Journal: Nature Communications

Article Title: Parallel derivation of isogenic human primed and naive induced pluripotent stem cells

doi: 10.1038/s41467-017-02107-w

Figure Lengend Snippet: Direct reprogramming of somatic cells into hiNPSCs. a Direct generation of isogenic naive and primed hiPSCs. Fibroblasts were transduced with 3 Sendai viruses expressing a polycistron KLF4/OCT4/SOX2, MYC and KLF4 at a ratio of 5:5:3, respectively. Cells were split on feeders at day 7, and placed in the indicated media at day 9. Scale bar = 100 µm. b Summary of lines generated for this study in primed (KSR+FGF2, yellow) or naive culture media (RSeT, blue or T2iLGö, pink) originated from 5 different donors. c Different pluripotent states are induced depending on culture media. Transcriptomes of hiPSCs and hiNPSCs, control primed hESC lines H1 and H9 or the naive hESC line HNES1 were analyzed by PCA. Symbols represent donor lines, and size of the symbols represents the passage. Arrows have been drawn to highlight the reprogramming trajectories. d T2iLGö hiNPSCs are the closest to human epiblast cells. PCA of single-cell RNA-seq data sets from preimplantation embryo samples , compared to primed hPSCs from ref. and to primed/naive hiPSCs/hESCs from this study

Article Snippet: Primed hiPSCs from L7 human adult dermal fibroblasts extracted from a healthy male aged 51 years (Lonza cat. no. CC-2511 Lot 0000293971) were also used in this study. hESC lines H1 (WA01 Lot WB0111) and H9 (WA09 Lot WB0090) were obtained from the WiCell Research Institute, under authorization RE13-004 from the French embryo research oversight committee, Agence de la Biomédecine.

Techniques: Transduction, Expressing, Generated, RNA Sequencing Assay