#MILTENYI TEXMACS SKIN#
The two skin compartments develop over several gestational periods ( Ersch and Stallmach, 1999 Coolen et al., 2010). Skin development is driven by a mutual inductive mechanism between ectoderm (epidermis) and mesoderm (dermis) ( Carlson, 2012). The developing conceptus is, in principle, protected from pathogens by the uterine barrier and maternal-derived antibodies while establishing a functioning immune network. hair follicle cycling, wound repair), but they also can cause inflammation and autoimmune diseases ( Cruz et al., 2018 Ho and Kupper, 2019). Multiple T cell subsets are involved in the defense against pathogens and tumors, and play a role in tissue homeostasis (e.g. Work over the last two decades has highlighted their importance in adult human skin, which harbors twice as many T cells as are seen in the circulation ( Clark et al., 2006b). T cells, as key actors of the adaptive immune system, are commonly identified by CD3 expression, and detect antigens through heterodimeric T cell receptors (TCRs) composed of either α and β or γ and δ chains ( Morath and Schamel, 2020). Regulation of both innate and adaptive skin immunity is essential in preserving host integrity, thereby preventing inappropriate immune activation and pathology. There, a complex network of cellular and molecular pathways is employed, which allows the immune system to respond quickly and efficiently to harmful stimuli, while largely ignoring innocuous substances. The 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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