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The Validated Embryonic Stem Cell Test with Murine Embryonic Stem Cells

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Teratogenicity Testing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1797))

Abstract

Birth defects are the leading cause of infant mortality in the USA, yet the causes of most of these conditions are unknown. While a combination of genetic and environmental factors are suspected in most cases, little information exists about the health risks that prenatal exposure to many common chemicals poses for the fetus. Thus, development and refinement of procedures that can accurately predict embryotoxicity of compounds is important for curtailing the number of infants born with birth defects. The embryonic stem cell test (EST) is a procedure that utilizes comparison of cytotoxicity in embryonic and adult cells and inhibition of differentiation to predict embryotoxicity of compounds tested. Because of its use of existing cell lines, the EST dramatically reduces the need for animal test subjects in toxicity testing. In addition, because of its use of inhibition of differentiation as an endpoint, the EST is extremely versatile in the range of complications it can test for. In this chapter, procedures for use of the validated embryonic stem cell test with the traditional cardiomyocyte differentiation endpoint are explained. The protocol includes discussion of routine stem cell culture, the cardiomyocyte differentiation procedure, and methods for utilization of molecular endpoints for assessing embryotoxicity of compounds.

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Acknowledgments

This chapter was written with support from the National Institutes of Health, National Institute of Dental and Craniofacial Research (R01DE025330-01A1), and the Tobacco Related Disease Research Program (#25IP-0018) to N.z.N. and a National Science Foundation Graduate Research Fellowship to L.M.W.

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Authors and Affiliations

  1. Department of Molecular, Cell and Systems Biology, College of Natural and Agricultural Sciences, University of California Riverside, Riverside, CA, USA

    Lauren M. Walker & Nicole I. zur Nieden

  2. Division of Natural Sciences, Department of Biology, El Camino College, Torrance, CA, USA

    Darcie L. V. McClelland-Descalzo

  3. Stem Cell Center, College of Natural and Agricultural Sciences, University of California Riverside, Riverside, CA, USA

    Nicole I. zur Nieden

Authors
  1. Lauren M. Walker
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  2. Darcie L. V. McClelland-Descalzo
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  3. Nicole I. zur Nieden
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Corresponding author

Correspondence to Nicole I. zur Nieden .

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Editors and Affiliations

  1. Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal

    Luís Félix

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Walker, L.M., McClelland-Descalzo, D.L.V., zur Nieden, N.I. (2018). The Validated Embryonic Stem Cell Test with Murine Embryonic Stem Cells. In: Félix, L. (eds) Teratogenicity Testing. Methods in Molecular Biology, vol 1797. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7883-0_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7883-0_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7882-3

  • Online ISBN: 978-1-4939-7883-0

  • eBook Packages: Springer Protocols

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