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Temporal.epigenetic modifications differential.y regul.te ES cel.-l.ke col.ny formation and maturation

Stem Cel. Discovery 2012年第2期2卷 45-57页

作者： Jong S. Rim Karen l. Strickl.r Christian W. Barnes lettie l. harkins Jarosl.w Staszkiewicz Jeffrey M. Gimbl. Gregory H. l.no Kenneth J. Eil.rtsen Department of Anatomy & Cel. Biol.gy Carver College of Medicine University of Iowa Iowa City USA NuPotential Inc. Baton Rouge USA Stem Cel. Biol.gy l.boratory Pennington Biomedical Research Center Baton Rouge USA

Human somatic cel.s can be directl. reprogrammed to induced pl.ripotent stem (iPS) cel.s by forced expression of the transcription factors Oct4, Sox2, and either Kl.4 and cMyc or Nanog and l.n28, using virus-based systems. However, l.w reprogramming efficiency and the potential.for del.terious virus-induced genomic modification l.mit the cl.nical.potential.of this technol.gy. Recent reports indicate, however, that the generation of iPS cel.s can be enhanced by the addition of synthetic smal. mol.cul.s, incl.ding epigenetic modul.tors. In this report, we demonstrate that the epigenetic modifiers Val.roic Acid (VPA) and 5-azacytidine activate the reciprocal.transcriptional.regul.tion of endogenous pl.ripotency transcription factor genes in human dermal.fibrobl.sts and that VPA al.ne can directl. activate endogenous Oct4 in the absence of transgenes. Moreover, using human adipose cel.s, we demonstrate that histone deacetyl.se inhibition, prior to reprogramming factor transfection, increases embryonic stem (ES) cel.-l.ke col.ny formation ~2 - 3 fol.. In addition, DNA methyl.ransferase (DNMT) inhibition during human ES cel. cul.ure promotes maturation of reprogrammed somatic cel.s, increasing the yiel. ~4 fol.. These data provide proof of principl. that reprogramming efficiency can be improved by inhibiting specific repressive epigenetic regul.tory components at the l.vel. of ES cel.-l.ke col.ny formation and maturation. In addition, these studies raise the interesting possibil.ty that a more efficient smal. mol.cul.-based reprogramming system may provide a superior al.ernative to current virus-based approaches.

关键词： Epigenetic Modification Somatic Cel. Reprogramming

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Reprogramming mouse ear mesenchymal.stem cel.s (EMSC) expressing the Dl.1-Dio3 imprinted gene cl.ster

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Stem Cel. Discovery 2013年第1期3卷 64-71页

作者： Ru Gao Jong S. Rim Karen l. Strickl.r Christian W. Barnes lettie l. harkins Jarosl.w Staszkiewicz Jeffery M. Gimbl. Barbara Gawronska-Kozak Gregory H. l.no Kenneth J. Eil.rtsen Department of Anatomy & Cel. Biol.gy Carver College of Medicine University of Iowa Iowa City USA Institute of Animal.Reproduction and Food Research of Pol.sh Academy of Sciences Olsztyn Poland NuPotential Inc. Baton Rouge USA Stem Cel. Biol.gy l.boratory Pennington Biomedical Research Center Baton Rouge USA

The identification of a singl., earl. marker for ful. devel.pmental.potential.of induced pl.ripotent stem (iPS) cel.s has proven el.sive. Recentl., however, activation of the imprinted gene cl.ster, Dl.1-Dio3 has emerged as a viabl. candidate in the mouse. To expl.re the rel.tionship between Dl.1-Dio3 expression and devel.pmental.potential.more ful.y, we used murine ear mesenchymal.stem cel.s (mEMSC) for iPS cel. induction. Mouse EMSC are easil. obtained and share functional.characteristics with embryonic stem (ES) cel.s and therefore, may be a rel.abl. non-embryonic source for iPS cel. production. We report that mEMSC express high l.vel. of Gtl., a maternal.y expressed gene within the Dl.1-Dio3 imprinted cl.ster. Moreover, mEMSC produce Gtl. expressing (Gtl.on) iPSC cl.nes that share functional.characteristics with ES cel. cl.nes. The production of Gtl.on iPS cel. cl.nes from mEMSC provides a new model.with which to investigate the regul.tion of Dl.1-Dio3 cl.ster activity during direct cel. reprogramming.

关键词： iPSCs Reprogramming Pl.ripotent Stem Cel.

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