目的 为在体外考察微管结合蛋白STOP对孤独症谱系障碍BTBR小鼠少突胶质细胞髓鞘形成的影响，建立一种较高纯化率的BTBR小鼠大脑皮质少突胶质前体细胞的原代培养方法，利用慢病毒载体建立一种高转染效率的BTBR小鼠大脑皮质少突胶质前体细胞过表达STOP基因的转染方法。  方法 以BTBR乳鼠作为实验对象，每次取6~10只，独立重复3次，0.25%胰蛋白酶消化制备成单细胞悬液，采用免疫磁珠细胞分选法获得原代少突胶质前体细胞。取原代培养的少突胶质前体细胞，利用我们前期构建的STOP基因载体进行转染实验，转染72~96 h后，在荧光显微镜下观察细胞携带荧光的显色情况。  结果 采用免疫磁珠细胞分选法提取的BTBR小鼠大脑皮质原代少突胶前体细胞在48 h后基本完全贴壁，细胞生长状态好，培养第5天，免疫荧光法鉴定显示，细胞纯度极高可达95%。建立了一种转染效率较高的BTBR小鼠原代少突胶质前体细胞的慢病毒转染方法，高内涵显微镜拍摄后显示，细胞荧光表达明显，将少突胶质前体细胞的慢病毒转染率提高到60%~70%。  结论 成功分选培养获得纯度较高的BTBR小鼠大脑皮质原代少突胶质前体细胞，成功建立了一种转染率较高的慢病毒感染BTBR小鼠大脑皮质原代少突胶质前体细胞的方法。

Objective To investigate the effect of microtubule binding protein STOP on myelin formation of oligodendrocyte in BTBR mice spectrum disorder in vitro, a highly purified primary culture method  of oligodendrocyte precursor cells from cerebral cortex of BTBR mice was established. Establishment of a highly efficient transfection method  for overexpression of STOP gene in oligodendrocyte precursor cells of BTBR mice cerebral cortex using lentiviral vector.   Methods BTBR mice were used as experimental objects, 6-10 suckling mice were taken each time, repeat 3 times independently. The single cell suspension was prepared by trypsin digestion, and the primary oligodendrocyte precursor cells were obtained by immunomagnetic bead cell sorting method . After 5 days of culture, the cell purity was identified by oligodendrocyte precursor cell marker staining. The primary cultured oligodendrocyte precursor cells were transfected with STOP gene vector constructed in the early stage of the project group. 72-96 hours after transfection, the fluorescence staining of oligodendrocyte precursor cells was observed under fluorescence microscope, and the transfection rate and cell survival rate were calculated.   Results The oligodendrocyte precursor cells of BTBR mice extracted by immunomagnetic beads sorting method  basically adhered to the wall completely after 48 hours, and the cells had strong ability of proliferation. On the fifth day of culture, the purity of the cells was more than 95% identified by immunofluorescence. A lentivirus transfection method  for primary oligodendrocyte precursor cells of BTBR mice with high transfection efficiency was established. The fluorescence expression of the cells was obvious after being photographed by high connotation microscope, the lentivirus transfection rate of oligodendrocyte precursor cells was increased to 60%-70%.   Conclusion The primary oligodendrocyte precursor cells of BTBR mouse cerebral cortex with high purity were successfully isolated and cultured. A method  for lentivirus infection of primary oligodendrocyte precursor cells in the cerebral cortex of BTBR mice is successfully established.