File:Cord blood induced stem cells 01.jpg

Generation of non-viral induced pluripotent stem cells (hiPSC) from CD34+ cord blood progenitors

(A) Representative immunocytochemistry of pluripotency markers POU5F1 (OCT4), NANOG, TRA-1-81, and SSEA4 in hiPSC line CBiPSC6.2 after >20 passages. (B) Representative hematoxylin and eosin staining of teratoma sections derived from CBiPSC6.2 after >20 passages demonstrating ectodermal, endodermal and mesodermal lineage differentiation. All CBiPSC clones in these studies formed similar teratomas. (C) Real-time RT-PCR studies p15 CBiPSC lines for endogenous pluripotency genes using primers that distinguish endogenous expression from transgenes (see Methods). (D) The presence of plasmid transgene sequences examined by PCR at p11 in CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (lanes 3–6, respectively) and negative control H9 hESC (p48) (lane 1) compared to positive control early cultures from p2 (lane 2). RT-PCR analysis of selected plasmid sequences in p11 CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (lanes 8–11, respectively) and negative control H9 hESC (lane 7) and p2 cultures (lane 12). (E) Genomic Southern blot analysis for episomal vector backbone integration in lines CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (p15) (lanes 2–5, respectively), H9 hESC (p55) (lane 1). Combination 6 episomal vector DNA was diluted as positive control to the equivalents of 0.4 and 4 integrations per haploid genome (0.4× and 4×). L: 1 kb plus ladder. These studies were also conducted for non-viral adult fibroblast-derived hiPSC lines iPSCWT2 and iPSCWT4 with similar results (Machairaki et al., in preparation).

Original image name: Figure 2. doi:10.1371/journal.pone.0018293.g002 (cropped A-c from full figure)

Reference

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3072973/

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0018293

Copyright: © 2011 Burridge 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 credited.