Characterization of Human Induced Pluripotent Stem Cell Lines Derived from Neonatal and Adult Human Dermal Fibroblasts

Somatic cells can be reprogrammed to Induced Pluripotent Stem Cells (iPSCs) by expressing four transcription factors, Oct4, Sox2, Klf4 and c-Myc. Co-expressing Rarg (retinoic acid receptor gamma) and Lrh-1 (liver receptor homolog 1, Nr5a2) with the four factors greatly accelerated reprogramming so that reprogramming of mouse embryonic fibroblast cells (MEFs) to ground state iPSCs requires only four days’ induction of these six factors. The six-factor combination readily reprogrammed primary human neonatal and adult fibroblast cells to exogenous-factor-independent iPSCs, which resembled ground state mouse ES cells in growth properties, gene expression and signalling dependency. Our findings demonstrate that signalling through RARs has critical roles in molecular reprogramming and the synergistic interaction between Rarg and Lrh1 directs reprogramming towards ground state pluripotency. SH-iPS20-1, -3 and -9 lines were derived from HDFa (439656), SH-iPS24-1, -2 and -11 lines were derived from HDFa (709590), and SH-iPS28-23, -25 and -27 lines were derived from HDFn (617769). Parental HDF lines were used as control. SH-iPS15-5 and -10 lines were derived from HDFn (617769, Invitrogen). H1 hESC were obtained from Wicell (Madison, USA). SH-iPSCs were cultured in both 2i+LIF and FGFconditions. Expression pattern of SH-iPSCs was compared to H1 hESC cultured in FGF conditions. S1, S2 and S3 are three indpendent subcultures of a cell line.