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Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling

Nature Medicine volume 6, pages 1278–1281 (2000)Cite this article

Abstract

Hematopoietic stem cells give rise to progeny that either self-renew in an undifferentiated state or lose self-renewal capabilities and commit to lymphoid or myeloid lineages. Here we evaluated whether hematopoietic stem cell self-renewal is affected by the Notch pathway. Notch signaling controls cell fate choices in both invertebrates and vertebrates1,2,3,4,5,6,7 by inhibiting certain differentiation pathways, thereby permitting cells to either differentiate along an alternative pathway or to self-renew1. Notch receptors are present in hematopoietic precursors and Notch signaling enhances the in vitro generation of human and mouse hematopoietic precursors8,9,10,11,12,13,14,15, determines T- or B-cell lineage specification from a common lymphoid precursor16,17 and promotes expansion of CD8+ cells18,19,20. Here, we demonstrate that constitutive Notch1 signaling in hematopoietic cells established immortalized, cytokine-dependent cell lines that generated progeny with either lymphoid or myeloid characteristics both in vitro and in vivo. These data support a role for Notch signaling in regulating hematopoietic stem cell self-renewal. Furthermore, the establishment of clonal, pluripotent cell lines provides the opportunity to assess mechanisms regulating stem cell commitment and demonstrates a general method for immortalizing stem cell populations for further analysis.

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Figure 1: Retroviral constructs, detection of integration sites and ICN1 expression.
Figure 2: Morphology and phenotype of HSCN1cl10 cells incubated with cytokines.
Figure 3: HSCN1cl10 reconstitutes the bone marrow and thymus with both myeloid and lymphoid cells.

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Acknowledgements

We thank M. Linenberger, S. Collins and K. Ohishi for criticism of the manuscript. This work was supported by grants P50 HL54881, RO1-CA82308-01 and RO1-AI47833-01 from the National Institutes of Health. I.D.B is also supported as an American Cancer Society-F.M. KirbyClinical Research Professorship, and W.S.P is the recipient of a Scholar Award from the Leukemia and Lymphoma Society.

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Authors and Affiliations

  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., D2-373, Seattle, 98109, Washington, USA

    Barbara Varnum-Finney, Carolyn Brashem-Stein, Cynthia Nourigat, David Flowers & Irwin D. Bernstein

  2. Department of Pathology and Institute for Medicine and Engineering, 611 BRB II/III, 421 Curie Blvd.

    Lanwei Xu, Sonia Bakkour & Warren S. Pear

  3. University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Lanwei Xu, Sonia Bakkour & Warren S. Pear

  4. The Department of Pediatrics, The University of Washington, Seattle, 98105, Washington, USA

    Irwin D. Bernstein

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  1. Barbara Varnum-Finney
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  2. Lanwei Xu
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Correspondence to Barbara Varnum-Finney.

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Varnum-Finney, B., Xu, L., Brashem-Stein, C. et al. Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling. Nat Med 6, 1278–1281 (2000). https://doi.org/10.1038/81390

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