Isolation of primitive endoderm, mesoderm, vascular endothelial and trophoblast progenitors from human pluripotent stem cells

Nat Biotechnol. 2012 May 27;30(6):531-42. doi: 10.1038/nbt.2239.

Abstract

To identify early populations of committed progenitors derived from human embryonic stem cells (hESCs), we screened self-renewing, BMP4-treated and retinoic acid-treated cultures with >400 antibodies recognizing cell-surface antigens. Sorting of >30 subpopulations followed by transcriptional analysis of developmental genes identified four distinct candidate progenitor groups. Subsets detected in self-renewing cultures, including CXCR4(+) cells, expressed primitive endoderm genes. Expression of Cxcr4 in primitive endoderm was confirmed in visceral endoderm of mouse embryos. BMP4-induced progenitors exhibited gene signatures of mesoderm, trophoblast and vascular endothelium, suggesting correspondence to gastrulation-stage primitive streak, chorion and allantois precursors, respectively. Functional studies in vitro and in vivo confirmed that ROR2(+) cells produce mesoderm progeny, APA(+) cells generate syncytiotrophoblasts and CD87(+) cells give rise to vasculature. The same progenitor classes emerged during the differentiation of human induced pluripotent stem cells (hiPSCs). These markers and progenitors provide tools for purifying human tissue-regenerating progenitors and for studying the commitment of pluripotent stem cells to lineage progenitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Surface / chemistry
  • Biomarkers / chemistry
  • Cluster Analysis
  • Endoderm / chemistry
  • Endoderm / cytology*
  • Endothelium, Vascular / chemistry
  • Endothelium, Vascular / cytology*
  • Flow Cytometry
  • Glutamyl Aminopeptidase / genetics
  • Humans
  • Mesoderm / chemistry
  • Mesoderm / cytology*
  • Mice
  • Mice, Transgenic
  • Pluripotent Stem Cells / chemistry
  • Pluripotent Stem Cells / classification
  • Pluripotent Stem Cells / cytology*
  • Receptors, CXCR4 / biosynthesis
  • Receptors, CXCR4 / genetics
  • Trophoblasts / chemistry
  • Trophoblasts / cytology*

Substances

  • Antigens, Surface
  • Biomarkers
  • Receptors, CXCR4
  • ENPEP protein, human
  • Glutamyl Aminopeptidase