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Ex vivo expansion of human hematopoietic stem cells by direct delivery of the HOXB4 homeoprotein

Nature Medicine volume 9, pages 1423–1427 (2003)Cite this article

Abstract

Expansion of human hematopoietic stem cells (HSCs) is a major challenge in cellular therapy, and currently relies on the use of recombinant cytokines or on gene transfer of transcription factors. Of these, the HOXB4 homeoprotein protein is of particular interests as it promotes the expansion of mouse HSCs without inducing the development of leukemia. To eliminate any deleterious effects that might be associated with stable HOXB4 gene transfer into human cells, we took advantage of the ability of HOX proteins to passively translocate through cell membranes. Here we show that when cultured on stromal cells genetically engineered to secrete HOXB4, human long-term culture-initiating cells (LTC-ICs) and nonobese diabetic–severe combined immunodeficiency (NOD-SCID) mouse repopulating cells (SRCs) were expanded by more than 20- and 2.5-fold, respectively, over their input numbers. This expansion was associated with enhanced stem cell repopulating capacity in vivo and maintenance of pluripotentiality. This method provides a basis for developing cell therapy strategies using expanded HSCs that are not genetically modified.

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Figure 1: Generation and properties of HOXB4 expressing MS-5 cells.
Figure 2: HOXB4 translocates into human HSCs and binds its target DNA sequence.
Figure 3: Relative amplification of total cells, CFCs and LTC-ICs.
Figure 4: Ex vivo expansion of SRCs in coculture with MS-5/SP-HOXB4 cells.

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Acknowledgements

We thank G. Sauvageau and K. Humphries for the human HOXB4 cDNA; A. Gould for the I12 monoclonal antibody to HOXB4; A. Prochiantz for his sound advice; M.-C. Gendron for cell sorting; J.-M. Freyssinier, M. Talbot and M. Titeux for technical help; I. Khazaal and P. Ardouin for their help with breeding and care of the NOD-SCID mice; and I. Dusanter-Fourt, C. Francastel, F. Gesbert, S. Gisselbrecht and N. Taylor for helpful discussions and comments on the manuscript. This work was supported by grants from the Institut de la Santé et de la Recherche Médicale (INSERM) and the Association Française contre les Myopathies (no. 4CS04F). S.A. is supported by a poste d'accueil INSERM.

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  1. Département d'Hématologie, Institut Cochin, INSERM U567, CNRS UMR 8104, Université Paris 5, Maternité Port-Royal, 123, Bd de Port-Royal, Paris, 75014, France

    Sophie Amsellem, Françoise Pflumio, Dominique Bardinet, Brigitte Izac, Paul-Henri Romeo, Anne Dubart-Kupperschmitt & Serge Fichelson

  2. Unité d'Oncologie Virale, Institut Pasteur, 25–28 rue du Dr Roux, Paris, 75015, France

    Pierre Charneau

  3. AP-HP, Hôpital Cochin, 27 rue du Fg St Jacques, Paris, 75014, France

    Serge Fichelson

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  1. Sophie Amsellem
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Correspondence to Serge Fichelson.

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Amsellem, S., Pflumio, F., Bardinet, D. et al. Ex vivo expansion of human hematopoietic stem cells by direct delivery of the HOXB4 homeoprotein. Nat Med 9, 1423–1427 (2003). https://doi.org/10.1038/nm953

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