Developmental Toxicology pp 181-195

Part of the Methods in Molecular Biology book series (MIMB, volume 889) | Cite as

Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity Assay

  • Marianne Barrier
  • Kelly Chandler
  • Susan Jeffay
  • Maria Hoopes
  • Tom Knudsen
  • Sid Hunter
Protocol

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.

Key words

ACDC mESC Mouse embryonic stem cell Developmental toxicology Xenobiotic MHC Myosin heavy chain Cardiomyocyte 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Marianne Barrier
    • 1
  • Kelly Chandler
    • 2
  • Susan Jeffay
    • 3
  • Maria Hoopes
    • 3
  • Tom Knudsen
    • 4
  • Sid Hunter
    • 4
  1. 1.National Health and Environmental Effects Research LaboratoryResearch Triangle ParkUSA
  2. 2.National Health and Environmental Effects Research Laboratory and National Center for Computational Toxicology Office of Research and DevelopmentU.S. Environmental Protection AgencyResearch Triangle ParkUSA
  3. 3.National Health and Environmental Effects Research LaboratoryU.S. Environmental Protection AgencyResearch Triangle ParkUSA
  4. 4.National Center for Computational Toxicology, Office of Research and DevelopmentU.S. Environmental Protection AgencyResearch Triangle ParkUSA

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