Analysis of histone 2B-GFP retention reveals slowly cycling hematopoietic stem cells
Open accessAbstract
Hematopoietic stem cells (HSCs) are thought to divide infrequently based on their resistance to cytotoxic injury targeted at rapidly cycling cells and have been presumed to retain labels such as the thymidine analog 5-bromodeoxyuridine (BrdU). However, BrdU retention is neither a sensitive nor specific marker for HSCs. Here we show that transient, transgenic expression of a histone 2B (H2B)-green fluorescent protein (GFP) fusion protein in mice has several advantages for label-retention studies over BrdU, including rapid induction of H2B-GFP in virtually all HSCs, higher labeling intensity and the ability to prospectively study label-retaining cells, which together permit a more precise analysis of division history. Mathematical modeling of H2B-GFP dilution in HSCs, identified with a stringent marker combination (L(-)K(+)S(+)CD48(-)CD150(+)), revealed unexpected heterogeneity in their proliferation rates and showed that approximately 20% of HSCs divide at an extremely low rate (< or =0.8-1.8% per day).
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Slow cycling is also observed with HSCs, and long-term repopulating potential is inversely correlated with cycling.
“Similarly, slow cycling is also observed with HSCs, and long-term repopulating potential is inversely correlated with cycling [47].”
From Clinical use of lentiviral vectors (2018) · cites this paper as [47]
The paper reports a slowly cycling HSC-enriched population and finds that greater H2B-GFP retention correlates with better long-term repopulation after transplantation (passages [3] and [6], indexed under “Abstract”). But it explicitly cautions that it cannot unambiguously conclude that HSC proliferation history predicts repopulation potential, because the marker-defined population may contain both stem and progenitor cells (passage [1]). The claim omits that qualification. (LLM, conf=0.84)
Experimental — accuracy not yet measuredLoss of quiescence is a major factor leading to erosion of hematopoietic stem cell self-renewal.
“Loss of quiescence is a major factor leading to erosion of HSC self-renewal (31,32).”
From <i>Txnip</i> Enhances Fitness of <i>Dnmt3a</i> -Mutant Hematopoietic Stem Cells via <i>p21</i> (2022) · cites this paper as [32]
The passage labeled Abstract (page unavailable) reports that greater H2B-GFP retention correlates with stronger long-term repopulation potential, consistent with a link between slower cycling and HSC function. But the authors explicitly say they cannot establish that proliferation history predicts repopulation potential; they present division-driven loss of self-renewal as speculation, not as an established major cause. (LLM, conf=0.87)
Experimental — accuracy not yet measuredBecause many HSCs remain quiescent for much of adult life, continuing DNA damage accumulation may eventually exceed a threshold that impairs stem cell function in aged animals.
“Thus, as many HSCs remain quiescent for much of adult life54,55, the continuing accumulation of damage in these cells may eventually exceed a threshold that impairs stem cell function in aged animals.”
From Stem cell aging: mechanisms, regulators and therapeutic opportunities (2014) · cites this paper as [55]
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- Authors
- Foudi, A., Hochedlinger, K., Van Buren, D., Schindler, J. W., Jaenisch, R., Carey, V., & Hock, H.
- Affiliations
- Harvard University (US)
- Massachusetts General Hospital (US)
- Whitehead Institute for Biomedical Research (US)
- Brigham and Women's Hospital (US)
- Corresponding author
- Hock, Hanno
- Venue
- Nature Biotechnology
- Published
- 2008-12-05
- Volume
- 27
- Issue
- 1
- Pages
- 84-90
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC2805441
- OpenAlex
- W1968034850
- Cited by
- 586
- References
- 37
- Citations by year
- 2026: 12 · 2025: 21 · 2024: 19 · 2023: 23 · 2022: 34 · 2021: 40 · 2020: 34 · 2019: 39 · 2018: 33 · 2017: 42 · 2016: 46 · 2015: 36 · 2014: 43 · 2013: 40 · 2012: 41
- FWCI
- 18.03
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Cancer Institute (R01CA122726)
- Topics
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Blood disorders and treatments
- Keywords
- hematopoietic stem cells, label retention, cell cycle, BrdU labeling, mathematical modeling
- MeSH terms
- Signaling Lymphocytic Activation Molecule Family Member 1, CD48 Antigen, Animals, Bone Marrow Cells, Bromodeoxyuridine, Cell Differentiation, Flow Cytometry, Hematopoietic Stem Cells, Histones, Humans, Models, Theoretical, Receptors, Cell Surface, Antigens, CD, Hematopoietic Stem Cell Transplantation, Cell Lineage, Green Fluorescent Proteins, Mice
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