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.

Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution

Nature Medicine volume 16, pages 232–236 (2010)Cite this article

Subjects

Abstract

Delayed myeloid engraftment after cord blood transplantation (CBT) is thought to result from inadequate numbers of progenitor cells in the graft and is associated with increased early transplant–related morbidity and mortality. New culture strategies that increase the number of cord blood progenitors capable of rapid myeloid engraftment after CBT would allow more widespread use of this stem cell source for transplantation. Here we report the development of a clinically relevant Notch-mediated ex vivo expansion system for human CD34+ cord blood progenitors that results in a marked increase in the absolute number of stem/progenitor cells, including those capable of enhanced repopulation in the marrow of immunodeficient nonobese diabetic–severe combined immunodeficient (NOD-SCID) mice. Furthermore, when cord blood progenitors expanded ex vivo in the presence of Notch ligand were infused in a clinical setting after a myeloablative preparative regimen for stem cell transplantation, the time to neutrophil recovery was substantially shortened. To our knowledge, this is the first instance of rapid engraftment derived from ex vivo expanded stem/progenitor cells in humans.

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

Relevant articles

Open Access articles citing this article.

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: Culture of CD34+ cord blood progenitors with Notch ligand results in enhanced and more rapid human engraftment as well as increased SRC frequency.
Figure 2: Clinical-grade culture of cord blood progenitors with Delta1ext-IgG results in marked in vitro expansion of CD34+ cells and more rapid neutrophil recovery in a myeloablative double-CBT setting.

References

  1. Shpall, E.J. et al. Transplantation of ex vivo expanded cord blood. Biol. Blood Marrow Transplant. 8, 368–376 (2002).

    Google Scholar 

  2. de Lima, M. et al. Transplantation of ex vivo expanded cord blood cells using the copper chelator tetraethylenepentamine: a phase I/II clinical trial. Bone Marrow Transplant. 41, 771–778 (2008).

    Google Scholar 

  3. Jaroscak, J. et al. Augmentation of umbilical cord blood (UCB) transplantation with ex vivo–expanded UCB cells: results of a phase 1 trial using the AastromReplicell System. Blood 101, 5061–5067 (2003).

    Google Scholar 

  4. Milner, L.A., Kopan, R., Martin, D.I. & Bernstein, I.D. A human homologue of the Drosophila developmental gene, Notch, is expressed in CD34+ hematopoietic precursors. Blood 83, 2057–2062 (1994).

    Google Scholar 

  5. Varnum-Finney, B. et al. Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling. Nat. Med. 6, 1278–1281 (2000).

    Google Scholar 

  6. Varnum-Finney, B., Brashem-Stein, C. & Bernstein, I.D. Combined effects of Notch signaling and cytokines induce a multiple log increase in precursors with lymphoid and myeloid reconstituting ability. Blood 101, 1784–1789 (2003).

    Google Scholar 

  7. Delaney, C., Varnum-Finney, B., Aoyama, K., Brashem-Stein, C. & Bernstein, I.D. Dose-dependent effects of the Notch ligand Delta1 on ex vivo differentiation and in vivo marrow repopulating ability of cord blood cells. Blood 106, 2693–2699 (2005).

    Google Scholar 

  8. Ohishi, K., Varnum-Finney, B. & Bernstein, I.D. Delta-1 enhances marrow and thymus repopulating ability of human CD34+CD38− cord blood cells. J. Clin. Invest. 110, 1165–1174 (2002).

    Google Scholar 

  9. Bhatia, M. et al. Quantitative analysis reveals expansion of human hematopoietic repopulating cells after short-term ex vivo culture. J. Exp. Med. 186, 619–624 (1997).

    Google Scholar 

  10. Conneally, E., Cashman, J., Petzer, A. & Eaves, C. Expansion in vitro of transplantable human cord blood stem cells demonstrated using a quantitative assay of their lympho-myeloid repopulating activity in nonobese diabetic-scid/scid mice. Proc. Natl. Acad. Sci. USA 94, 9836–9841 (1997).

    Google Scholar 

  11. Suzuki, T. et al. Highly efficient ex vivo expansion of human hematopoietic stem cells using Delta1-Fc chimeric protein. Stem Cells 24, 2456–2465 (2006).

    Google Scholar 

  12. Offner, F., Schoch, G., Fisher, L.D., Torok-Storb, B. & Martin, P.J. Mortality hazard functions as related to neutropenia at different times after marrow transplantation. Blood 88, 4058–4062 (1996).

    Google Scholar 

  13. Nitsche, A. et al. Inhibition of cord blood cell expansion by human herpesvirus 6 in vitro. Stem Cells Dev. 13, 197–203 (2004).

    Google Scholar 

  14. Barker, J.N. et al. Transplantation of 2 partially HLA-matched umbilical cord blood units to enhance engraftment in adults with hematologic malignancy. Blood 105, 1343–1347 (2005).

    Google Scholar 

  15. Bradstock, K. et al. Single versus double unrelated umbilical cord blood units for allogeneic transplantation in adults with advanced hematological malignancies: a retrospective comparison of outcomes. Intern. Med. J. 39, 744–751 (2009).

    Google Scholar 

  16. Brunstein, C.G. et al. Umbilical cord blood transplantation after nonmyeloablative conditioning: impact on transplantation outcomes in 110 adults with hematologic disease. Blood 110, 3064–3070 (2007).

    Google Scholar 

  17. Kang, H.J. et al. Early engraftment kinetics of two units cord blood transplantation. Bone Marrow Transplant. 38, 197–201 (2006).

    Google Scholar 

  18. Rubinstein, P. et al. Processing and cryopreservation of placental/umbilical cord blood for unrelated bone marrow reconstitution. Proc. Natl. Acad. Sci. USA 92, 10119–10122 (1995).

    Google Scholar 

  19. Peled, A. et al. Dependence of human stem cell engraftment and repopulation of NOD/SCID mice on CXCR4. Science 283, 845–848 (1999).

    Google Scholar 

  20. Gutman, J.A. et al. Single unit dominance following double unit umbilical cord blood transplantation coincides with a specific cd8+ t cell response against the non-engrafted unit. Blood published online, doi:10.1182/blood-2009-07-228999 (12 October 2009).

Download references

Acknowledgements

This work was supported by grants R24 HL74445, K23 HL077446, K12 CA076930, 5R01HL080245-02 (from the National Institutes of Health), CA15704 and DK56465. C.D. is a Damon Runyon Clinical Investigator supported in part by the Damon Runyon Cancer Research Foundation (CI# 35-07). I.D.B. is an American Cancer Society–F.M. Kirby Clinical Research Professor. We would like to thank F. Appelbaum for his contribution to clinical trial design and clinical consultation, J. Gutman for help in manuscript editing and clinical data coordination and D. Woodle for expert technical assistance. We also acknowledge the work of the clinical faculty and staff at the Seattle Cancer Care Alliance for the excellent clinical care provided to the patients.

Author information

Authors and Affiliations

  1. Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA

    Colleen Delaney, Shelly Heimfeld, Carolyn Brashem-Stein, Howard Voorhies, Ronald L Manger & Irwin D Bernstein

  2. Department of Pediatrics, University of Washington, Seattle, Washington, USA

    Colleen Delaney & Irwin D Bernstein

Authors
  1. Colleen Delaney
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shelly Heimfeld
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carolyn Brashem-Stein
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Howard Voorhies
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ronald L Manger
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Irwin D Bernstein
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

C.D. conceived of the project, designed the experiments, analyzed the data, wrote the manuscript, performed in vitro and mouse studies, was principal investigator of clinical studies and prepared the figures; S.H. contributed to experimental design, data analysis and manuscript editing; C.B.-S. and H.V. performed in vitro studies, data analysis and mouse studies; R.L.M. contributed methods for large-scale production of cGMP-engineered Delta1; I.D.B. conceived of the project and contributed to experimental design and manuscript editing.

Corresponding author

Correspondence to Irwin D Bernstein.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1 and 2, Supplementary Tables 1–3, Supplementary Methods (PDF 173 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Delaney, C., Heimfeld, S., Brashem-Stein, C. et al. Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med 16, 232–236 (2010). https://doi.org/10.1038/nm.2080

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2080

This article is cited by

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