Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Haematopoietic stem cells do not asymmetrically segregate chromosomes or retain BrdU

Nature volume 449, pages 238–242 (2007)Cite this article

Abstract

Stem cells are proposed to segregate chromosomes asymmetrically during self-renewing divisions so that older (‘immortal’) DNA strands are retained in daughter stem cells whereas newly synthesized strands segregate to differentiating cells1,2,3,4,5,6. Stem cells are also proposed to retain DNA labels, such as 5-bromo-2-deoxyuridine (BrdU), either because they segregate chromosomes asymmetrically or because they divide slowly5,7,8,9. However, the purity of stem cells among BrdU-label-retaining cells has not been documented in any tissue, and the ‘immortal strand hypothesis’ has not been tested in a system with definitive stem cell markers. Here we tested these hypotheses in haematopoietic stem cells (HSCs), which can be highly purified using well characterized markers. We administered BrdU to newborn mice, mice treated with cyclophosphamide and granulocyte colony-stimulating factor, and normal adult mice for 4 to 10 days, followed by 70 days without BrdU. In each case, less than 6% of HSCs retained BrdU and less than 0.5% of all BrdU-retaining haematopoietic cells were HSCs, revealing that BrdU has poor specificity and poor sensitivity as an HSC marker. Sequential administration of 5-chloro-2-deoxyuridine and 5-iodo-2-deoxyuridine indicated that all HSCs segregate their chromosomes randomly. Division of individual HSCs in culture revealed no asymmetric segregation of the label. Thus, HSCs cannot be identified on the basis of BrdU-label retention and do not retain older DNA strands during division, indicating that these are not general properties of stem cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Contrasting predictions regarding stem cell labelling on the basis of the immortal strand model versus random chromosome segregation.
Figure 2: Six per cent of HSCs stochastically enter the cell cycle each day.
Figure 3: Few HSCs retain BrdU, and most BrdU-retaining bone marrow cells are not HSCs.
Figure 4: HSCs segregate chromosomes randomly in vivo and in vitro.

Similar content being viewed by others

References

  1. Potten, C. S., Hume, W. J., Reid, P. & Cairns, J. The segregation of DNA in epithelial stem cells. Cell 15, 899–906 (1978)

    Article  CAS  Google Scholar 

  2. Cairns, J. Mutation selection and the natural history of cancer. Nature 255, 197–200 (1975)

    Article  ADS  CAS  Google Scholar 

  3. Karpowicz, P. et al. Support for the immortal strand hypothesis: neural stem cells partition DNA asymmetrically in vitro. J. Cell Biol. 170, 721–732 (2005)

    Article  CAS  Google Scholar 

  4. Smith, G. H. Label-retaining epithelial cells in mouse mammary gland divide asymmetrically and retain their template DNA strands. Development 132, 681–687 (2005)

    Article  CAS  Google Scholar 

  5. Shinin, V., Gayraud-Morel, B., Gomes, D. & Tajbakhsh, S. Asymmetric division and cosegregation of template DNA strands in adult muscle satellite cells. Nature Cell Biol. 8, 677–687 (2006)

    Article  CAS  Google Scholar 

  6. Conboy, M. J., Karasov, A. O. & Rando, T. A. High incidence of non-random template strand segregation and asymmetric fate determination in dividing stem cells and their progeny. PLoS Biol. 5, e102 (2007)

    Article  Google Scholar 

  7. Tumbar, T. et al. Defining the epithelial stem cell niche in skin. Science 303, 359–363 (2004)

    Article  ADS  CAS  Google Scholar 

  8. Cotsarelis, G., Sun, T. T. & Lavker, R. M. Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 61, 1329–1337 (1990)

    Article  CAS  Google Scholar 

  9. Zhang, J. et al. Identification of the haematopoietic stem cell niche and control of the niche size. Nature 425, 836–841 (2003)

    Article  ADS  CAS  Google Scholar 

  10. Armakolas, A. & Klar, A. J. Cell type regulates selective segregation of mouse chromosome 7 DNA strands in mitosis. Science 311, 1146–1149 (2006)

    Article  ADS  CAS  Google Scholar 

  11. Arai, F. et al. Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 118, 149–161 (2004)

    Article  CAS  Google Scholar 

  12. Welm, B. E. et al. Sca-1+ cells in the mouse mammary gland represent an enriched progenitor cell population. Dev. Biol. 245, 42–56 (2002)

    Article  CAS  Google Scholar 

  13. Potten, C. S., Owen, G. & Booth, D. Intestinal stem cells protect their genome by selective segregation of template DNA strands. J. Cell Sci. 115, 2381–2388 (2002)

    Article  CAS  Google Scholar 

  14. Urbanek, K. et al. Stem cell niches in the adult mouse heart. Proc. Natl Acad. Sci. USA 103, 9226–9231 (2006)

    Article  ADS  CAS  Google Scholar 

  15. Cheshier, S. H., Morrison, S. J., Liao, X. & Weissman, I. L. In vivo proliferation and cell cycle kinetics of long-term self-renewing hematopoietic stem cells. Proc. Natl Acad. Sci. USA 96, 3120–3125 (1999)

    Article  ADS  CAS  Google Scholar 

  16. Takano, H., Ema, H., Sudo, K. & Nakauchi, H. Asymmetric division and lineage commitment at the level of hematopoietic stem cells: inference from differentiation in daughter cell and granddaughter cell pairs. J. Exp. Med. 199, 295–302 (2004)

    Article  CAS  Google Scholar 

  17. Kiel, M. J., Yilmaz, O. H., Iwashita, T., Terhorst, C. & Morrison, S. J. SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell 121, 1109–1121 (2005)

    Article  CAS  Google Scholar 

  18. Bowie, M. B. et al. Hematopoietic stem cells proliferate until after birth and show a reversible phase-specific engraftment defect. J. Clin. Invest. 116, 2808–2816 (2006)

    Article  CAS  Google Scholar 

  19. Morrison, S. J., Wright, D. & Weissman, I. L. Cyclophosphamide/granulocyte colony-stimulating factor induces hematopoietic stem cells to proliferate prior to mobilization. Proc. Natl Acad. Sci. USA 94, 1908–1913 (1997)

    Article  ADS  CAS  Google Scholar 

  20. Taylor, G., Lehrer, M. S., Jensen, P. J., Sun, T. T. & Lavker, R. M. Involvement of follicular stem cells in forming not only the follicle but also the epidermis. Cell 102, 451–461 (2000)

    Article  CAS  Google Scholar 

  21. Morrison, S. J. et al. Culture in reduced levels of oxygen promotes clonogenic sympathoadrenal differentiation by isolated neural crest stem cells. J. Neurosci. 20, 7370–7376 (2000)

    Article  CAS  Google Scholar 

  22. Bonhoeffer, S., Mohri, H., Ho, D. & Perelson, A. S. Quantification of cell turnover kinetics using 5-bromo-2'-deoxyuridine. J Immunol. 164, 5049–5054 (2000)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Howard Hughes Medical Institute, the National Institute on Aging (NIH), and the US Army Research Laboratory/Office. Flow cytometry was partially supported by the UM-Comprehensive Cancer Centre and the UM-Multipurpose Arthritis Centre. Antibody production was partially supported by the Rheumatic Core Disease Centre. M.J.K. was supported by a University of Michigan Cancer Biology Training Grant. The authors thank D. Adams and M. White for flow cytometry and E. Smith (Hybridoma Core Facility) for antibody production.

Author Contributions M.J.K. performed all experiments and interpreted results. S.H. assisted in the design and interpretation of many experiments and helped to perform some experiments. R.A., S.N.G. and T.L.J. generated the mathematical model of BrdU retention over time (Fig. 3a). M.T. and J.A.K. developed the protocol for double-labelling cells with CldU and IdU. S.J.M. participated in the design and interpretation of experiments, and wrote the paper with M.J.K. and S.H.

Author information

Authors and Affiliations

  1. Department of Internal Medicine, Howard Hughes Medical Institute, Life Sciences Institute, and Centre for Stem Cell Biology,

    Mark J. Kiel, Shenghui He & Sean J. Morrison

  2. Department of Mathematics, University of Michigan, Ann Arbor, Michigan 48109-2216, USA,

    Rina Ashkenazi, Sara N. Gentry & Trachette L. Jackson

  3. Division of Endocrinology, Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA,

    Monica Teta & Jake A. Kushner

Authors
  1. Mark J. Kiel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shenghui He
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rina Ashkenazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sara N. Gentry
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Monica Teta
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jake A. Kushner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Trachette L. Jackson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sean J. Morrison
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sean J. Morrison.

Ethics declarations

Competing interests

Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures S1-S5 with Legends. (PDF 1462 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kiel, M., He, S., Ashkenazi, R. et al. Haematopoietic stem cells do not asymmetrically segregate chromosomes or retain BrdU. Nature 449, 238–242 (2007). https://doi.org/10.1038/nature06115

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature06115

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing