{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T16:02:21Z","timestamp":1771084941730,"version":"3.50.1"},"posted":{"date-parts":[[2026,2,12]]},"group-title":"Cell Biology","reference-count":78,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,12]],"date-time":"2026-02-12T00:00:00Z","timestamp":1770854400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH","award":["R35HL150826"],"award-info":[{"award-number":["R35HL150826"]}]},{"name":"CIRM","award":["EDUC4-12756"],"award-info":[{"award-number":["EDUC4-12756"]}]},{"name":"Leukemia & Lymphoma Society Scholar","award":["LLS-1370-20"],"award-info":[{"award-number":["LLS-1370-20"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Blood and immune cell regeneration is sustained by hematopoietic stem and progenitor cells (HSPCs), which form the therapeutic basis of bone marrow transplantation. While the functional hierarchy of mouse HSPC subsets is well characterized, the distinct roles of human HSPC populations remain less well defined, particularly at clonal resolution and in the context of transplantation conditioning. While clonal tracking in humans and non-human primates has significantly advanced our understanding of hematopoietic dynamics, prior studies predominantly focused on CD34\n                  <jats:sup>+<\/jats:sup>\n                  cells, a heterogeneous population of HSPCs. Moreover, secondary transplantation is considered the gold standard for distinguishing hematopoietic stem cells (HSCs) from multipotent progenitors (MPPs) in mice, but it has not been effectively utilized to study human HSPC populations.\n                <\/jats:p>\n                <jats:p>To address this knowledge gap, we performed quantitative clonal tracking of purified human HSCs (hHSCs) and human MPPs (hMPPs) in NSGW41 mice across primary and secondary transplantation under no conditioning, busulfan, and irradiation. Consistent with prior studies, both hHSCs and hMPPs sustained long-term multilineage reconstitution and differed in engraftment rates. Our quantitative clonal analysis further revealed that hHSC clones generated more blood cells, initiated lymphoid production earlier, and exhibited more robust multilineage differentiation than hMPP clones. hHSC clones were also less sensitive to conditioning, maintaining stable lineage biases. Notably, busulfan and irradiation differentially affected the magnitude, lineage bias, and timing of hematopoietic reconstitution without altering engraftment. During secondary transplantation, hHSCs and hMPPs contributed comparably to hematopoietic reconstitution, but their overall output, particularly monocytes and T cells, was substantially reduced. In contrast to primary recipients, human chimerism of secondary recipients in the peripheral blood was diminished relative to the bone marrow and spleen, and more hHSPC clones contributed to hematopoiesis. Extramedullary hematopoiesis was observed in all secondary recipients, with comparable contributions from hHSC and hMPP clones.<\/jats:p>\n                <jats:p>Overall, this study provides insights into the distinct functions of hHSCs and hMPPs, the influence of conditioning, and the inefficiency of human hematopoiesis through serial transplantation. These findings advance our understanding of human hematopoiesis and provide a framework for utilizing and optimizing experimental models, improving transplantation conditioning strategies, and informing the preclinical evaluation of HSC-based cell and gene therapies.<\/jats:p>","DOI":"10.64898\/2026.02.10.705085","type":"posted-content","created":{"date-parts":[[2026,2,12]],"date-time":"2026-02-12T20:45:18Z","timestamp":1770929118000},"source":"Crossref","is-referenced-by-count":0,"title":["Tracking Human HSCs and MPPs in Mice Reveals Distinct Clonal Dynamics and Responses to Pre-transplant Conditioning"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-9842-5663","authenticated-orcid":false,"given":"Mary","family":"Vergel Snodgrass","sequence":"first","affiliation":[{"name":"Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8940-3786","authenticated-orcid":false,"given":"Jiya","family":"Eerdeng","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patrick","family":"Condie","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5151-9246","authenticated-orcid":false,"given":"Rong","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA"},{"name":"Alfred E. 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