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Erythropoiesis at a clonal level
tion. It would be of interest to examine megakaryocytes/platelet output in our model, at steady state or under stress, but the size and lack of nucleus in platelets present challenges, as does the rarity of megakaryocytes in the BM. We are developing an opti- mized vector allowing concurrent single cell RNA-Seq and barcode retrieval to further investigate platelet and other lineage relationships.17
In conclusion, this study is the first to quantitatively track erythropoiesis at a clonal level in vivo in a translation- ally-relevant model. Overall, our analyses indicate long- term shared ontogeny with other lineages, particularly myeloid cells, without measurable contributions from highly erythroid-biased or restricted long-term engrafting HSPC, even under stressors such as aging or erythroid lin-
eage-specific cytokine stimulation. A better understanding of erythropoiesis at this level has relevance for further development of new therapies targeting erythroid disor- ders.
Funding
This research was supported by the NHLBI Division of Intramural Research.
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Acknowledgments
We thank Naoya Uchida for the χHIV plasmid, and Keyvan Keyvanfar for cell sorting. We acknowledge the support of the NHLBI FACS Core, the NHLBI DNA Sequencing and Genomics Core, the NHLBI animal care and veterinary staff, and the NIH Biowulf High-Performance Computing Resource.
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