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Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity Assay

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Developmental Toxicology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 889))

Abstract

There are thousands of environmental chemicals for which there is limited toxicological information, motivating the development and application of in vitro systems to profile the biological effects of xenobiotic exposure and predict their potential developmental hazard. An adherent cell differentiation and cytotoxicity (ACDC) assay was developed using pluripotent mouse embryonic stem cells (mESCs) to evaluate chemical-induced effects on both stem cell viability and differentiation. This assay uses an In-Cell Western technique after a 9-day culture. DRAQ5/Sapphire700 cell/DNA stains are used to quantify cell number and myosin heavy chain (MHC) protein is used as a marker of cardiomyocyte differentiation. MHC is corrected for cell number, thereby separating cytotoxicity and effects on differentiation. The ACDC assay can be used to evaluate the effects of xenobiotics on mESC differentiation and cell number in the same sample.

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Author information

Authors and Affiliations

  1. National Health and Environmental Effects Research Laboratory, Research Triangle Park, NC, USA

    Marianne Barrier

  2. National Health and Environmental Effects Research Laboratory and National Center for Computational Toxicology Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA

    Kelly Chandler

  3. National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA

    Susan Jeffay & Maria Hoopes

  4. National Center for Computational Toxicology, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA

    Tom Knudsen & Sid Hunter

Authors
  1. Marianne Barrier
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  2. Kelly Chandler
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  3. Susan Jeffay
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  4. Maria Hoopes
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  5. Tom Knudsen
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  6. Sid Hunter
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Corresponding author

Correspondence to Sid Hunter .

Editor information

Editors and Affiliations

  1. Toxicology Program, Dept. Environmental Health Sciences, University of Michigan, Washington Heights 1420, Ann Arbor, 48109-2029, Michigan, USA

    Craig Harris

  2. , Department of Pediatrics, Emory University, Uppergate Drive NE., 2015Suite 350, Atlanta, 30322, Georgia, USA

    Jason M. Hansen

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© 2012 Springer Science+Business Media, LLC

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Barrier, M., Chandler, K., Jeffay, S., Hoopes, M., Knudsen, T., Hunter, S. (2012). Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity Assay. In: Harris, C., Hansen, J. (eds) Developmental Toxicology. Methods in Molecular Biology, vol 889. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-867-2_11

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  • DOI: https://doi.org/10.1007/978-1-61779-867-2_11

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-866-5

  • Online ISBN: 978-1-61779-867-2

  • eBook Packages: Springer Protocols

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