A long-term and economical method for culturing human oligodendrocyte precursor cells

Abstract

Objective Cell therapy is a possible effective way to treat myelination disorder diseases.Cell therapy needs to apply a method for culturing oligodendrocyte precursor cells.Therefore, this study was to develop a stable, efficient and economical method for obtaining human oligodendrocyte precursor cells(OPCs) in order to provide cell required for clinical treatment.And this will provide a new option for clinical applications. Methods Human OPCs were obtained through magnetic bead sorting and cultured in OPCs proliferation medium.For a short-time(2, 4, 6, 8 days) and long-time culture, morphology of OPCs was observed.The fourth generation of OPCs was analyzed for expression of OPCs specific markers O4, Sox10 and platelet derived growth factor alpla receptors(PDGFαR) and the capacity to differentiate into oligodendrocytes by immunofluorescence staining.At the same time, the effects of B27 and N1 on OPCs growth state were inspected as well. Results For a short-time culture, OPCs had typical bipolar or tripolar morphology and proliferated in good condition.For a long-time culture, all 4 generations OPCs had typical bipolar or tripolar morphology; the fourth generation OPCs highly(>90%) expressed O4, Sox10 and PDGFαR, after induction, OPCs could be differentiated into oligodendrocytes.After 4 generations of long-time culture, OPCs already maintained the ori-ginal sharp, high purity and had the capacity to differentiate into oligodendrocytes.It was indicated that this culture system was suitable for human OPCs for a long-time culture. Conclusions Overall, using this culture system, isolated human OPCs not only can be stably cultured and proliferated in vitro, but also have the capacity to differentiate into oligodendrocytes.From this reproducible method, a large number of human OPCs can be stably obtained in vitro as conve-nient as possible.And this will provide a new option for clinical applications.This method uses fewer cytokines.Therefore, this method will provide stable, efficient and economical OPCs for cell therapy of myelination disorders or myelin damage diseases. Key words: White matter damage; Oligodendrocyte precursor cells; Proliferation; Oligodendrocyte