Improvement in isolation and culture method for oligodendrocyte precursor cells and establishment of the in vitro model for lineage restricted oligodendrocyte differentiation

Abstract

To establish the improved methods for oligodendrocyte precursor cells (OPCs) isolation, culture and subculture and to investigate the characteristics of OPCs differentiation and proliferation in vitro. Methods: OPCs isolated from neonatal SD rat spinal cords were cultured in vitro. Cell proliferation was induced by the culture solution with basic fibroblast growth factor and platelet-derived growth factor, and the proliferation of OPCs was determined by MTT assay. OPCs differentiation to mature oligodendrocytes was induced by triiodothyronine and ciliary neurotrophic factor. The morphology and characteristics of OPCs and differentiated cells were observed via microscope and immunostaining. Results: More than 95% of the cells exhibited typical OPCs morphology with bipolar or tripolar bumps and expressed A2B5, while the OPCs kept the proliferative capacity in vitro. After culture for differentiation, the OPCs progressively differentiated into mature oligodendrocytes which specifically expressed myelin basic protein. Conclusion: The OPCs retain the ability of differentiation and proliferation in vitro.