Fig. 6
R-spondin2 ultimately augments the production of functional platelets from hPSCs. a Comparative analysis of the number of cobblestone-like HPCs after mechanical detachment from individual culture dishes of mAGM-S3 co-culture with or without R-spondin2 (20 ng/mL) treatment at day 1. Results are shown as means ± SD (n = 3). *P < 0.05. b Representative morphologies of large cells and proplatelets at day 6 of MK culture. c Quantitative analysis of CD41a+CD42b+ MKs generated from each H1 cells. Results are shown as means ± SD (n = 3). *P < 0.05. d Representative morphologies of multinuclear MKs by MGG (May-Grunwald-Giemsa) Staining at day 3 of megakaryocytic differentiation (scale bar, 20 mm). e Quantitative analysis of CD41a+CD42b+ PLPs generated from each H1 cells. Results are shown as means ± SD (n = 3). **P < 0.01. f Representative immunofluorescence images of hPSC-derived PLPs bound to immobilized fibrinogen with F-actin and CD41 stained in the absence (top) or the presence (bottom) of 1 U/mL thrombin. Scale bar = 5 μm. g Representative immunofluorescence images showing aggregation in a mixture of 2 × 105 Calcein-AM (red)-labeled hPSC-derived PLPs and 2 × 107 blood platelets. Scale bar = 5 μm. h Left panel: representative flow cytometry analysis of P-selectin (CD62P) in gated CD41a+ PLPs. Right panel: median fluorescence intensity analysis of CD62P in gated CD41a+ PLPs. Results are shown as means ± SD (n = 3). NS, not significant