Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development
Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development
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DOI：
10.1038/ncb3444
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发表时间：
2017-01-01
影响因子：
21.3
通讯作者：
Zon, Leonard
中科院分区：
文献类型：
--
作者：
Henninger, Jonathan;Santoso, Buyung;Zon, Leonard
造血干细胞（HSCs）在胚胎发生期间在发育中的主动脉中产生。已经通过移植估计了出生的HSC克隆的数量，但是评估在天然环境中形成的HSC的绝对数量的实验方法仍然具有挑战性。在这里，我们应用斑马鱼HSC的单细胞和克隆分析来量化发育中的HSC。在开发cd41：eGFP（+）HSC中靶向creER（T2）使得能够长期评估其血液贡献。我们还应用了基于Brainbow的多色Zebrabow系统，其具有在早期造血中活跃的drhcreER（T2），以诱导每个HSC及其后代独特的可遗传的颜色条形码。我们的研究结果表明，大约21个HSC克隆存在之前，HSC的出现和30个克隆存在于生产高峰期从主动脉内皮。我们的方法进一步揭示了应激造血，包括亚致死辐射和移植，减少克隆多样性。我们的研究结果提供了定量的见解早期克隆事件，调节造血发育。
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.