视神经胶质细胞的培养及其特征观察

Culture of glial cells of optic nerve in mice

目的 体外培养、分离、纯化P2期小鼠视神经胶质细胞,并观察其生物学特性。方法 纤颅开眶获取P2期小鼠管内段视神经2mm,组织块法培养,振荡法分离纯化,免疫组织化学方法鉴定,流式细胞仪检测纯度。结果 视神经星形胶质细胞体外迅速分裂增殖,少突胶质细胞生长缓慢,传代困难,振荡法分离纯化的细胞纯度高,星形胶质细胞GFAP染色阳性,阳性率96.2％,少突胶质细胞MBP染色阳性,阳性率96％。结论 组织块法培养、振荡法分离纯化P2期小鼠视神经星形和少突胶质细胞纯度高,是进一步研究视神经胶质细胞的结构和功能的叶靠材料。

Objective To obtain the purified astrocytes and oligodendrocyles from the optic nerve of optic canal of mice in P2 period and observe their features. Methods Optic nerve in optic canal of mice in P2 period was obtained via craiotorny. Mixed glial cells were primary cultured. Cultures were shaken to allow detachment of certain cells. The astrocyles and oligodendroeytes were identified by microscopic examination and immunohistochemistry through ABC assay and flow cytometric. The results were analyzed. Results Astrocytes proliferated rapidly. Preparations appeared to be (96.2±2.8)% pure. The astrocytes were showed positive reaction to glial fibnlllary acidic protein ( GFAP). Oligodendrocyles showed difficult in proliferating. The oligodendrocytes were showed postive to rnyelin basic protein (MBP). Conclusion Shaking the cultures is an easy, quick and reliable approach to separation and purification of astrocytes and oligodendrocytes from primary mixed glial cells of optic nerve in P, periods mices. This preparations should significantly aid in effort to examine the biochemistry, physiology, pharmacology of this class of optic nevers particularly aid in the efforts to study the role of astrocytes after injury of optic never.