13287_2018_779_MOESM2_ESM.tif (9.53 MB)

Additional file 2: Figure S1. of One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system

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posted on 26.02.2018 by Methichit Wattanapanitch, Nattaya Damkham, Ponthip Potirat, Kongtana Trakarnsanga, Montira Janan, Yaowalak U-pratya, Pakpoom Kheolamai, Nuttha Klincumhom, Surapol Issaragrisil
Showing characterization of iPSCs derived from skin fibroblasts of a patient with hemoglobin E/beta-thalassemia. (a) Pluripotent gene expression of wild-type human dermal fibroblasts (wt-HDFs), parental human dermal fibroblasts (Eβ-HDFs) and Eβ-iPSCs compared with hESC line Chula2.hES, analyzed by RT-PCR. (b) Immunofluorescent staining shows expression of pluripotent markers NANOG, OCT4, SSEA-4, TRA-1-60 and TRA-1-81 in the Eβ-iPSC1 and Eβ-iPSC2 cells. Scale bars = 100 μm. (c) Immunofluorescent staining shows expression of lineage markers NESTIN (ectoderm), AFP (endoderm) and SMA (mesoderm) of differentiated embryoid bodies generated from the Eβ-iPSC1 and Eβ-iPSC2 cells. Scale bars: for NESTIN and SMA = 100 μm; for AFP = 50 μm. (d) Hematoxylin and eosin (H&E) staining of teratomas derived from the Eβ-iPSC2 cells at 8 weeks post implantation into nude mice. Teratomas contained tissues derived from three embryonic germ layers, sebaceous tissue (ectoderm), cartilage (mesoderm) and gut-like epithelium (endoderm). Scale bars = 100 μm. (e) Representative karyotypic analysis of the Eβ-iPSC2 cells at passage 19 shows normal karyotype (46, XY) (TIFF 9760 kb)


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