Isolation and purification of mouse OPCs was performed according to previously described immunopanning protocols using an anti-Pdgfra (CD140a) antibody for positive selection of OPCs (Shiow et al., 2017 (link); Yuen et al., 2014 (link); Dugas and Emery, 2013 (link)). Briefly, OPCs were immunopanned from P7-P9 mouse cortices and plated on poly-D-lysine coverslips (Neuvitro). Cells were kept in proliferation media (PDGF-AA, CNTF, and NT3; Peprotech) at 10% CO2 and 37°C. After two days in proliferation media, differentiation was induced by changing media to contain CNTF and triiodothyronine (T3; Sigma). Note that mycoplasma contamination testing was negative.
Other organizations :
Medical Research Council, University of California, San Francisco, University of Cambridge, Wellcome/MRC Cambridge Stem Cell Institute, Max Planck Institute of Experimental Medicine, Nanoscale Microscopy and Molecular Physiology of the Brain Cluster of Excellence 171 — DFG Research Center 103
Change in media composition (from proliferation media to CNTF and T3)
dependent variables
OPC differentiation
control variables
Cell type (OPCs from P7-P9 mouse cortices)
Cell culture conditions (10% CO2 and 37°C)
Coating surface (poly-D-lysine coverslips)
Mycoplasma contamination (tested negative)
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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