Fig 2: Characteristics of the isolated cells. A The morphology of passage 1~3 (P1/P2/P3) cells from rats. B The adipogenic induction and oil red O staining. C The analysis of the cell cycle of the isolated cells by flow cytometry. D The expression of surface markers of the isolated cells; Typical hematologic lineage CD29 and CD90 were expressed in cells, while CD45 and CD34 were not. Scale bars, 50 μm

Fig 3: EPO treatment enhances the production of CD34+ cells and mobilizes CD34+/VEGF-R2+ cells from bone marrow to peripheral blood.(A–D) Flow cytometric analysis of the populations of CD34+ and CD34+/VEGF-R2+ cells isolated from bone marrow (BM) and peripheral blood (PB). CNTX = non-dystrophic control rats without EPO treatment, CTX = non-dystrophic control rats receiving EPO treatment, DNTX = RCS rats without EPO treatment, DTX = RCS rats receiving EPO treatment. Box plots display the 25th percentile, median (50th percentile), 75th percentile as well as the minimum and maximum number of cells analyzed. *P<0.01, EPO treated group vs not treated group; n = 7∼10/group.

Fig 4: Lineage identification and characterization of rat MSCs. (A) Expression of rat MSC markers (CD44 and CD90.1), endothelial cell marker (CD31), leukocyte marker (CD45), and hematopoietic stem cell marker (CD34) on rat BMSCs. (B) Alizarin Red S staining of rat BMSCs detected after culture for 21 days in osteogenic medium or basal medium. (C) Chondrogenic differentiation potential of the micromass of rat BMSCs in vitro. Alcian blue staining of cells was detected after culture for 14 days in chondrogenic medium or basal medium. (D) Adipogenic differentiation potential of rat BMSCs in vitro.Note: Oil Red O staining of cells was detected after culture for 14 days in adipogenic medium or basal medium.Abbreviations: MSC, mesenchymal stem cell; BMSCs, bone marrow mesenchymal stem cells.

Fig 5: Scatter diagram of leukocyte measured by flow cytometry in the peripheral blood of rat. Whole fresh blood samples anti-coagulated by EDTA.2Na were incubated with PE-conjugated anti-CD34 antibody and FITC-conjugated anti-vWF antibody. Then hemolysin was dissolved in distilled water, and applied to cells at the final concentration 10% (v/v). (A) The forward-scatter (FSC) and side-scatter (SSC) diagram, Cells in gate represent the mononuclear cells; (B) The fluorescence intensity scatter diagram. MNCs that were positively identified as CEPC by immunofluorescent staining for the PE+/FITC+ or CD34+/vWF+ (P2-Q2) represent EPC.