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. 2015 Nov 11;52:252–264. doi: 10.1007/s11626-015-9968-x

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© The Author(s) 2015

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Figure 1. — Integration-free hiPS cell generation from DPCs in serum- and feeder-free culture conditions using SeVdp. (A) Genome structure of defective persistent Sendai virus (SeVdp) vector. SeVdp has mutations in the L and P genes, which are responsible for low cytotoxicity and for defective induction of IFN-b. The M, F and HN genes are deleted and replaced with genes interest (KOSM). SeVdp-iPS was installed with Klf4, Oct4, Sox2, c-Myc cDNA on a single vector. (1) Insertion of Gene-End signal, (2) Mutation in P gene (P517H), (3) Mutation in L gene (V981I, S1088A, C1207S, V1618L) (4) Deletion of M, F and HN genes, and installation of exogenous genes. (B) Time schedule of hiPS cell generation. Day −7∼0 DPCs were cultured in RD6F serum-free medium on type I collagen coated dish. Day 0∼1 SeVdp (KOSM) transduction (Oct4, Sox2, Klf-4, c-Myc) with hESF9 medium. Day 1 re-seeding on fibronectin-coated plate with hESF9 medium. Day 2∼30 exchange medium every other day. (C) Phase contrast images of iPSCs derived from normal DPCs (SeV-M-iPS and SeV-H-iPS). (upper panels) SeV-M-iPS clone1 at passage 11 on fibronectin-coated dish with hESF9 medium. (lower panels) SeV-H-iPS-clone25 at passage 50, clone35 at passage 60 and passage 87 on fibronectin-coated dish with hESF9 medium. Bars indicate 200 μm. DPC-H-derived iPS cells generated with Sendai virus were designated SeV-H-iPS and DPC-M-derived iPS cells were designated SeV-M-iPS.