新生大鼠大脑O-2A细胞系的分离纯化和双向分化培养

Purification and Differentiation of Oligodendrocyte/Type-2 Astrocyte Lineage Cell from Neonatal Rat Cerebrum in Vitro

目的 在体外培养条件下获取纯度较高的O - 2A细胞系细胞并探索其分化规律。方法 根据星形胶质细胞、O - 2A祖细胞的生长时间差异及细胞的粘附特性不同 ,采用振荡法和差速贴壁法 ,结合 1型星形胶质细胞条件培养液 ,培养、分离纯化O - 2A祖细胞。结果 纯化的O - 2A祖细胞胞体呈椭圆形 ,具有典型的双极突起 ;经免疫细胞化学染色鉴定 ,细胞的纯度达 90 %。在有血清的情况下 ,O - 2A祖细胞定向分化为 2型星形胶质细胞 ;在无血清的情况下 ,定向分化为少突胶质细胞。结论 通过振荡法和差速贴壁法 ,结合 1型星形胶质细胞条件培养液分离纯化培养的O - 2A祖细胞具有双分化潜能。

Objective The present study is to obtain more pure oligodendrocyte /type-2 astrocyte progenitors (O-2A progenitors) and further understand the bipotential differentiation of O-2A progenitors in vitro. Methods Based on the different properties of cellular adhesions and developmental time-course, O-2A progenitors were isolated and purified with a standard shaking method and the different adhesion method combining with the type-1 astrocyte conditioned medium. Results The purified cells presented typical shape in which their bodies were oval and had two processes. Identified by immunohistochemistry, the purity of O-2A progenitors was 90%; and in serum-containing cultures these O-2A progenitors differentiated into type-2 astrocytes, whereas in serum-free condition they differentiated into oligodendrocytes. Conclusion A method can be established in which enough O-2A progenitors are required and O-2A progenitors have the capacity of bipotential differentiation.