Journal article

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



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 R..

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Awarded by National Health and Medical Research Council of Australia

Funding Acknowledgements

We would like to thank the members of the MCRI Flow cytometry support team for their assistance with cell sorting and microscopy and S. Chen for her assistance with the preparation of the art work. This study was funded by the National Health and Medical Research Council of Australia through research fellowships awarded to A.G.E. (grant no. GNT1117596) and E.G.S (grant no. GNT1079004), project grants awarded to A.G.E. and E.G.S. (grant nos GNT1068866, GNT1129861 and GNT1138717) as well as A.P.C. and E.G.S (grant no. GNT1123277), the Australian Research Council Special Research Initiative in Stem Cells (Stem Cells Australia), The Children's Cancer Research Foundation (Australia) and the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program.