Isolation and Culture of Oligodendrocyte Lineage Cells from Rodent Brains

Oligodendrocytes (OLs) are central nervous system (CNS) glia that serve the irreplaceable function of axon myelination. In vitro primary OL cultures have greatly enhanced our insight into OL physiology, such as the role of certain cytokines and growth factors in OL progenitor cell (OPC) proliferation and OL differentiation. Methods widely used for the isolation of primary OPCs from neonatal rodent brains, such as the "shaking" method or immunopanning, have been in use for an extended period of time without meaningful updates, and while they still offer a robust, high-quality OL culture, low cost-effectiveness and high technical demand make such methods unsuitable for mass production. In this segment, we present a simple and cost-effective method to isolate OPCs from neonatal rodent brains by a two-step differential centrifugation process, followed by the use of optimized media formulations to boost OPC proliferation and achieve a high cell yield. Additionally, we demonstrate a method to cryopreserve OPCs cultivated through this method, which may assist in taking full advantage of the high cell numbers that this method provides.