Fig. 4

Brain PCs serve as multipotent VSCs following ischemia. Cells were isolated from ischemic areas, and PDGFRβ⁺ iPCs were selectively collected using MACS (a). By immunohistochemistry, MACS-sorted cells expressed both PDGFRβ and nestin (PDGFRβ (b, c red), nestin (c green), DAPI (b, c blue)). After incubation in a floating culture (d), PDGFRβ⁺ iPCs formed clusters (e). The cells in these clusters expressed nestin (PDGFRβ (f, g green), nestin (f, h red), DAPI (f–h blue)) and PC cell marker proteins that included NG2 (NG2 (i green), DAPI (i: blue)), and αSMA (αSMA (j green), DAPI (j blue)). PCR analysis confirmed that the clusters expressed PC markers (PDGFRβ, NG2, and αSMA). In addition, these cells expressed various stem and undifferentiated cell markers, including nestin, c-myc, Klf4, and Sox2 (k). The PDGFRβ⁺ iPCs subsequently differentiated into various cell types, including Tuj1⁺ neuronal cells (l), osteopontin⁺ osteoblasts (m), FABP4⁺ (n), and Oil red⁺ adipocytes (o). Scale bars = 50 μm (b, e, f, i, j), and 20 μm (l, m, n, o) FABP4 fatty acid binding protein 4, MACS, magnetic cell sorting, PDGFRβ platelet-derived growth factor receptor-β, VSCs vascular stem cells