摘要
在体外培养条件下获取纯度较高的少突胶质细胞系细胞并探索其分化规律。新生大鼠大脑皮质原代混合胶质细胞培养 7～ 9d或 14～ 15d ,利用振荡和差速贴壁分离纯化少突胶质前体细胞 ,再进行无血清化学条件培养基培养。原代培养 7～ 9d时细胞分层形成 ,O 2A祖细胞 (oligodendrocyte/type 2astrocyteprogenitor)分散位于星形胶质细胞单层上 ,振荡分离后纯化率大于 95% ,分离后的O 2A祖细胞神经节苷脂GD3抗体阳性 ,无血清培养 3d后分化为“蜘蛛状”细胞 ,半乳糖脑苷脂抗体阳性。原代培养 14～ 15d时 ,星形胶质细胞层上出现葡萄样前O 2A祖细胞和少量星形胶质细胞。混合胶质细胞培养时细胞完全分层的出现是振荡分离O 2A祖细胞的适宜条件。分离纯化的O
The present study is to obtain more pure oligodendrocytes and further understand the committed differentiation of oligodendrocyte lineage cell in vitro. The primary mixed glial cells from neonatal rat cerebral tissue were cultured for 7～9 days or 14～15 days in vitro. The oligodendrocyte precursors were separated by orbital shaker, further purified by differential adhesion, and finally cultured in chemically serum free medium. The stratification of glial cells appeared and the O 2A progenitors (oligodendrocyte/type 2 astrocyte progenitor) scattered on the astrocyte monolayer in 7～9 days. The preparation of oligodendrocyte lineage cells were > 95%. The dissociated O 2A progenitors were identified by ganglioside GD3 antibody and further differentiated into so called “a spider web” oligodendrocyte marked by anti galactocerebroside in serum free medium for 3 days. The pre O 2A progenitors similar to a bunch of grapes can be seen and a few astrocytes came out on the astrocytes feeder layer in 14～15 days in vitro. The optimal procedure for cell isolation should be at the onset of complete stratification in the mixed glial cells culture. The O 2A progenitor can be spontaneously differentiated into oligodendrocyte.
基金
复旦大学医学院神经生物学国家重点实验室资助
中科院上海生理所开放实验室资助
