Objective: To explore the methods of isolation and purification of oligodendrocyte precursor cells (OPCs) in the brain of newborn Sprague-Dawley(SD) rats, and to explore the effects of different intervention times of oxygen glucose deprivation(OGD) on oligodendrocyte viability. Methods: The high purity oligodendrocyte precursor cells were obtained from the cerebral cortex of SD neonatal rats by differential adhesion, oscillatory separation and purification, and restricted medium directional culture method. The morphological changes of oligodendrocyte precursor cells during differentiation were observed under microscope. The purified oligodendrocyte precursor cells, mature oligodendrocytes, astrocytes and microglia were labeled with specific antibodies A2B5, MBP, GFAP and IBA _1 to analyze the purity of the purified cells. The effect of OGD on the proliferation of oligodendrocyte cell line was observed by CCK-8 assay. Results: Specific antibodies A2B5, MBP, GFAP and IGB-1 were used to label oligodendrocytes, mature oligodendrocytes, astrocytes and microglia. A2B5, MBP positive cells accounted for 96.2%, 2.1%, GFAP and IBA-1 staining positive cells were less than 3%. The results of MTT detection showed that in OGD model, the cell viability decreased with the prolongation of OGD intervention time. Conclusion: (1) High purity oligodendrocyte precursor cells can be obtained by combination of differential adhesion, constant temperature oscillation separation and conditioned medium. (2) The OGD model established in vitro can cause the damage of oligodendrocyte precursor cells and decrease the activity of oligodendrocyte precursor cells. 3 h intervention is suitable for the establishment of OGD model of oligodendrocyte precursor cells in vitro. |
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