Effects of developmental toxic chemicals on mouse embryonic stem cell-derived cardiomyocytes

Program: Abstracts - Orals, Featured Poster Presentations, and Posters
Session: SUN 366-382-Physiological Impacts of Endocrine Disrupting Chemicals
Sunday, June 16, 2013: 1:45 PM-3:45 PM
Expo Halls ABC (Moscone Center)

Poster Board SUN-381
Yeoul Choi and Eui-Bae Jeung*
College of Veterinary Med, Cheongju Chungbuk, South Korea
In vitro screening assays for developmental toxicity are highly needed to increase efficiency and to reduce animal use. Embryonic stem (ES) cells are unique as they have the potential to be generated in large numbers and the ability to differentiate into the three germ layers via embryonic body (EB) formation. In this study, we investigated the effects of several developmental toxic chemicals on the differentiation of mouse embryonic stem cells (mESC) into cardiomyocyte. We used six well-known chemicals, ascorbic acid, cytosine arabinoside, dexamethasone, hydroxyurea, indomethacin, 5-fluorouracil and negative control penicillin G in mESC to test developmental toxicity. mESC was exposed to low- , medium- , and high-doses of the chemicals. mESCs were cultivated in hanging drops to form EBs. We performed quantitative real-time PCR analysis for cardiomyocyte-specific markers in cardiomyocyte during culture period (5+0, 5+3, 5+7, 5+11 days). In the beating cells observed after plating, beating frequency was monitored and evaluated daily depending on the treatment. We demonstrated that there was reduction of EB size after high doses of cytosine arabinoside, dexamethasone and hydroxyurea. Each stage of differentiation in mESCs demonstrated expression of cardiac specific gene by real-time PCR. The expression of cardiac specific genes in the toxicant treated groups was significantly different from the control. We also observed variation of beating rates from the mESC-derived cardiomyocyte. These results demonstrate that mESCs-derived cardiomyocyte can be used as an in vitro model to test developmental toxicity.

Nothing to Disclose: YC, EBJ

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