Single-Cell Analysis Identifies Distinct Stages of Human Endothelial-to-Hematopoietic Transition

Cell Rep. 2017 Apr 4;19(1):10-19. doi: 10.1016/j.celrep.2017.03.023.


During development, hematopoietic cells originate from endothelium in a process known as endothelial-to-hematopoietic transition (EHT). To study human EHT, we coupled flow cytometry and single-cell transcriptional analyses of human pluripotent stem cell-derived CD34+ cells. The resulting transcriptional hierarchy showed a continuum of endothelial and hematopoietic signatures. At the interface of these two signatures, a unique group of cells displayed both an endothelial signature and high levels of key hematopoietic stem cell-associated genes. This interphase group was validated via sort and subculture as an immediate precursor to hematopoietic cells. Differential expression analyses further divided this population into subgroups, which, upon subculture, showed distinct hematopoietic lineage differentiation potentials. We therefore propose that immediate precursors to hematopoietic cells already have their hematopoietic lineage restrictions defined prior to complete downregulation of the endothelial signature. These findings increase our understanding of the processes of de novo hematopoietic cell generation in the human developmental context.

Keywords: developmental hematopoiesis; endothelial-to-hematopoietic transition; hematopoietic stem cell; human induced pluripotent stem cells; single cell analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism
  • Cell Lineage
  • Cells, Cultured
  • Down-Regulation
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Endothelium / metabolism
  • Flow Cytometry
  • Hematopoiesis / physiology*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • Leukosialin / metabolism
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-ets / metabolism
  • Repressor Proteins / metabolism
  • Single-Cell Analysis
  • Wiskott-Aldrich Syndrome Protein / metabolism


  • Antigens, CD34
  • ETS translocation variant 6 protein
  • GFI1B protein, human
  • Leukosialin
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-ets
  • Repressor Proteins
  • WAS protein, human
  • Wiskott-Aldrich Syndrome Protein