期刊
NATURE CELL BIOLOGY
卷 19, 期 1, 页码 17-27出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/ncb3444
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资金
- HHMI
- NIH [F31HL126338, R01-DK074482, F32DK752433, R01-HL04880, P01-HL032262, 5P30-DK49216, 5R01-DK53298, 5U01-HL10001, R24-DK09276]
- Deutsche Forschungsgemeinschaft [SFB655]
- TU Dresden
- European Union (Zf Health)
- MPN Research Foundation Grant
- EMBO long-term fellowship
- HSFP long-term fellowship
- SNF advanced fellowship
- SNF professorship
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 creER(T2) in developing cd41:eGFP(+) HSCs enabled long-term assessment of their blood contribution. We also applied the Brainbow-based multicolour Zebrabow system with drhcreER(T2) 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.
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