Generating an Ex Vivo Ovine Wounded Skin Model Infected with a Bacterial Pathogen

Take a pre-sterilized skin biopsy section obtained from a lamb's forehead in an antibiotic-free medium.

The section comprises an outer epidermis and an underlying dermis layer.

Remove the medium and use a buffer to wash the section.

Make a circular cut in the section and remove the top layer. This breaks the barrier, generating a wound bed in the skin.

Transfer the wounded sections onto a permeable membrane insert in a multiwell plate containing an antibiotic-free medium.

Introduce a suspension of pathogenic bacteria onto the wound bed and incubate.

During incubation, the wound bed provides a tissue surface for bacterial attachment and the nutrients to support bacterial growth.

This allows the multiplication and spreading of the bacteria across the wound bed, establishing an infection.