US EPA, ORD, NHEERL, ISTD, SBB, MD-72, Research Triangle Park, NC 27711, USA.
Reprod Toxicol. 2011 May;31(4):383-91. doi: 10.1016/j.reprotox.2011.01.007. Epub 2011 Feb 3.
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.
采用多能 J1 小鼠胚胎干细胞 (mESC) 建立了黏附细胞分化和细胞毒性 (ACDC) 测定法。黏附 mESC 用于评估化学诱导物对细胞活力和分化的影响，方法是在 9 天的培养后使用细胞内 Western 技术进行检测。DRAQ5/Sapphire700 染色用于定量细胞数量。肌球蛋白重链蛋白用作心肌细胞分化的标志物，并对细胞数量进行校正，从而区分细胞毒性和分化作用。使用胚胎干细胞试验和 ACDC 测定法评估乙酸、5-氟尿嘧啶和溴氯乙酸。这两个系统都能区分这些化合物的相对效力。TaqMan 低密度阵列用于表征分化的时间过程以及化学暴露对多个分化基因标志物的影响。ACDC 测定法是一种可用于评估外源化学物质对 mESC 分化和细胞数量影响的技术，可通过单次测定进行。
