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

被引：21

作者：

Barrier, Marianne ^{[1
]}

Jeffay, Susan ^{[1
]}

Nichols, Harriette P. ^{[1
]}

Chandler, Kelly J. ^{[1
,2
]}

Hoopes, Maria R. ^{[1
]}

Slentz-Kesler, Kimberly ^{[1
]}

Hunter, E. Sidney, III ^{[1
]}

机构：

[1] US EPA, ORD, NHEERL, ISTD,SBB,MD 72, Res Triangle Pk, NC 27711 USA

[2] US EPA, ORD, NHEERL, NCCT, Res Triangle Pk, NC 27711 USA

来源：

REPRODUCTIVE TOXICOLOGY | 2011年 / 31卷 / 04期

关键词：

ACDC; mESC; Mouse embryonic stem cell; EST; Embryonic stem cell test; Developmental toxicology; Xenobiotic; MHC; VITRO EMBRYOTOXICITY TESTS; IN-VITRO; DEVELOPMENTAL TOXICITY; PREDICTION MODELS; GENE-EXPRESSION; VALIDATION; PREVALIDATION; CULTURE; PHASE;

D O I：

10.1016/j.reprotox.2011.01.007

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

An adherent cell differentiation and cytotoxicity (ACDC) assay was developed using pluripotentil 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. TagMan 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. (C) 2011 Elsevier Inc. All rights reserved.

引用

页码：383 / 391

页数：9

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