Human definitive haemogenic endothelium and arterial vascular endothelium represent distinct lineages

Andrea Ditadi; Christopher M Sturgeon; Joanna Tober; Geneve Awong; Marion Kennedy; Amanda D Yzaguirre; Lisa Azzola; Elizabeth S Ng; Edouard G Stanley; Deborah L French; Xin Cheng; Paul Gadue; Nancy A Speck; Andrew G Elefanty; Gordon Keller

doi:10.1038/ncb3161

Human definitive haemogenic endothelium and arterial vascular endothelium represent distinct lineages

Nat Cell Biol. 2015 May;17(5):580-91. doi: 10.1038/ncb3161. Epub 2015 Apr 27.

Authors

Affiliations

¹ McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
² Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
³ Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria 3052, Australia.
⁴ 1] Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria 3052, Australia [2] Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University Clayton, Victoria 3052, Australia.
⁵ 1] Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria 3052, Australia [2] Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University Clayton, Victoria 3052, Australia [3] Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3052, Australia.
⁶ Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.

PMID: 25915127
PMCID: PMC4551438
DOI: 10.1038/ncb3161

Abstract

The generation of haematopoietic stem cells (HSCs) from human pluripotent stem cells (hPSCs) will depend on the accurate recapitulation of embryonic haematopoiesis. In the early embryo, HSCs develop from the haemogenic endothelium (HE) and are specified in a Notch-dependent manner through a process named endothelial-to-haematopoietic transition (EHT). As HE is associated with arteries, it is assumed that it represents a subpopulation of arterial vascular endothelium (VE). Here we demonstrate at a clonal level that hPSC-derived HE and VE represent separate lineages. HE is restricted to the CD34(+)CD73(-)CD184(-) fraction of day 8 embryoid bodies and it undergoes a NOTCH-dependent EHT to generate RUNX1C(+) cells with multilineage potential. Arterial and venous VE progenitors, in contrast, segregate to the CD34(+)CD73(med)CD184(+) and CD34(+)CD73(hi)CD184(-) fractions, respectively. Together, these findings identify HE as distinct from VE and provide a platform for defining the signalling pathways that regulate their specification to functional HSCs.

Publication types

Comparative Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Video-Audio Media

MeSH terms

5'-Nucleotidase / deficiency
Antigens, CD34 / metabolism
Arteries / cytology
Arteries / metabolism
Arteries / physiology*
Biomarkers / metabolism
Cell Differentiation*
Cell Line
Cell Lineage*
Cell Separation / methods
Coculture Techniques
Core Binding Factor Alpha 2 Subunit / metabolism
Endothelial Progenitor Cells / metabolism
Endothelial Progenitor Cells / physiology*
GPI-Linked Proteins / deficiency
Hematopoietic Stem Cells / metabolism
Hematopoietic Stem Cells / physiology*
Humans
Microscopy, Video
Multipotent Stem Cells / metabolism
Multipotent Stem Cells / physiology*
Phenotype
Pluripotent Stem Cells / metabolism
Pluripotent Stem Cells / physiology*
Precursor Cells, T-Lymphoid / physiology
Receptors, CXCR5 / deficiency
Receptors, Notch / metabolism
Signal Transduction
Time Factors
Veins / cytology
Veins / physiology

Substances

Antigens, CD34
Biomarkers
Core Binding Factor Alpha 2 Subunit
GPI-Linked Proteins
RUNX1 protein, human
Receptors, CXCR5
Receptors, Notch
5'-Nucleotidase
NT5E protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding