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- W4378388051 abstract "Background: Modelling atopic dermatitis (AD) in vitro is paramount to understand the disease pathophysiology and identify novel treatments. Previous studies have shown that the Th2 cytokines IL-4 and IL-13 induce AD-like features in keratinocytes in vitro . However, it has not been systematically researched whether the addition of Th2 cells, their supernatants or a 3D structure are superior to model AD compared to simple 2D cell culture with cytokines. Methods: For the first time, we investigated what in vitro option most closely resembles the disease in vivo based on single-cell RNA sequencing data (scRNA-seq) obtained from skin biopsies in a clinical study and published datasets of healthy and AD donors. In vitro models were generated with primary fibroblasts and keratinocytes, subjected to cytokine treatment or Th2 cell cocultures in 2D/3D. Gene expression changes were assessed using qPCR and Multiplex Immunoassays. Results: Of all cytokines tested, incubation of keratinocytes and fibroblasts with IL-4 and IL-13 induced the closest in vivo -like AD phenotype which was observed in the scRNA-seq data. Addition of Th2 cells to fibroblasts failed to model AD due to the downregulation of ECM-associated genes such as POSTN. While keratinocytes cultured in 3D showed better stratification than in 2D, changes induced with AD triggers did not better resemble AD keratinocyte subtypes observed in vivo . Conclusions : Taken together, our comprehensive study shows that the simple model using IL-4 or IL-13 in 2D most accurately models AD in fibroblasts and keratinocytes in vitro , which may aid the discovery of novel treatment options." @default.
- W4378388051 created "2023-05-27" @default.
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- W4378388051 date "2023-05-26" @default.
- W4378388051 modified "2023-10-18" @default.
- W4378388051 title "Single-cell RNA sequencing reveals 2D cytokine assay can model atopic dermatitis more accurately than immune-competent 3D setup" @default.
- W4378388051 doi "https://doi.org/10.22541/au.168507561.19506037/v1" @default.
- W4378388051 hasPublicationYear "2023" @default.
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