title: Mouse embryonic stem cell adherent cell differentiation and cytotoxicity (ACDC) assay.
abstract: An adherent cell differentiation and cytotoxicity (ACDC) assay was developed using pluripotent J1 mouse embryonic stem cells (mESCs). Adherent mESCs were used to evaluate chemical-induced effects on both stem cell viability and differentiation using an in-cell western technique after a 9-day culture. DRAQ5/Sapphire700 stains were used to quantify cell number. Myosin heavy chain protein was used as a marker of cardiomyocyte differentiation and was corrected for cell number, thereby separating cytotoxicity and effects on differentiation. Acetic acid, 5-fluorouracil and bromochloroacetic acid were evaluated using the embryonic stem cell test and ACDC assay. Both systems distinguish the relative potencies of these compounds. TaqMan low-density arrays were used to characterize the time course of differentiation and effects of chemical exposure on multiple differentiation gene markers. The ACDC assay is a technique that can be used to evaluate the effects of xenobiotics on mESC differentiation and cell number using a single assay.
authors.affiliation: US EPA, ORD, NHEERL, ISTD, SBB, MD-72, Research Triangle Park, NC 27711, USA. Barrier.Marianne@epa.gov
authors.forename: Marianne
authors.initials: M
authors.lastname: Barrier
authors.forename: Susan
authors.initials: S
authors.lastname: Jeffay
authors.forename: Harriette P
authors.initials: HP
authors.lastname: Nichols
authors.forename: Kelly J
authors.initials: KJ
authors.lastname: Chandler
authors.forename: Maria R
authors.initials: MR
authors.lastname: Hoopes
authors.forename: Kimberly
authors.initials: K
authors.lastname: Slentz-Kesler
authors.forename: E Sidney
authors.initials: ES
authors.lastname: Hunter
mesh_terms: D000085:Acetates / Q000633:toxicity; D019342:Acetic Acid / Q000633:toxicity; D000826:Animal Testing Alternatives; D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000473:pathology; D005472:Fluorouracil / Q000633:toxicity; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051379:Mice; D009200:Myocardial Contraction / Q000187:drug effects* / Q000235:genetics; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000187:drug effects* / Q000473:pathology; D016133:Polymerase Chain Reaction; D018570:Risk Assessment; D013997:Time Factors; D018675:Toxicity Tests
chemical_list: D000085:Acetates; C099813:bromochloroacetic acid; D019342:Acetic Acid; D005472:Fluorouracil