新生小鼠视神经少突胶质细胞系定向分化培养及鉴定

目的 探索P2期小鼠视神经少突胶质细胞的分化规律。方法 经颅开眶法获取P2期小鼠管内段视神经2mm，组织块法培养，振荡法分离纯化，化学限定性培养基定向分化培养纯化，免疫细胞化学方法鉴定，流式细胞仪检测纯度。结果 混合胶质细胞原代培养7d时细胞分层，少突胶质细胞前体细胞(O-2A)分散于星形细胞层上，分离后的O-2A祖细胞抗神经节苷脂GD3阳性；无血清甲状腺素限定性培养基定向培养2周后O-2A祖细胞分化为MBP阳性成熟的少突胶质细胞细胞。结论 组织块法培养、振荡法分离纯化P2期小鼠视神经少突胶质细胞前体细胞，纯度高。O-2A祖细胞可分化为成熟少突胶质细胞。

Purification and differentiation of oligodendrocyte lineage cell from the optic nerve in P2 mice

Objective To obtain the purified oligodendrocyte progenitors from the optic nerve in optic canal of P_2 mice and study the committed differentiation of the oligodendrocyte lineage cells in vitro.Methods A segment of 2 mm optic nerve in optic canal was obtained from the mice in P_2 period via craiotomy.The oligodendrocytes were separated by orbital shaker,further purified by differential adhension and finally cultured in chemically serum-free medium with throid. O-2A progenitors and oligodendrocytes were identified by microscopic examination, immunohistochemisty through ABC assay and flow cytometry.Results The stratification of the primary mixed glial cells from the optic nerves appeared and the O-2A progenitors scattered on the astrocyte monolayer on the 7th day.The O-2A progenitors were identified by ganglioside GD_3 antibody .Oligodendrocytes showed positive myelin basic protein (MBP). Conclusion Shaking the cultures and CDM culture are the easy,quick and reliable approach to separate and purify the astrocytes and oligodendrocytes progenitor from primary mixed glial cells of the optic nerve in P_2 mice.The 0-2A progenitor can spontaneously differentiate into oligodendrocyte.