title: Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development.
abstract: 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 creERT2in developing cd41:eGFP+HSCs enabled long-term assessment of their blood contribution. We also applied the Brainbow-based multicolour Zebrabow system with drl:creERT2that 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.
authors.affiliation: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.|Biological and Biomedical Sciences, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Henninger
authors.affiliation: Section of Cell and Developmental Biology, University of California at San Diego, La Jolla, California 92093, USA.
authors.forename: Buyung
authors.initials: B
authors.lastname: Santoso
authors.affiliation: Biotechnology Center and Center for Regenerative Therapies Dresden, Dresden University of Technology, Tatzberg 47-49, 01307 Dresden, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hans
authors.affiliation: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.|Biological and Biomedical Sciences, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Durand
authors.affiliation: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Moore
authors.affiliation: Institute of Molecular Life Sciences, University of Zürich, 8057 Zürich, Switzerland.
authors.forename: Christian
authors.identifier: http://orcid.org/0000-0002-0749-2576
authors.initials: C
authors.lastname: Mosimann
authors.affiliation: Biotechnology Center and Center for Regenerative Therapies Dresden, Dresden University of Technology, Tatzberg 47-49, 01307 Dresden, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Brand
authors.affiliation: Section of Cell and Developmental Biology, University of California at San Diego, La Jolla, California 92093, USA.|Department of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, California 92093-0380, USA.
authors.forename: David
authors.initials: D
authors.lastname: Traver
authors.affiliation: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.|Biological and Biomedical Sciences, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Leonard
authors.initials: L
authors.lastname: Zon
mesh_terms: D000375:Aging; D000818:Animals; D001808:Blood Vessels / Q000196:embryology; D001854:Bone Marrow Cells / Q000166:cytology; D016026:Bone Marrow Transplantation; D002452:Cell Count; D019070:Cell Lineage; D002999:Clone Cells; D004625:Embryo, Nonmammalian / Q000166:cytology; D047108:Embryonic Development; D041905:Erythroid Cells / Q000166:cytology; D006098:Granulocytes / Q000166:cytology; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007834:Lasers; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D013194:Staining and Labeling; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology
chemical_list: D040201:Platelet Membrane Glycoprotein IIb