Immunophenotypic characterization and expansion of cultured cord-blood MSCs. Culturing cbMSCs from cbMNCs resulted in enrichment of CD73+CD166+CD90+CD45-CD34-HLA-DR- cells closely resembling the phenotypes of MSC proposed by the International Society for Cellular Therapy (ISCT), 2006 [72]. Representative flow-cytometry contour plots are shown to illustrate expression of individual phenotypic markers in cbMNCs and cbMSCs (A). In each plot, percentage of cells positive for a given marker is shown on the right, and percentage of cells negative for the same marker is shown on the left. Relative to cbMNCs, cbMSCs had enriched expression of CD73, CD166 (P < 0.0001), CD90 (P < 0.0004), and decreased expression of HLA-DR (P < 0.0001), CD45 (P < 0.01), and CD19 (P < 0.005). cbMSCs also had relatively higher expression of HLA-ABC and lower expression of CD34 and CD133, although the difference was not significant (B). Spindle-shaped, fibroblast-like, flat, adherent cells are shown across various passages of cbMSC culture (magnification 10×) (C). Comparative morphologic representation of 10th passage cbMSCs and commercially obtained cmMSCs is shown in (D). Student t test, n = 4 for each group, (**) is P < 0.01, (***) is P < 0.001, and (ns) is nonsignificant. Data represent means ± SD.