2. Academic Validation
  3. Protocol for the Generation of Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

Protocol for the Generation of Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

  • STAR Protoc. 2020 Oct 16;1(3):100130. doi: 10.1016/j.xpro.2020.100130.
Monica Nafria 1 2 3,  Constanze Bonifer 1,  Edouard Guy Stanley 2 3 4,  Elizabeth Siewsun Ng 2,  Andrew George Elefanty 2 3 4
Affiliations

Affiliations

  • 1 Institute for Cancer and Genomic Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham B152TT, UK.
  • 2 Murdoch Children's Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, VIC 3052, Australia.
  • 3 Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC 3052, Australia.
  • 4 Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
PMID: 33377024 DOI: 10.1016/j.xpro.2020.100130
Abstract

This protocol offers a detailed procedure for the in vitro differentiation of human pluripotent stem cells (hPSCs) to multipotent hematopoietic progenitors that arise from SOX17+ hemogenic endothelium, mimicking intra-embryonic, HOXA-positive, aorta-gonad mesonephros (AGM) hematopoiesis. The generated endothelium displays transcriptional similarities to cells sorted from human 5-week AGM, and CD45+CD34+RUNX1C+ progenitors share an accessible chromatin profile with adult hematopoietic stem cells and multipotent progenitors. Therefore, this protocol is suitable for the mechanistic study of human multipotent progenitor development and for modeling childhood leukemias. For complete details on the use and execution of this protocol, please refer to Nafria et al. (2020).

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