Characterization of X-CGD iPSC line iXC1. Line iXC1 (A) exhibits characteristic human ESC-like morphology in bright field (BF) microscopy (top left) and expresses pluripotency markers that include alkaline phosphatase (AP) activity (top right) and fluorescence antibody staining for OCT4, NANOG, TRA-1-60, and SSEA-4 (left panels in green and red). Blue staining (right panels) are 4′-6-Diamidino-2-phenylindole (DAPI) staining of nuclei (bar = 100 μm); (B) has the CYBB 458T→G mutation; (C) has normal male karyotype; (D) forms embryoid bodies in vitro shown by BF (top left) microscopic appearance, and by expression of β-tubulin class III neuroectoderm marker (top right), smooth muscle actin (SMA) mesoderm marker (bottom left), and α-fetoprotein (AFP) endoderm marker (bottom right); (E) forms teratomas in vivo in NSG mice that include all 3 germ layers, identified here as neural rosette ectoderm (top), cartilage mesoderm (middle), and respiratory epithelial endoderm (bottom). Not shown is that RT-PCR analysis of cDNA generated from the iXC1 teratoma confirmed presence of mRNA specific for human PAX6 (ectoderm), CD34 (mesoderm), and α-feto-protein (endoderm).