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. 2023 Feb 2;10:102051. doi: 10.1016/j.mex.2023.102051

Fig. 5.

Fig 5

Isolation of PDGFRa+ OPCs enables the study of oligodendrocyte development. A. Transwell insert migration assay schematic and assessment of OPC migration in a control (no chemoattractant, intrinsic migration) versus chemoattractant (PDGF-AA, 20 ng/mL) driven migration. B. Proliferation assays that can be utilized to assess OPC proliferation include Ki67 staining and EdU uptake experiments. Once differentiated, oligodendrocytes in vitro can be used to assess myelination. C. Schematic of transplantation of OPCs into organotypic shiverer slice and immunofluorescence image demonstrating co-localization of MBP and calbindin, suggestive of axon ensheathment by transplanted OPCs, in shiverer slices collected at three weeks. D. Immunofluorescence image of positive MBP labeling in a nanofiber 3D cell-culture myelination assay, collected at two weeks. Antibodies: sheep anti-Ki67 (Novus, 1:200), rat anti- PDGFRα (BD, 1:100), rat anti-MBP (Novus, 1:500), rabbit anti-Calbindin (Swant, 1:5000), AlexaFluor 488 donkey anti-rat (ThermoFisher, 1:1000), AlexaFluor 594 donkey anti-sheep (Jackson ImmunoResearch, 1:500), AlexaFluor 647 donkey anti-rabbit (Jackson ImmunoResearch, 1:500).