Derivation of hypermethylated pluripotent embryonic stem cells with high potency

Cell Res. 2018 Jan;28(1):22-34. doi: 10.1038/cr.2017.134. Epub 2017 Oct 27.

Abstract

Naive hypomethylated embryonic pluripotent stem cells (ESCs) are developmentally closest to the preimplantation epiblast of blastocysts, with the potential to contribute to all embryonic tissues and the germline, excepting the extra-embryonic tissues in chimeric embryos. By contrast, epiblast stem cells (EpiSCs) resembling postimplantation epiblast are relatively more methylated and show a limited potential for chimerism. Here, for the first time, we reveal advanced pluripotent stem cells (ASCs), which are developmentally beyond the pluripotent cells in the inner cell mass but with higher potency than EpiSCs. Accordingly, a single ASC contributes very efficiently to the fetus, germline, yolk sac and the placental labyrinth in chimeras. Since they are developmentally more advanced, ASCs do not contribute to the trophoblast. ASCs were derived from blastocysts in two steps in a chemically defined medium supplemented with Activin A and basic fibroblast growth factor, followed by culturing in ABCL medium containing ActA, BMP4, CHIR99021 and leukemia inhibitory factor. Notably, ASCs exhibit a distinct transcriptome with the expression of both naive pluripotency genes, as well as mesodermal somatic genes; Eomes, Eras, Tdgf1, Evx1, hand1, Wnt5a and distinct repetitive elements. Conversion of established ESCs to ASCs is also achievable. Importantly, ASCs exhibit a stable hypermethylated epigenome and mostly intact imprints as compared to the hypomethylated inner cell mass of blastocysts and naive ESCs. Properties of ASCs suggest that they represent cells at an intermediate cellular state between the naive and primed states of pluripotency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Chimera
  • DNA Methylation
  • Epidermal Growth Factor / genetics
  • Germ Layers / cytology
  • Homeodomain Proteins / genetics
  • Membrane Glycoproteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mouse Embryonic Stem Cells* / cytology
  • Mouse Embryonic Stem Cells* / metabolism
  • Neoplasm Proteins / genetics
  • Oncogene Protein p21(ras) / genetics
  • Sequence Analysis, RNA
  • T-Box Domain Proteins / genetics
  • Wnt-5a Protein / genetics

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • ERas protein, mouse
  • Eomes protein, mouse
  • Evx1 protein, mouse
  • Hand1 protein, mouse
  • Homeodomain Proteins
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • T-Box Domain Proteins
  • Tdgf1 protein, mouse
  • Wnt-5a Protein
  • Wnt5a protein, mouse
  • Epidermal Growth Factor
  • Oncogene Protein p21(ras)