ganglia without digestion.
机译:附属神经胶质细胞围绕背根神经节内的神经元。先前的研究集中在源自大鼠背根神经节的培养神经元的单细胞悬液。目前,源自大鼠背根神经节的卫星神经胶质细胞的原代培养方法不需要任何消化技能。因此,本研究的目的是为源自背根神经节的卫星神经胶质细胞建立一种新颖的原代培养方法。收集新生大鼠的脊柱并切开切口以暴露横向突起并去除背根神经节。清除背根神经节的神经纤维,结缔组织和囊膜,然后冲洗并转移到6孔板中,并在37°C的5%CO 2 湿箱中培养。培养3天后,一些细胞已经从背根神经节迁移。传代培养后,通过免疫荧光标记的三种卫星神经胶质细胞特异性标记物鉴定细胞:谷氨酰胺合成酶,神经胶质纤维酸性蛋白和S100β。培养的细胞表达谷氨酰胺合成酶,神经胶质原纤维酸性蛋白和S100β,表明它们是卫星神经胶质细胞,纯度> 95%。因此,我们成功地建立了一种新的原代培养方法,该方法无需消化即可从大鼠背根神经节获得高纯度的卫星神经胶质细胞。

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