Multipotent RAG1+ progenitors emerge directly from haemogenic endothelium in human pluripotent stem cell-derived haematopoietic organoids.
Ali Motazedian, Freya F Bruveris, Santhosh V Kumar, Jacqueline V Schiesser, Tyrone Chen, Elizabeth S Ng, Ann P Chidgey, Christine A Wells, Andrew G Elefanty, Edouard G Stanley
Author Information
- Ali Motazedian: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Freya F Bruveris: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Santhosh V Kumar: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Jacqueline V Schiesser: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Tyrone Chen: Centre for Stem Cell Systems, Anatomy and Neuroscience, University of Melbourne, Parkville, Victoria, Australia.
- Elizabeth S Ng: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Ann P Chidgey: Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
- Christine A Wells: Centre for Stem Cell Systems, Anatomy and Neuroscience, University of Melbourne, Parkville, Victoria, Australia. ORCID
- Andrew G Elefanty: Murdoch Children's Research Institute, Parkville, Victoria, Australia.
- Edouard G Stanley: Murdoch Children's Research Institute, Parkville, Victoria, Australia. ed.stanley@mcri.edu.au. ORCID
Defining the ontogeny of the human adaptive immune system during embryogenesis has implications for understanding childhood diseases including leukaemias and autoimmune conditions. Using RAG1:GFP human pluripotent stem cell reporter lines, we examined human T-cell genesis from pluripotent-stem-cell-derived haematopoietic organoids. Under conditions favouring T-cell development, RAG1+ cells progressively upregulated a cohort of recognized T-cell-associated genes, arresting development at the CD4+CD8+ stage. Sort and re-culture experiments showed that early RAG1+ cells also possessed B-cell, myeloid and erythroid potential. Flow cytometry and single-cell-RNA-sequencing data showed that early RAG1+ cells co-expressed the endothelial/haematopoietic progenitor markers CD34, VECAD and CD90, whereas imaging studies identified RAG1+ cells within CD31+ endothelial structures that co-expressed SOX17+ or the endothelial marker CAV1. Collectively, these observations provide evidence for a wave of human T-cell development that originates directly from haemogenic endothelium via a RAG1+ intermediate with multilineage potential.
Cell Differentiation
Cell Line
Embryonic Development
Endothelium
Hemangioblasts
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells
Homeodomain Proteins
Humans
Organoids
Pluripotent Stem Cells
Homeodomain Proteins
RAG-1 protein
