Culture and Differentiation of Oligodendrocyte Progenitor Cells (OPCs).

<p>A) OPCs cultured for 3 days after extraction on poly-L-lysine coated culture dishes. Numerous astrocytes are present. B) OPCs after passaging and expanding in the presence of FGF2 and PDGF-AA, to induce proliferation and maintenance of the OPCs. C) OPCs cultured in differentiation inducing medium for 7 days. Mature oligodendrocytes appear in the culture. D) OPCs cultured for 7 days in the differentiating medium and FGF8. All images taken at 100X magnification. E–G) Immunocytochemistry of OPCs in undifferentiating medium (E), differentiation inducing medium (F), and in the presence of FGF8 (G), staining for Olig2 (in green) and DAPI for nuclear staining (in blue). H) Histogram presenting the percentage of Olig2+ cells in the various cultures observed in E–G. Percentages are with respect to the total number of cells, calculated by DAPI staining. I–K) Immunocytochemistry of NG2+/Brdu+ cell in undifferentiating medium (I), differentiation inducing medium (J), and in the presence of FGF8 (K). L) Histogram presenting the percentage of NG2+/BrdU+ cells in the different cultures, with respect to the total number of cells. *p<0.05 (paired t-test), n = 4. Scale Bar indicates 150 µm for A–D and 100 µm for E–G and I–K.