Introduction: Skin fibrosis is a prevalent characteristic observed in autoimmune skin diseases such as psoriasis and systemic sclerosis. Despite progress in using skin biopsies to advance our understanding of these diseases, effectively profiling immune cells and pathological fibroblasts to precisely characterize the dysregulations driven by these pathologic cells poses a persistent challenge.
Methods: We have devised a comprehensive 15-color flow cytometry panel designed to immunophenotype rare tissue-resident fibroblast populations and common tissue-resident immune cells in healthy human and cynomolgus monkey dermal sections. Human dermal sections obtained from GenoSkin were shipped overnight at room temperature and cynomolgus skin samples were shipped overnight cryopreserved with CryoStor CS10 freezing medium. Both tissues were mechanically dissected and further treated with Collagenase IV enzymatic digestion or Miltenyi’s MACS Whole Skin Dissociation Kit for tissue disruption.
Results: Digestion of human punch biopsies processed with Collagenase IV produced qualitatively superior results in comparison to Miltenyi’s MACS Whole Skin Dissociation Kit. Substantial levels of antibody saturation on target receptors were retained after skin tissue disruption followed by staining with fluorescently labeled antibodies for both Collagenase IV and Miltenyi’s MACS Whole Skin Dissociation methods, however, Collagenase IV resulted in higher yield for our cells of interest.
Conclusion: We observed similar immune profiling in human and cynomolgus dermal sections and sustained levels of antibody-mediated target saturation when using both tissue disruption methodologies evaluated in this study.
