Project Objectives

• Standardise 2D adrenal and pancreatic cell line models for endocrine‑disrupting chemical (EDC) screening.
• Develop 3D spheroid and organoid models using high‑throughput 5D plate technology.
• Validate in‑vitro assays with chemicals of known endocrine‑disrupting activity.
• Generate primary adrenal and pancreatic 3D cultures using bovine tissue obtained from slaughterhouse waste.
• Establish protocols enabling adoption of these models across research laboratories.

3Rs Impact

• Provides in‑vitro models capable of replacing a substantial portion of rodent‑based endocrine‑disrupting chemical (EDC) testing.
• Uses slaughterhouse by‑products to generate physiologically relevant primary organoids.
• High‑throughput platforms reduce the number of necessary confirmatory in‑vivo experiments.
• Enhances scientific translatability by using human‑relevant endocrine pathways.
• Supports regulatory commitments to reduce and eventually replace animal‑based chemical safety assessments.

Background

Endocrine‑disrupting chemicals (EDCs) are natural or synthetic substances found in products such as plastics, cosmetics, detergents, pesticides, and food packaging. These chemicals can interfere with hormonal signalling and have been associated with metabolic disease, obesity, infertility, early puberty, cardiovascular disease, and various cancers. As EDC exposure is widespread and often unavoidable, identifying harmful compounds early and reliably is essential for public health.

Current EDC screening heavily depends on rodent studies, despite meaningful differences between rodent and human endocrinology, particularly in steroidogenesis and glucose regulation. To address this limitation, the project develops human‑relevant 2D and 3D in‑vitro models of the adrenal gland and pancreas. These include well‑established cell lines such as NCI‑H295R and the more recently developed MUC‑1, which uniquely reflects androgenic pathways not captured by traditional models. The project also generates high‑throughput spheroids and organoids, including those derived from bovine tissue collected from slaughterhouse waste, enabling more complex and physiologically relevant screening systems.

By creating robust, standardised platforms suitable for high‑throughput EDC testing, this project aims to provide reliable alternatives to animal experiments and support international regulatory efforts to reduce in‑vivo chemical testing.