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.