A murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells

Cell Stem Cell. 2010 Jun 4;6(6):535-46. doi: 10.1016/j.stem.2010.05.003.

Abstract

Murine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human iPSC reprogramming in the presence of LIF yields human stem cells that display morphological, molecular, and functional properties of murine ESCs. We termed these hLR5 iPSCs because they require the expression of five ectopic reprogramming factors, Oct4, Sox2, Klf4, cMyc, and Nanog, to maintain this more naive state. The cells are "metastable" and upon ectopic factor withdrawal they revert to standard human iPSCs. Finally, we demonstrate that the hLR5 state facilitates gene targeting, and as such provides a powerful tool for the generation of recombinant human pluripotent stem cell lines.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation / metabolism
  • Cell Dedifferentiation / drug effects
  • Cell Dedifferentiation / genetics
  • Cell Line
  • Embryonic Stem Cells / metabolism*
  • Embryonic Stem Cells / pathology
  • Fibroblast Growth Factor 2 / metabolism
  • Gene Transfer Techniques*
  • Genetic Therapy / methods
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism*
  • Induced Pluripotent Stem Cells / pathology
  • Kruppel-Like Factor 4
  • Leukemia Inhibitory Factor / pharmacology*
  • Mice
  • Recombination, Genetic / genetics
  • Sequence Homology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Antigens, Differentiation
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Leukemia Inhibitory Factor
  • Lif protein, mouse
  • Transcription Factors
  • Fibroblast Growth Factor 2

Associated data

  • GEO/GSE21792