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. 2013 Jan 31;8(1):e55226. doi: 10.1371/journal.pone.0055226

Figure 9. Generation of hiPSCs from DMCs with non-conditioned hESC medium on PCM-DM.

Figure 9

A) Morphology and Alkaline phosphatase (ALP) staining of iPS-DMC75-PCMDM. P, passage number. Scale bar = 500 µm. B). Quantitative RT-PCR analysis for the mRNA copy number of four transgenes (OCT4, SOX2, KLF4, c-MYC). All the transgenes were silenced in iPS-DMC75-PCMDM. Data are presented as the mean ± SD. *: not detected. C) Immunocytochemistry for NANOG (red) and OCT4 (green) expression in iPS-DMC75-PCMDM. Scale bar = 200 µm. D) Flow cytometry analysis for hESC-specific surface antigens (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) at passage 11. E) Global gene expression analysis of iPS-DMC75-PCMDM (passage 12), early and late passages of clones 1 and 2, KhES1, and 201B7. Scatter plots and Pearson’s coefficient are shown. The diagonal lines indicate 3-fold changes in gene expression levels. Plus (“+”) symbols indicate stem-cell marker genes suggested by the International Stem Cell Initiative [31], and such genes outside the 3-fold change lines are shown in red text. Clone 1, iPS-DMC72-PCMDM01; Clone 2, iPS-DMC72-PCMDM02. F) Embryoid bodies (EBs) on day 8 derived from iPS-DMC75-PCMDM (passage 12). Scale bar = 500 µm. G) Quantitative RT-PCR analysis. Left, Expression levels of genes for the undifferentiated state in EBs relative to hiPSCs before differentiation. Right, Expression levels of lineage-specific genes in EBs relative to differentiated 201B7. Data are presented as the mean ± SD.