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. 2015 Jul 20;125(8):3103–3116. doi: 10.1172/JCI80924

Figure 1. Impaired granulopoietic differentiation of hematopoietic progenitors derived from SCN patient iPSCs.

Figure 1

(A) Colony forming cell assay of the hematopoietic progenitors derived from healthy donor (control 12 and control 13) and ELANE-mutant SCN iPSCs (SCN15 and SCN14). Cells were cultured in methyl cellulose semisolid medium with cytokines, and the myeloid (CFU-G, CFU-GM, CFU-M) and mix colonies were scored at day 14. (B) Schematic diagram of granulopoietic differentiation of iPSC-derived hematopoietic progenitors. (C) Top: Quantitative analyses of the granulopoietic differentiation of control iPSC–derived (control 12–derived) hematopoietic progenitors in liquid culture with myeloid differentiation cytokines. Bottom: Wright-Giemsa–stained cytospin at the end of the culture. (D) Top: Quantitative analyses of the granulopoietic differentiation of control iPSC–derived (control 13–derived) hematopoietic progenitors in liquid culture with myeloid differentiation cytokines. Bottom: Wright-Giemsa–stained cytospin at the end of the culture. (E) Top: Quantitative analyses of the granulopoietic differentiation of hematopoietic progenitors derived from SCN15 iPSCs in liquid culture condition with myeloid differentiation cytokines. Bottom: Wright-Giemsa–stained cytospin at the end of the culture. (F) Top: Quantitative analyses of the granulopoietic differentiation of hematopoietic progenitors derived from SCN14 iPSCs in liquid culture with myeloid differentiation cytokines. Bottom: Wright-Giemsa–stained cytospin at the end of the culture. (G) FACS-based ROS-generating-activity analyses of promyelocytes derived from control and SCN iPSCs. Scale bars: 10 μm. ProM, promyelocytes; My, myelocytes; MM, metamyelocytes; Neutr, neutrophils; Monos, monocytes. Data are presented as mean ± SD of individual values out of 4 or 5 independent experiments. Cont, control.