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Development of Hematopoietic Stem and Progenitor Cells from Mouse Embryonic Stem Cells, In Vitro, Supported by Ectopic Human HOXB4 Expression

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Embryonic Stem Cell Immunobiology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1029))

Abstract

Differentiation of pluripotent embryonic stem (ES) cells can recapitulate many aspects of hematopoiesis, in vitro, and can even generate cells capable of long-term multilineage repopulation after transplantation into recipient mice, when the homeodomain transcription factor HOXB4 is ectopically expressed. Thus, the ES-cell differentiation system is of great value for a detailed understanding of the process of blood formation. Furthermore, it is also promising for future application in hematopoietic cell and gene therapy. Since the arrival of techniques which allow the reprogramming of somatic cells back to an ES cell-like state, the generation of hematopoietic stem cells from patient-specific so-called induced pluripotent stem cells shows great promise for future therapeutic applications. In this chapter, we describe how to cultivate a certain feeder cell-independent mouse embryonic stem cell line, to manipulate these cells by retroviral gene transfer to ectopically express HOXB4, to differentiate these ES cells via embryoid body formation, and to selectively expand the arising, HOXB4-expressing hematopoietic stem and progenitor cells.

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Acknowledgments

This work is dedicated to the memory of Wolfram Ostertag (1937–2010) and Hartmut Beug (1945–2011).

As well as Bernd Schiedlmeier, Christopher Baum (Department of Experimental Hematology, Hannover Medical School), and Bernd Giebel and Peter Horn (Institute for Transfusion Medicine, University Hospital Essen) for their steady support and continuing fruitful discussions.

This work was supported by grants of the Deutsche Forschungsgemeinschaft [KL 1311/4-2; KL1311/5-1, cluster of excellence “REBIRTH” (Exc 62)] and by the NIH (grant number RO1-HL073015-06, subaward No. 1000830668).

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Authors and Affiliations

  1. Department of Medical Biology, The Walter and Eliza Hall Institute for Medical Research, Parkville, Victoria, Australia

    Sandra Pilat

  2. Molecular Immunology Division and Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia

    Sebastian Carotta

  3. Institute for Transfusion, University Hospital Essen, Essen, Germany

    Hannes Klump

Authors
  1. Sandra Pilat
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  2. Sebastian Carotta
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  3. Hannes Klump
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Editor information

Editors and Affiliations

  1. Dept. Internal Medicine, Div. Allergy & Immunology, University of Iowa, Hawkins Drive 200, Iowa City, 52242, Iowa, USA

    Nicholas Zavazava

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Pilat, S., Carotta, S., Klump, H. (2013). Development of Hematopoietic Stem and Progenitor Cells from Mouse Embryonic Stem Cells, In Vitro, Supported by Ectopic Human HOXB4 Expression. In: Zavazava, N. (eds) Embryonic Stem Cell Immunobiology. Methods in Molecular Biology, vol 1029. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-478-4_10

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  • DOI: https://doi.org/10.1007/978-1-62703-478-4_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-477-7

  • Online ISBN: 978-1-62703-478-4

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