Culture and isolation of oligodendrocyte precursor cells from neonatal rat cerebral cortices / 中国矫形外科杂志

ABSTRACT

[Objective]To observe the growth pattern of oligodendrocyte precursor cells(OPCs) in the primary culture from neonatal rat cerebral cortices and study the methods of cell culture and isolation in order to obtain purified OPCs for the experiments of cell transplantation.[Method]The mixed glial cells from the cerebral cortices of 48-hour-old Sprague-Dawley(SD) rats were cultured in vitro.Until 9-10 days in vitro OPCs were isolated and purified by the shaking process and differential adhesion,and then continued OPCs culture in the defined medium.The growth pattern of OPCs in vitro was investigated by contrast phase microscopy and OPC s were further identified with the immunocytochemical techniques.[Result]The distinct stratification of OPCs and astrocytes developed around 9-10 days in primary culture.At this point,the OPCs scattered on the top of the monolayer astroctyes and the soma of most OPCs typically appeared oval or round with two or three processes.The purity of the isolated OPCs reached 95% and these OPCs further developed into the mature oligdendrocytes which were immunoreactive to the specific antigen of oligodendrocyte lineage cells,Oligo2.[Conclusion]OPCs separated from the cerebral cortices of neonatal SD rats can be maintained as immature precursor cells in culture,and OPCs are able to be purified by shaking and differential adhesion under the condition of appropriate cell stratification.