大鼠脊髓星形胶质细胞的分离与纯化

Purification and identification of astrocytes from rats spinal cord

目的:纯化培养和鉴定新生大鼠的脊髓星形胶质细胞,建立稳定的星形胶质细胞培养体系,为研究创伤后慢性神经病理性痛的脊髓机制奠定基础。方法:根据成纤维细胞与星形胶质细胞生长倍增时间的差异,用阿糖胞苷(Ara-C)抑制成纤维细胞生长分裂的方法获取星形胶质细胞,用胶质原纤维酸性蛋白(GFAP)免疫荧光法对其鉴定。同时与差速贴壁培养法进行对比。结果:用Ara-C处理后,可获得较高纯度的星形胶质细胞,经鉴定其纯度可达90%以上,而对照组几乎全部为成纤维细胞。结论:Ara-C处理能彻底清除成纤维细胞,从而保持星形胶质细胞的正常生长繁殖,达到纯化星形胶质细胞的目的。

Objiective： To find out the method for purifing astrocytes from the spinal cord of neonatal SD rats, and establish a pure cell culture system for the investigation of the effect of the spinal cord on chronic neuropathic pain development. Methods： According to the different population doubling time between astrocyte and fibroblast, cells were cultured with suppression of fibroblasts （Ara-c） and identified with immumofluorescence method against glial fibrillary acidic protein （GFAP）. The adherent culture with different time was used as the control group. Results： In the Ara-C treated group, a pure culture of astrocyte cells was obtained. After being identified, the purity was over 90%. In contrast, there were almost full of fibroblasts in the control group. Conclusion： Ara-C can suppresses the proliferation of fibroblasts, and enable to maintain the normal growth of astrocyte cells.