2/20/2023 0 Comments Texmacs t cell isolationThe immune system constantly surveys the microenvironment and discriminates between harmless and potentially harmful components – a multifaceted task at barrier surfaces, such as the skin, that are constantly exposed to exogenous stimuli. Collectively, our experimental approaches and data advance the understanding of fetal skin immunity and potential use in future therapeutic interventions. Importantly, culture of total fetal skin biopsies facilitated T cell expansion without a substantial impact on their phenotype, a major prerequisite for subsequent functional assays. Their molecular state was characterized using single-cell transcriptomics and TCR repertoire profiling. Using multiparametric flow cytometry and in situ immunofluorescence, we found a large population with a naive phenotype and small populations with a memory and regulatory phenotype. Here, we describe a new workflow for isolating and expanding significant amounts of T cells from fetal human skin. T cells are first observed in the skin at the early stages of gestation however, our understanding of their contribution to early immunity has been limited by their low abundance and lack of comprehensive methodologies for their assessment. The adult human skin contains a vast number of T cells that are essential for skin homeostasis and pathogen defense.
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