title: Single-Cell Analysis Identifies Distinct Stages of Human Endothelial-to-Hematopoietic Transition.
abstract: 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.
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden.
authors.forename: Carolina
authors.initials: C
authors.lastname: Guibentif
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden.
authors.forename: Roger Emanuel
authors.initials: RE
authors.lastname: Rönn
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Böiers
authors.affiliation: Division of Molecular Hematology, Lund Stem Cell Center, Lund University, BMC B12, 221 84 Lund, Sweden.
authors.forename: Stefan
authors.initials: S
authors.lastname: Lang
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden.
authors.forename: Shobhit
authors.initials: S
authors.lastname: Saxena
authors.affiliation: Division of Molecular Hematology, Lund Stem Cell Center, Lund University, BMC B12, 221 84 Lund, Sweden.
authors.forename: Shamit
authors.initials: S
authors.lastname: Soneji
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden; Stem Cell Laboratory, UCL Cancer Institute, University College London, London W1CE 6BT, UK.
authors.forename: Tariq
authors.initials: T
authors.lastname: Enver
authors.affiliation: Division of Molecular Hematology, Lund Stem Cell Center, Lund University, BMC B12, 221 84 Lund, Sweden. Electronic address: goran.karlsson@med.lu.se.
authors.forename: Göran
authors.initials: G
authors.lastname: Karlsson
authors.affiliation: Section of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden. Electronic address: niels-bjarne.woods@med.lu.se.
authors.forename: Niels-Bjarne
authors.initials: NB
authors.lastname: Woods
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D019070:Cell Lineage; D002478:Cells, Cultured; D015536:Down-Regulation; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004727:Endothelium / Q000378:metabolism; D005434:Flow Cytometry; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051917:Leukosialin / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D059010:Single-Cell Analysis; D051316:Wiskott-Aldrich Syndrome Protein / Q000378:metabolism; D000097764:ETS Translocation Variant 6 Protein
chemical_list: D018952:Antigens, CD34; D051917:Leukosialin; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012097:Repressor Proteins; D051316:Wiskott-Aldrich Syndrome Protein; D000097764:ETS Translocation Variant 6 Protein; C111977:GFI1B protein, human; C495767:WAS protein, human
keywords: developmental hematopoiesis; endothelial-to-hematopoietic transition; hematopoietic stem cell; human induced pluripotent stem cells; single cell analysis