Poster Presentation ASSCR, AGCTS, ISCT ANZ and Friends Joint Scientific Conference 2019

Multipotent RAG1+ progenitors emerge directly from hemogenic endothelium in human PSC derived hematopoietic organoids. (#111)

Ali Motazedian 1 , Freya Faith Bruveris 1 , Santhosh V Kumar 1 , Jacqueline V Schiesser 1 , Tyrone Chen 2 , Elizabeth Ng 1 , Ann Chidgey 3 , Christine Wells 2 , Andrew Elefanty 1 , Ed Stanley 1
  1. Murdoch Children's Research Institute, Parkville, VIC, Australia
  2. Centre for Stem Cell Systems, Anatomy and Neuroscience, University of Melbourne, Melbourne, VIC, Australia
  3. Anatomy and Developmental Biology, Monash University, Melbourne, VIC, Australia

Defining the ontogeny of the human adaptive immune system during embryogenesis has implications for understanding childhood diseases including leukemias and autoimmune conditions. Using RAG1:GFP human pluripotent stem cell (PSC) reporter lines, we examined human T-cell genesis from PSC derived hematopoietic organoids. Under conditions favouring T-cell development, RAG1+ cells progressively up regulated a cohort of recognised 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, with B-cell potential correlating with co-expression of the progenitor marker VECAD. Consistent with this, RNAseq analysis of the early RAG1+ cells indicated that, in addition to T-lineage genes, this population expressed genes associated with erythroid and myeloid lineages. In addition, flow cytometry and single cell RNA sequencing data showed that early RAG1+ cells co-expressed the endothelial/hematopoietic progenitor markers CD34, VECAD and CD90 whilst 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 multi-lineage potential.

  • Have you presented your abstract at another international meeting?: Yes