Teratogenic effects of two known teratogens (Nicotine and Cadmium) and prevention of such effects by addition of antioxidants in chick embryos: An evaluation of two culture systems (Micromass and in ovo culture)

Abstract: This study aimed to evaluate effectiveness of the chick cardiomyocyte micromass (MM) system for prediction of teratogenecity of common environmental factors and prevention of such effects by addition of multivitamins. White Leghorn 5-day-old embryo hearts were dissected and disassociate to produce a cardiomyocyte cell suspension in Dulbecco's Modified Eagle's Medium. The cultures were incubated at 370C in 5% CO2 in air. Microscopic observations regarding cell beating and contractility were made at 24, 48 and 144 hours post culture. Cellular viability/metabolic activity was assessed by using the resazurin reduction assay and cell total protein content was assessed by the kenacid blue assay. It was observed that cadmium chloride induced toxicity was not reversed by addition of folic acid and vitamin C, however the nicotine induced teratogenic effects were reversed by addition of folic acid and vitamin C. The results demonstrate the potential of the chick cardiomyocyte MM culture assay to identify teratogens/embryotoxins that alter morphology and function, which may result in either teratogenic outcome or causing cytotoxicity and also protection brought about by addition of multivitamins. This could form part of a screen for developmental toxicity related to cardiac function. The effects of nicotine were compared to in ovo culture also showed protective effects of multivitamins in nicotine induced teratogenecity.