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Membrane-bound Delta-4 Notch ligand reduces the proliferative activity of primitive human hematopoietic CD34+CD38low cells while maintaining their LTC-IC potential

Leukemia volume 18, pages 788–797 (2004)Cite this article

Abstract

To examine the role of the Notch ligand Delta-4 on hematopoietic stem cells, human CD34+CD38low cord blood cells were cocultured on S17 cells transduced with transmembrane Delta-4 (mbD4/S17) or an empty vector (C/S17). By the end of a 3-week culture, mbD4/S17 induced a 25-fold reduction in nucleated cell production, as compared to C/S17, by maintaining a higher proportion of cells in G0/G1 phase. A specific retention of a high proportion of CD34+ cells throughout the culture was observed with mbD4/S17, contrary to C/S17. Although mbD4/S17 promoted expansion of cells with the phenotype of committed lymphoid precursors (CD34+CD7+CD45RA+), these cells still retained their myeloid differentiation potential. mbD4/S17 maintained a higher LTC-IC frequency in output CD34+ cells, compared to C/S17, as in the subsets of cells having completed the same number of divisions on mbD4/S17. A Delta4-Fc protein (extracellular part of human Delta4 fused to Fc human IgG1 portion), immobilized on plastic, also reduced cell production and retained the LTC-IC potential. Transplantation of cells grown on mbD4/S17 into NOD/SCID mice showed no significant enhancement of the long-term repopulating ability. Thus, Delta4 appears to inhibit hematopoietic stem cell proliferation, in association with the maintenance of short-term lymphoid and myeloid repopulation capacity.

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Acknowledgements

We thank midwifes and nurses who helped us to collect cord blood samples. We are indebted to AMGEN (Thousand Oaks, CA, USA) for providing us with rhu-SCF, KIRIN (Tokyo, Japan) for rhu-PEG-MDGF, IMMUNEX for Flt3-ligand, Novartis for rhu-IL-3, CILAG for rhuEPO, and Dr K Mori (Niigata University, Niigata, Japan) for the MS-5 cell line, and Dr Dorshkind for providing us with S17 cells. We thank Dr J Morgenstern and Dr G Laramee for providing the retroviral constructs, V Schiavon and Y Lécluse for performing the cell sorting, M Jandrot-Perrus for allowing us to purify Delta4-Fc in her laboratory, P Ardouin for mice facilities. This work was supported by INSERM, and a grant from CRC (Contrat de Recherche Clinique, CRC no. 2000.10, Institut Gustave Roussy), and a grant from ARC (Association de recherche contre le Cancer, no. 4300).

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Authors and Affiliations

  1. U362 INSERM - PR1, Institut Gustave Roussy, 39 rue C. Desmoulins, Villejuif, France

    E Lauret, C Catelain, M Titeux, S Poirault, J S Dando, A Groseil, W Vainchenker, F Sainteny & A Bennaceur-Griscelli

  2. Department of Clinical Biology, Institut Gustave Roussy, Villejuif, France

    A Bennaceur-Griscelli

  3. Inflammation Department, Millennium Pharmaceuticals Inc., 45 Sidney Street, Cambridge, 02139, MA, USA

    M Dorsch & J-L Villeval

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  1. E Lauret
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Lauret, E., Catelain, C., Titeux, M. et al. Membrane-bound Delta-4 Notch ligand reduces the proliferative activity of primitive human hematopoietic CD34+CD38low cells while maintaining their LTC-IC potential. Leukemia 18, 788–797 (2004). https://doi.org/10.1038/sj.leu.2403288

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