015 Optimal methods for human skin T-cell analysis

Background:Tissue-resident memory T (TRM) cells reside in peripheral tissues, including the skin. In humans, TRM exist both epidermis and dermis, but their composition, phenotype, function are not fully elucidated. To analyze skin needs to be incubated with Dispase II separate two layers. The next step varies among researchers; subsequent enzymatic digestion of dermis is popular, whereas spontaneous migration method can also done. Objective: This study aimed determine strengths limitations method. Methods: Healthy, non-inflamed human was epidermal dermal sheets, which were subject either or floated on a medium obtain emigrants. Results: An enzyme combination collagenases, collagenase alone, cleaved CD4, CD8, CD69 cells. lost ∼20% floating sheets. However, there no significant bias regard CD103 expression between emigrants remaining Through method, it determined that 104 105 CD3+ per 1 cm2 respectively. Conclusions: might useful for T-cell analysis.