Reproductive Toxicology 2011-05-01

Mouse embryonic stem cell adherent cell differentiation and cytotoxicity (ACDC) assay.

Marianne Barrier, Susan Jeffay, Harriette P Nichols, Kelly J Chandler, Maria R Hoopes, Kimberly Slentz-Kesler, E Sidney Hunter

文献索引：Reprod. Toxicol. 31(4) , 383-91, (2011)

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摘要

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.Copyright © 2011 Elsevier Inc. All rights reserved.