Characterization of iPSCs from patient with CCALD. (A) Representative images of alkaline phosphatase (AP) staining and pluripotency protein biomarker immunostaining analysis of childhood cerebral adrenoleukodystrophy (CCALD) patient induced pluripotent stem cell (iPSCs) are presented. (B) Representative CCALD iPSCs were subject to in vitro differentiation (Materials and methods) and cell lineages derived from all three germ layers were identified by immunostaining, as depicted. Antibodies against Tuj1, αSMA and AFP were used to identify cells of ectodermal, mesodermal, and endodermal origin, respectively. (C) Histological analysis of teratomas derived from representative CCALD iPSC clones from two patients is provided. Evidence of cell lineages derived from all three germ layers is provided. C, Cartilage tissue; G, Glandular tissue; N, Neural rosettes; P, pigment neuroepithelium. Scale bar = 50 μm.