Generation of iPSCs from non-cryoprotected dura. Scale bars are 1 cm for (A) and 100 μm for (B-M). (A) Gross examination of a coronal right hemi-section from the brain of a sporadic AD patient (case ASC7) illustrates atrophy and ventricular dilatation. Immunohistochemical staining for amyloid-β (Aβ) peptide and hyper-phosphorylated tau confirms pathological accumulation of amyloid plaques and neurofibrillary tangles (insets). Scale bar is 1 cm. (B) Fibroblast-like outgrowths from the thawed archived dura from the same subject, 18 days post plating. (C-N) Characterization of a representative iPSC clone (clone 4) derived ASC7D-AD. (C-E) Immunofluorescence staining using antisera targeting (C) Nanog (green), Tra160 (red); (D) Sox2 (green), SSEA4 (red); (E) Oct4 (green) alkaline phosphatase (red) confirms pluripotency. Nuclei are counterstained with Hoechst 33342 (blue). (F-G) Undirected EBs were cryosectioned and immunostained for the three developmental germ layers: endoderm (Sox17), mesoderm (SMA), and ectoderm (Tuj1). (H-J) Teratomas were sectioned and hematoxylin and eosin stained, and show evidence of the presence of the three developmental germ layers as indicated. (K-L) Nanostring analysis for endogenous stem cell genes (K), and shutoff of Sendai transgenes (L). (M) Immunofluorescence staining using antisera targeting neuron-specific class III β-tubulin (Tuj1, green) and the neural progenitor marker paired box 6 (PAX6, red) demonstrates directed neuronal differentiation (21 days). Nuclei are counterstained with DAPI (blue). (N) This iPSC line displays a normal female karyotype.