Figure 1

Early passage mixed glial cultures are composed of astrocytes, microglia, fibroblasts and pericytes while later passages are primarily pericytes and/or fibroblasts. Cells from at least three individual cases at passage 2 were stained by immunocytochemistry for the cell specific markers fibronectin (A and D) (green), prolyl-4 hydroxylase (P4H) (B and E) (green), and PDGFRβ (C and F) (green) and co-labelled with NG2 (A-C) (red) or αSMA (D-F) (red), or glial fibrillary acidic protein (GFAP) (green) and CD45 (red) (G). Bottom images show overlay with Hoechst (blue) for all images. Scale bar = 100 μM. Percentages of αSMA, PDGFRβ or αSMA/PDGFRβ positive cells from passage 2 or 5 from three separate cases were quantified from triplicate wells, four images/well (H). PDGFRβ, platelet-derived growth factor receptor-beta; αSMA, alpha smooth muscle actin.