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Differentiation and Lineage Commitment of Murine Embryonic Stem Cells into Insulin Producing Cells

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Embryonic Stem Cell Immunobiology

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

Abstract

Pluripotent embryonic stem (ES) cells and induced pluripotent stem (iPS) cells recently developed in our laboratory can be used to generate the much needed insulin producing cells (IPCs) for the treatment of type 1 diabetes. However, currently available differentiation protocols generate IPCs at a very low frequency. More importantly, it is difficult to purify the IPCs from the mixed cell population due to the lack of well characterized pancreatic beta cell-specific cell surface markers. Subsequently, multiple studies have been published with limited success. A major cause for these poor results is an inadequate Pdx1 expression in the embryoid body (EB) or definitive endoderm (DE)-derived precursors. Here we investigated whether ectopic expression of pancreatic and duodenal homeobox 1 (Pdx1), an essential pancreatic transcription factor, in mouse ES cells leads to enhanced differentiation into IPCs. Here we describe a new approach for the generation of glucose responsive IPCs using ES cells ectopically expressing pancreatic and duodenal homeobox 1 (Pdx1) and paired box gene 4 (Pax4).

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Acknowledgements

This study was made possible by Grant Number 1I01BX001125-02, NIH/NHLBI #R01 HLO73015 and a VA Merit Review Award to N.Z. and by an NIH Pilot Grant to S.R. from the University of Iowa, Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, Grant Number NIH/NIDDK #P30 DK054759. S.R. is a recipient of the 2009 Young Investigator Award by the American Society of Transplantation.

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Authors and Affiliations

  1. Department of Internal Medicine, Division of Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA

    Sudhanshu P. Raikwar

  2. Veterans Affairs Medical Center, Iowa City, IA, USA

    Sudhanshu P. Raikwar & Nicholas Zavazava

  3. Department of Internal Medicine and VAMC Iowa City, Division of Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, USA

    Nicholas Zavazava

  4. Immunology Graduate Program, University of Iowa, Iowa City, IA, USA

    Nicholas Zavazava

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  1. Sudhanshu P. Raikwar
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  2. Nicholas Zavazava
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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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Raikwar, S.P., Zavazava, N. (2013). Differentiation and Lineage Commitment of Murine Embryonic Stem Cells into Insulin Producing Cells. 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_7

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  • DOI: https://doi.org/10.1007/978-1-62703-478-4_7

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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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