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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Biphasic modulation of insulin signaling enables highly efficient hematopoietic differentiation from human pluripotent stem cells

Fig. 2

Biphasic modulation of insulin signaling promoted highly efficient HSPC differentiation. a Schematic stepwise induction of hematopoietic progenitors from hPSCs. b Representative images of stages I–III of hematopoietic differentiation. “Grape-like” clusters at differentiation day 6 indicated by arrows. Scale bars, 50 μm. c Time-lapse images of day 6 differentiating cells. Cells undergoing endothelial-to-hematopoietic transition indicated by arrowheads. Scale bars, 50 μm. d Representative immunostaining images of day 8 (i–iii) and day 13 (iv) cells for CD34 and VE-cadherin (both green), and CD43, CD31 and CD45 (all red). (i–iii) Grape-like clusters coexpressed CD43, CD34, VE-CAD and CD31(white arrow). (iv) CD45+CD43low and CD45lowCD43+ indicated by arrows and arrowheads respectively. Scale bars, 50 μm. e Strategy for biphasic insulin treatment. Cells differentiated in B27 without insulin (B) or B27 with insulin (BI). Insulin was added at different stage as indicated. f Representative flow cytometry results of surface markers CD31, CD43 and CD34 at day 5, day 6 and day 8. N > 3 independent biological replicates. g Bar graph of percentage of CD31+CD34+, CD31+CD34 and CD31CD34+ cells in B + B and B + BI groups on day 8. N > 3 independent biological replicates. h Bar graph of percentage of CD43+CD34+, CD43+CD34 and CD43CD34+ cells in B + B and B + BI group on day 8. N > 3 independent biological replicates. HE hemogenic endothelium, hPSC human pluripotent stem cell, HSPC hematopoietic stem/progenitor cell, ME mesoderm

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