Establishment and Characterization of a Canine Keratinocyte Organoid Culture System

The study established a 3D in vitro culture system of canine keratinocyte organoids to investigate skin diseases in dogs, particularly those affecting the interfollicular epidermis (IFE) and hair follicles (HF). Skin biopsies from euthanized dogs without skin abnormalities were used to grow organoids, which developed into a basal IFE-like cell type. Gene and protein analysis showed high levels of keratins 5 and 14, but lacked key HF stem cell markers. The organoids demonstrated potential to differentiate further upon withdrawal of growth factors. The system provided a foundation for exploring epidermal function and addressing canine cutaneous disorders, though additional growth factors or co-culture with dermal papilla would be necessary for HF signatures or hair growth.