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. 2012 Dec 13;7(12):e48677. doi: 10.1371/journal.pone.0048677

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© 2012 Wakao et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A–D) Immunocytochemistry for (A) Oct3/4, (B) Nanog, (C) Sox2, (D) TRA-1-81 in iPS cell colony G. Scale bar = 100 µm. (E, F) EBs generated from colony G were plated on gelatin coated dishes containing DMEM/F12 medium supplemented with 20% knockout serum replacement. After 10 days, cell differentiation was confirmed by immunocytochemistry for mesodermal (smooth muscle actin; SMA) (E), endodermal (alpha-fetoprotein; alpha-FP) (E) and ectodermal markers (neurofilament; NF) (F). Scale bar = 100 µm. (G) RT-PCR of differentiation markers in undifferentiated iPS cell colony G (Undifferentiation) and embryoid bodies derived from iPS cell colony G. Differentiation). (H–K) Hematoxylin and eosin staining of teratoma formed by transplantation of iPS cell colony G into immunodeficient mice testis. (H), Low magnification of the formed teratoma (12 weeks after injection). Endodermal (I), mesodermal (J) and ectodermal (K) tissue were observed in the teratoma.