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Effects Of Haloacetic Acids and their major metabolites in a Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity (ACDC) Assay
Citation:
JEFFAY, S. C., H. P. NICHOLS, M. BARRIER, AND E. S. HUNTER. Effects Of Haloacetic Acids and their major metabolites in a Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity (ACDC) Assay. Presented at Society of Toxicology Meeting, Salt Lake City, UT, March 07 - 11, 2010.
Impact/Purpose:
The haloacetic acids (HAAs) are a class of chemicals produced by disinfection of drinking water
Description:
The haloacetic acids (HAAs) are a class of chemicals produced by disinfection of drinking water. Many of the HAAs are developmental toxicants when administered to rodents producing a variety of developmental effects. We have previously shown that the HAAs can produce direct effects on rodent embryonic development in whole embryo culture. These studies evaluated the effects of individual HAAs and their major metabolites on mouse embryonic stem cell development using the adherent cell differentiation/cytotoxicity method (ACDC). The HAAs evaluated were iodoacetic acid (MIA), chloroacetic (MCA), fluoroacetic acid (MFA), bromoacetic (MBA), dichloroacetic (DCA), dibromoacetic (DBA), bromochloroacetic (BCA), trichloroacetic (TCA), tribromoacetic (TBA), bromodichloroacetic (BDCA) and dibromochloroacetic (DBCA) acids. The metabolites evaluated were glyoxylic acid (GOA), oxylic acid (OXA), glycolic acid (GCA). Acetic acid (AA) was also evaluated. A single cell suspension ofpluripotent 11 mouse embryonic stem cells (mESC) were plated in 96-well plates and maintained under differentiating conditions for 9 days. The concentration dependent effects of each compound on cell number (DRAQ5/Sapphire700 staining) and differentiation (quantitative in-cell Western analysis ofmyosin heavy chain protein) were evaluated. The concentration that produced a 50% reduction was calculated for each endpoint. For cytotoxicity the relative potencies of the compounds were (listed from most to least potent) MIA>MBA>BCA=MCA>DBA>GOA>MFA>TBA>TCA>DBCA=BDCA>AA=DCA=GCA.Whenilie concentrations that produced an effect on differentiation were compared to those that were cytotoxic: BCA, DBCA and DCA were more potent at altering differentiation than inducing cytotoxicity; MIA, MBA, GOA, MFA were more potent cytotoxicants than at altering differentiation; MCA, DBA, TBA, TCA, BDCA produced effects on both endpoints at similar concentrations. AA and GCA produced no effects at concentrations less than or equal to 10mM. Based on previous results, although the absolute concentrations required to alter embryonic development in whole embryo culture are different than in stem cells, the relative potency ofthe HAAs are similar in the two systems. Thus, many HAAs alter differentiation or perturb cell proliferation or induce excess cell death in mouse embryonic stem cells. [This abstract does not reflect EPA policy