Permissiveness to form pluripotent stem cells may be an evolutionarily derived characteristic in Mus musculus

Sci Rep. 2018 Oct 2;8(1):14706. doi: 10.1038/s41598-018-32116-8.

Abstract

Mus musculus is the only known species from which embryonic stem cells (ESC) can be isolated under conditions requiring only leukemia inhibitory factor (LIF). Other species are non-permissive in LIF media, and form developmentally primed epiblast stem cells (EpiSC) similar to cells derived from post-implantation, egg cylinders. To evaluate whether non-permissiveness extends to induced pluripotent stem cells (iPSC), we derived iPSC from the eight founder strains of the mouse Collaborative Cross. Two strains, NOD/ShiLtJ and the WSB/EiJ, were non-permissive, consistent with the previous classification of NOD/ShiLtJ as non-permissive to ESC derivation. We determined non-permissiveness is recessive, and that non-permissive genomes do not compliment. We overcame iPSC non-permissiveness by using GSK3B and MEK inhibitors with serum, a technique we termed 2iS reprogramming. Although used for ESC derivation, GSK3B and MEK inhibitors have not been used during iPSC reprogramming because they inhibit survival of progenitor differentiated cells. iPSC derived in 2iS are more transcriptionally similar to ESC than EpiSC, indicating that 2iS reprogramming acts to overcome genetic background constraints. Finally, of species tested for ESC or iPSC derivation, only some M. musculus strains are permissive under LIF culture conditions suggesting that this is an evolutionarily derived characteristic in the M. musculus lineage.

Publication types

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

MeSH terms

  • Animals
  • Cells, Cultured
  • Cellular Reprogramming / drug effects
  • Cellular Reprogramming / genetics*
  • Culture Media / metabolism
  • Culture Media / pharmacology
  • Evolution, Molecular*
  • Female
  • Fibroblasts
  • Glycogen Synthase Kinase 3 beta / antagonists & inhibitors
  • Glycogen Synthase Kinase 3 beta / metabolism
  • Induced Pluripotent Stem Cells
  • Leukemia Inhibitory Factor / metabolism
  • MAP Kinase Kinase Kinases / antagonists & inhibitors
  • MAP Kinase Kinase Kinases / metabolism
  • Male
  • Mice / genetics*
  • Mice, Inbred Strains / genetics
  • Mouse Embryonic Stem Cells
  • Primary Cell Culture
  • Protein Kinase Inhibitors / pharmacology
  • Sex Factors
  • Transcription, Genetic*

Substances

  • Culture Media
  • Leukemia Inhibitory Factor
  • Lif protein, mouse
  • Protein Kinase Inhibitors
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • MAP Kinase Kinase Kinases