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Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity (ACDC) assay-Book Chapter*
Citation:
BARRIER, M., K. J. CHANDLER, S. C. JEFFAY, M. Hoopes, T. Knudson, AND E. S. HUNTER. Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity (ACDC) assay-Book Chapter*. Chapter 11, J. Hansen & C. Harris (ed.), Developmental Toxicity: Series: Methods in Molecular Biology. Humana Press Incorporated, Totowa, NJ, 889:181-195, (2012).
Impact/Purpose:
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 following a 9-day culture period.
Description:
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 following a 9-day culture period. DRAQ5/Sapphire700 stains are used to quantify cell number and an In-Cell Western technique is used to measure myosin heavy chain (MHC) protein immunostaining that is a marker of cardiomyocyte differentiation. MHC protein content 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.