Isolation of neural stem and oligodendrocyte progenitor cells from the brain of live rats.

Stem Cell Reports

Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, CB2 0AW Cambridge, UK; Lab of Developmental Biology, Department of Biology, University of Patras, 26504 Patras, Greece. Electronic address:

Published: October 2021

Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term "milking." The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8514974PMC
http://dx.doi.org/10.1016/j.stemcr.2021.08.015DOI Listing

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