An efficient and economical culture approach for the enrichment of purified oligodendrocyte progenitor cells

Oligodendrocyte progenitor cell (OPC) culture has provided a powerful approach to mechanistically investigate the proliferation and differentiation of oligodendroglia. However, existing culture methods (including the traditional shake-off method) have limitations, particularly their low productivities. Therefore, we developed a simplified and highly efficient method to produce a large yield of OPCs with low expense by using specialised modified media, in which B104-conditioned medium (B104-CM) instead of growth factors was used as a mitogenic source for OPC propagation, while a modified OPC isolation-medium was applied to improve the isolation of OPCs. First, we withdrew foetal bovine serum when primary mixed glial cultures were 65–75% confluent and substituted with modified oligodendrocyte growth medium to enrich OPCs. Second, we employed a chemical-based method to isolate and purify OPCs from mixed glial cultures using a modified oligodendrocyte isolation medium. As a result, our approach produced a high yield of purified OPCs, approximately 90-fold higher than that produced via the traditional shake-off method. Importantly, the purified OPCs produced via our modified approach maintained typical capacities of proliferation and differentiation observed in oligodendrocyte lineage cells. Together, our modified method provides a highly efficient approach to OPC culture for oligodendrocyte research.

► A simplified and highly efficient method to produce a large yield of purified OPCs at a low expense was developed.
► B104-conditioned medium was used as a mitogenic source for OPC propagation combined with an adopted paradigm of OPC culture.
► Two modified media were used to enrich for OPCs at an early stage of the mixed glial cultures and to promote cell detachment in isolation.
► A two-step chemical-based isolation procedure was used to increase the yield of purified OPCs.