(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.