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. 2018 Feb 1;8:2113. doi: 10.1038/s41598-018-19934-6

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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WmOPCs proliferate less and differentiate faster than gmOPCs. Oligodendrocyte progenitor cells (OPCs) isolated from the cortex (gmOPCs) and non-cortex (wmOPCs) of neonatal rat forebrains were cultured for 4 hours (a,b) or 48 hours in the presence of PDGF-AA and FGF-2 (c–i), followed by differentiation for 3 (immature stage, e,f and h) or 6 days (mature stage, g–i). (a,b) OPC migration towards a 10 ng/ml PDGF-AA gradient (4 hours) was determined using a transwell assay. Representative images of migrated DAPI-stained OPCs are shown in a; quantitative analyses of the percentage of DAPI-stained migrated OPCs of the total number of plated cells in (b) (n = 10), (c,d) OPC proliferation was determined by immunocytochemistry for the proliferation marker ki67. Representative images are shown in (c), quantitative analyses of the number of ki67-positive of total DAPI-stained cells in (d) (n = 16, at least 150 cells analysed per independent experiment). Note the higher percentage of proliferating gmOPCs compared to wmOPCs. (e–i) OPC were differentiated for 3 (e,f, and h) and 6 days (g–i) and incubated with either (e) R-mAb, recognizing GalCer/sulfatide, or (f–i) double stained for MBP (red), a mature marker of oligodendrocytes (OLGs) and Olig2 (green), OLG lineage marker. Representative images are shown in (e,f) and (g); quantitative analyses of the number of MBP-positive OLGs of total Olig2-positive cells in (h) (n = 8 for 3 days, n = 10 for 6 days, at least 150 cells analysed per independent experiment) and the number of MBP-positive cells that elaborate myelin membranes in (I) (n = 10, 6 days). Note that after 3 days of differentiation wmOLGs are larger and morphologically more complex than gmOPCs. In addition, wmOPCs show an accelerated differentiation, while the number of MBP-positive cells bearing myelin membranes at day 6 is similar. Bars represent means. Error bars show the standard error of the mean. Statistical analyses were performed using a paired two-sided t-test (*p < 0.05, ***p < 0.001). Scale bar is 50 µm.