Human definitive haemogenic endothelium and arterial vascular endothelium represent distinct lineages.
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.
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Andrea
A
Ditadi
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Christopher M
CM
Sturgeon
Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Joanna
J
Tober
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Geneve
G
Awong
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Marion
M
Kennedy
Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Amanda D
AD
Yzaguirre
Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria 3052, Australia.
Lisa
L
Azzola
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.
Elizabeth S
ES
Ng
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.
Edouard G
EG
Stanley
Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
Deborah L
DL
French
Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
Xin
X
Cheng
Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
Paul
P
Gadue
Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Nancy A
NA
Speck
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.
Andrew G
AG
Elefanty
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario M5G 1L7, Canada.
Gordon
G
Keller
D015720:5'-Nucleotidase / Q000172:deficiency; D018952:Antigens, CD34 / Q000378:metabolism; D001158:Arteries / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism; D066026:Endothelial Progenitor Cells / Q000378:metabolism / Q000502:physiology*; D058851:GPI-Linked Proteins / Q000172:deficiency; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D018715:Microscopy, Video; D039902:Multipotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D054504:Precursor Cells, T-Lymphoid / Q000502:physiology; D054380:Receptors, CXCR5 / Q000172:deficiency; D051880:Receptors, Notch / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014680:Veins / Q000166:cytology / Q000502:physiology
D018952:Antigens, CD34; D015415:Biomarkers; D050676:Core Binding Factor Alpha 2 Subunit; D058851:GPI-Linked Proteins; C493728:RUNX1 protein, human; D054380:Receptors, CXCR5; D051880:Receptors, Notch; D015720:5'-Nucleotidase; C548241:NT5E protein, human