Cultivation and characterisation of primary and subcultured equine keratinocytes.

The research discusses the development and analysis of equine keratinocyte cultures sustained with a high proliferative capacity up to their second passage. These are valuable when studying the biology of equine keratinocytes and the origin of skin disorders. The study also contributes to solving wound healing issues in equine dermatology.

Methodology

• The team created and detailed equine keratinocyte cultures. These are cells that produce keratin, a protein essential for the protection and repair of skin tissues.
• The cultures were maintained in a high proliferation capacity up to the second passage, meaning that they were capable of rapid growth and replication for extended periods.
• Improved attachment and growth were achieved by placing primary cells on equine feeder layers, suggesting that these cells perform best in their natural environment.
• Subcultured keratinocytes, cells propagated under lab conditions, showed optimal growth when seeded on collagen type I, outperforming those seeded on equine feeder layers in terms of their proliferation rate.

Findings

• Through immunohistochemistry (a technique used to visually see and localize antigens in cells of tissue), the origins and state of differentiation of the equine keratinocytes were determined.
• These cultured cells expressed keratin and desmoplakin I/II (key molecules in cell adhesion), but lacked keratin 10, showing that they were of a specific genetic makeup suitable for specific investigations into skin health.
• Observation through electron microscopy revealed ordinary features of cultured keratinocytes, which suggests that these cells retained their normal characteristics even in a lab environment.
• The purity of keratinocyte cultures was determined by vimentin staining. Vimentin is a type of intermediate filament (IF) protein that is often used as a marker of mesenchymal (connective tissue) cells.

Significance

• This is the first report on the establishment of equine keratinocytes derived from lip epithelium and forms the foundation to study equine keratinocyte biology and the pathogenesis of epidermal diseases.
• Given that wound healing is a major issue in equine dermatology, this research may be crucial for the development of new and improved therapies.