# Structural and Viability Assessment of Bovine Corneas Preserved at an Accessible Low-Temperature (−20 °C) for Eye Irritation Models

## TL;DR

This paper explores a low-cost method to preserve bovine corneas at -20°C for use in eye irritation testing, making the process more accessible for labs with limited resources.

## Contribution

The study introduces a practical low-temperature preservation protocol using 10% DMSO at -20°C for bovine corneas.

## Key findings

- Storage at -20°C with 10% DMSO preserved structural morphology and transparency effectively.
- The protocol is cost-effective and suitable for labs with limited infrastructure.
- Preservation outcomes were evaluated using corneal transparency, histology, and endothelial viability.

## Abstract

In vitro ocular irritation testing requires
the
use of freshly excised bovine corneas, which limits broader adoption
in laboratories with restricted access to slaughterhouse material.
This study investigated a practical and accessible low-temperature
preservation strategy focused on maintaining structural and cellular
integrity. Bovine corneas were stored at −20 or −80
°C using two commonly available cryoprotective agents: dimethyl
sulfoxide (DMSO) and ethylene glycol (EG). Preservation outcomes were
assessed through corneal transparency, histological architecture,
and endothelial viability. Among the tested conditions, storage at
−20 °C with 10% DMSO provided the most consistent preservation
of structural morphology and transparency. This protocol provides
a cost-effective foundation for extending the usability of excised
corneas and supports future efforts to expand the accessibility of in vitro eye irritation models in laboratories with limited
infrastructure.