Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development.
Jonathan HenningerBuyung SantosoStefan HansEllen DurandJessica MooreChristian MosimannMichael BrandDavid TraverLeonard ZonPublished in: Nature cell biology (2016)
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.
Keyphrases
- stem cells
- single cell
- cell therapy
- aortic valve
- rna seq
- high resolution
- genome wide
- pulmonary artery
- high throughput
- gene expression
- left ventricular
- cancer therapy
- pulmonary arterial hypertension
- aortic dissection
- radiation therapy
- dna methylation
- low birth weight
- preterm infants
- radiation induced
- mesenchymal stem cells