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An Ex vivo Culture System to Study Thyroid Development
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A simple short term method to study thyroid disruption using a fetal rat thyroid culture.

Pablo Olivares-Trigo1, Camilo Fuentes Peña1, María Fernanda Cavieres1

  • 1Escuela de Química y Farmacia, Facultad de Farmacia, Universidad de Valparaíso, Valparaíso, CHILE, Av Gran Bretaña 1093, Playa Ancha, Valparaíso, Chile.

Journal of Pharmacological and Toxicological Methods
|May 7, 2017
PubMed
Summary

This study presents a simple, inexpensive in vitro assay using fetal thyroid glands to assess thyroid disruption. The method preserves gland viability and function for up to 24 hours, proving effective for identifying thyroid hormone disrupting chemicals.

Keywords:
Alternative methodsMTTThyroid disruptionThyroid gland culture

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Area of Science:

  • Endocrinology
  • Toxicology
  • Developmental Biology

Background:

  • Investigating thyroid modulation activity of numerous chemicals is limited by ethical and financial constraints of in vivo studies.
  • An in vitro mechanism-based strategy is proposed as a more suitable approach for identifying thyroid hormone disrupting chemicals.
  • This study introduces a novel, cost-effective short-term culture assay for thyroid disruption evaluation.

Purpose of the Study:

  • To develop and validate a simple, inexpensive in vitro short-term culture assay for studying thyroid disruption.
  • To assess the viability and function of cultured fetal thyroid glands over a 24-hour period.
  • To determine the assay's capability in reproducing in vivo responses to thyroid modulators.

Main Methods:

  • Fetal thyroid glands (gestation day 20.5) were cultured for up to 24 hours.
  • Explant viability was assessed using the MTT assay.
  • Thyroid stimulating hormone (TSH) and 6-propyl-2-thiouracil (PTU) were used to modulate thyroid function, with assessments via morphometrical analysis and immunohistochemistry for thyroxine expression.

Main Results:

  • Thyroid gland viability and metabolic activity increased with culture time, peaking at 24 hours.
  • Follicular epithelial cells showed a concentration-dependent response to thyrotropin.
  • TSH treatment significantly induced thyroxine expression, while PTU inhibited it, consistent with in vivo effects.

Conclusions:

  • Cultured GD20.5 fetal thyroids maintain viability and function for up to 24 hours under simple laboratory conditions.
  • This in vitro assay successfully reproduces in vivo responses to thyroid modulators.
  • The developed short-term culture assay is a suitable method for studying the mechanisms of thyroid disruption.