Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development

Nat Cell Biol. 2017 Jan;19(1):17-27. doi: 10.1038/ncb3444. Epub 2016 Nov 21.


Haematopoietic stem cells (HSCs) arise in the developing aorta during embryogenesis. The number of HSC clones born has been estimated through transplantation, but experimental approaches to assess the absolute number of forming HSCs in a native setting have remained challenging. Here, we applied single-cell and clonal analysis of HSCs in zebrafish to quantify developing HSCs. Targeting creERT2 in developing cd41:eGFP+ HSCs enabled long-term assessment of their blood contribution. We also applied the Brainbow-based multicolour Zebrabow system with drl:creERT2 that is active in early haematopoiesis to induce heritable colour barcoding unique to each HSC and its progeny. Our findings reveal that approximately 21 HSC clones exist prior to HSC emergence and 30 clones are present during peak production from aortic endothelium. Our methods further reveal that stress haematopoiesis, including sublethal irradiation and transplantation, reduces clonal diversity. Our findings provide quantitative insights into the early clonal events that regulate haematopoietic development.

MeSH terms

  • Aging
  • Animals
  • Blood Vessels / embryology
  • Bone Marrow Cells / cytology
  • Bone Marrow Transplantation
  • Cell Count
  • Cell Lineage*
  • Clone Cells
  • Embryo, Nonmammalian / cytology
  • Embryonic Development*
  • Erythroid Cells / cytology
  • Granulocytes / cytology
  • Hematopoiesis
  • Hematopoietic Stem Cells / cytology*
  • Lasers
  • Platelet Membrane Glycoprotein IIb / metabolism
  • Staining and Labeling
  • Transgenes
  • Zebrafish / embryology


  • Platelet Membrane Glycoprotein IIb