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Isolation and culture of rat and mouse oligodendrocyte precursor cells.

Nature protocols (2007-06-05)
Ying Chen, Veerakumar Balasubramaniyan, Jie Peng, Edward C Hurlock, Michelle Tallquist, Jianrong Li, Q Richard Lu
ABSTRACT

The ability to isolate oligodendroglial precursor cells (OPCs) provides a powerful means to characterize their differentiation, properties and potential for myelin repair. Although much knowledge is available for isolation of OPCs from the rat central nervous system, preparation and maintenance of mouse OPCs has been until recently a challenge owing to difficulties in obtaining a sufficient quantity of purified OPCs. Here, we describe protocols to prepare highly enriched rat OPCs and nearly homogenous mouse OPCs. The mouse method generates predominantly OPCs from cortical neural progenitor cells as clonal aggregates called "oligospheres" by taking advantage of molecular genetic tools. Isolated OPCs can be further differentiated into oligodendrocytes. Collectively, we describe simple and efficient methods for the preparation and in vitro maintenance of enriched OPCs from rats and mice. Isolation and culture of a large, homogenous population of rodent OPCs should significantly facilitate studies on OPC lineage progression and their utility in myelin repair after injury.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Biotin, ≥99% (HPLC), lyophilized powder
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
N-Acetyl-L-cysteine, BioXtra, ≥99% (TLC)