A Flow Cytometry Workflow for Identifying Myeloid Subsets in Porcine Wound Tissue

Myeloid cells, particularly macrophages, and granulocytes, are central regulators of the inflammatory and reparative phases of wound healing. Pigs represent a highly translational large-animal model for studying cutaneous injury and repair. However, methods for isolating and analysing porcine skin wound-infiltrating myeloid cells are limited. Here, we present a protocol using enzymatic digestion for the isolation of single cells with high viability from porcine skin following excision injury. Cells isolated with this method are suitable for flow cytometry and other downstream applications. Using this approach, we assessed myeloid populations across different stages of the wound healing process. This protocol can be helpful for future studies aiming at defining cellular mechanisms of inflammation and repair in porcine wound healing.