新生小鼠视神经少突胶质细胞系定向分化培养及鉴定  

Purification and differentiation of oligodendrocyte lineage cell from the optic nerve in P_2 mice

作　　者：卜曙阳[1] 黄强 徐国旭[1] 周丽英 

机构地区：[1]苏州大学附属第二医院眼科,215004 [2]江苏省神经科学研究所

出　　处：《江苏医药》2005年第7期527-529,共3页Jiangsu Medical Journal

摘　　要：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O2A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。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.

关 键 词：定向分化 视神经 新生小鼠 鉴定 少突胶质细胞 O-2A祖细胞 细胞系 免疫细胞化学方法 流式细胞仪检测 细胞原代培养 抗神经节苷脂 血清甲状腺素 组织块法 分离纯化 前体细胞 星形细胞 细胞分化 定向培养 振荡法 培养基 管内段 

分 类 号：R392.12[医药卫生—免疫学] R774.6[医药卫生—基础医学]