A simple short term method to study thyroid disruption using a fetal rat thyroid culture.

ABSTRACT

INTRODUCTION: Thyroid modulation activity has not been investigated for many chemical substances. Due to ethical, practical and financial reasons, in vivo evaluation of a large number of compounds is not feasible. It has been proposed that an in vitro mechanism-based strategy could be more adequate for the identification of thyroid hormone disrupting chemicals. Here we describe a simple and mostly inexpensive, short term culture assay to study thyroid disruption. METHODS: Fetal thyroids collected from gestation day 20.5 were cultured up to 24h in Hank's saline solution, at 37°C with oxygenation at 0 and 12h. Viability of the cultured explants was evaluated by the MTT assay. Positive (thyroid stimulating hormone, TSH) and negative (6-propyl-2-thiouracil, PTU) modulation of cultured thyroids was assessed with morphometrical analysis of H & E stained gland sections. Thyroxine expression was evaluated by immunohistochemistry. RESULTS: Viability was shown to increase with time of culture with higher metabolic activity being achieved at 24h as compared to shorter periods of incubation. Follicular epithelial cells exhibited a statistically significant dependence on thyrotropin concentration, although more evident in the inner than in the outer portion of the glands. As expected, TSH induced expression of thyroxin while PTU inhibited it. DISCUSSION: GD20.5 fetal thyroids may be cultured up to 24h under relatively simple laboratory conditions during which viability and function of the gland are preserved showing that it is possible to reproduce in vivo response under in vitro conditions. This culture could be a suitable short term assay to study mechanism of thyroid disruption.