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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Identification of circulating CD31+CD45+ cell populations with the potential to differentiate into erythroid cells

Fig. 1

Flow cytometric characterization of circulating CD31highCD45+ and CD31lowCD45+ cells in human peripheral blood. a Strategy for the isolation of Lin cells from “whole blood” for sequential cytokine stimulation and bone marrow mesenchymal stromal cell co-culture to promote erythroblast differentiation. b Represenative flow cytometric analysis of fresh adult human Lin CD31highCD45+ and CD31lowCD45low cell populations from enriched hematopoietic Lin cells from peripheral blood. c Frequency of CD31+ and CD45+ cells (bar graph). The total percentage of CD31highCD45+ and CD31lowCD45low cells and of the control CD31CD45+ population (from the entire Lineage-negative cell population) are displayed as mean of n = 3 independent experiments ± SD. d Mean percentage of CD117+, CD34+, CD150+, and CD133+ cells ± SD are shown; n = 3 independent experiments. e Representative plots showing the distribution of CD31highCD45+ and CD31lowCD45low cell populations and immunophenotypic changes after 3 days of cytokine stimulation. f Cumulative data are shown as means ± SD of 3 independent experiments in the bar graph. Suspended and semi-adherent hematopoietic cells were collected for flow cytometry analysis. g Mean percentage of CD117+, CD34+, and CD150+ cells ± SD are shown; three independent experiments. h Mean percentage of ±SD CD34+CD117+ cells detected on day 0 and after 3 days of cytokine stimulation ± SD are shown; three independent experiments. i Representative flow cytometric analysis from the entire Lineage-negative population after 3 days of cytokine stimulation and 15 days of co-cultivation with MSCs. The cells lost CD31highCD45+ and CD31lowCD45low expression and acquired CD235b+ expression erythroid cells; n = 3 independent experiments. j Cell yields at the indicated time points (left panel). k The left panel shows representative fluorescence imaging (CD31 and CD45 markers) from the entire Lineage-negative cell population at day 0 of culture and bright-field microscopy images (original magnification 40×) after cytokine stimulation for 3 days; cells were stained with Wright-Giemsa at day 18 (after the 3 days of cytokine stimulation and the 15 days of co-culture on MSCs). Scale bars, 20 μm. The right panel shows representative images of basophilic erythroblasts (Baso), polychromatic erythroblasts (Poly), orthochromatic normoblasts (Ortho), on day 18 (3 + 15 days) after erythroid differentiation with cytokines and bone marrow mesenchymal stem cell co-culture; n = 3 independent experiments. Data are shown as means ± SD

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