Advances in rodent oligodendrocyte precursor cells isolation and culture: From traditional methods to modern...

Oligodendrocyte progenitor cells (OPCs) are responsible for the generation of myelinating oligodendrocytes and are essential for myelination and repair of the central nervous system (CNS). In past decades, many methods have been presented for the reliable isolation and culture of OPCs to facilitate research into oligodendroglial biology and the development of effective therapeutic strategies such as cell transplantation and drug therapy in demyelinating and neurodegenerative diseases. However, challenges of purity, yield, and phenotypic stability still persist for each method.

A literature review was conducted using related keywords in the Scopus, Google Scholar, PubMed, and PubMed/Medline databases from the date of beginning until 1 November 2025. We examined preclinical research in rodent models that fit our search parameters, including in vitro and in vivo rat or mouse models, for both neonatal and adult CNS studies.

This review systematically describes the methodological progress of OPC isolation from mechanical isolation methods to advanced immunopanning culture systems, magnetic-activated cell sorting (MACS), and serum-free or growth factor-based culture systems. Additionally, it mentions important novel methods, like 3D or normoxic culture models, that have improved cell purity, function, and physiological relevance.

This article compares the principles, advantages, and disadvantages of each method to create a single set of guidelines for selecting the best methods regarding the equipment and goals of an investigation. It also suggests ways to design OPC isolation systems that are more standardized, scalable, and ethically acceptable, which may accelerate in the use of these systems in regenerative neuroscience.