(WHS-P77) Establishment of a protocol for multicolor flow cytometry of porcine skin cells
Friday, May 17, 2024
7:30 AM – 5:00 PM East Coast USA Time
Due to their similarity to human skin, pigs are increasingly used to study wound healing. Both pig and human skin have firm attachment, sparse hair coat, thick epidermis and dermis, no panniculus carnosus, and heal by re-epithelialization with minimal contraction. Flow cytometry is a robust method to analyze the inflammatory milieu at the wound site. Although efforts have been made to optimize porcine skin digestion for flow cytometric analysis, it is currently not a common practice and there are no widely used protocols. Our lab optimized a method to isolate single cells from porcine skin following excision injury using an enzymatic digestion that yield consistently robust single cell isolation with high viability. We also optimized reliable antibody panels, for the identification of granulocytes (CD45+2B2+), monocytes subsets (CD45+2B2-SLADR-CD14+CD16+/-), as well as M1 inflammatory (CD45+2B2-SLADR+CD80+) and M2 reparative (CD45+2B2-SLADR+CD163+) macrophages. Development of a reliable method to isolate single cells from porcine skin opens the door for studies to assess the cellular changes during porcine wound healing.