An immunoresponsive three-dimensional urine-tolerant human urothelial model to study urinary tract infection

• DOI: 10.3389/fcimb.2023.1128132
• Publication Date: 2023-03-27T06:25:46Z
• Authors: Nazila V. Jafari; Jennifer L. Rohn
• Keywords: Urothelial Cell; Barrier function

Introduction Murine models of urinary tract infection (UTI) have improved our understanding host-pathogen interactions. However, given differences between rodent and human bladders which may modulate host bacterial response, including certain biomarkers, urothelial thickness the concentration urine, development new human-based is important to complement mouse studies provide a more complete picture UTI in patients. Methods We originally developed three-dimensional (3D) model was urine tolerant demonstrated several but it only achieved heterogenous, multi-layered zones did not demonstrate comprehensive differentiation status needed achieve barrier function. optimised this by altering variety conditions validated with microscopy, flow cytometry, transepithelial electrical resistance FITC-dextran permeability assays confirm tissue architecture, integrity response infection. Results an 3D urine-tolerant (3D-UHU), after 18-20 days growth, stratified uniformly 7-8 layers comprised three expected, distinct cell types. The apical surface differentiated into large, CD227+ umbrella-like cells expressing uroplakin-1A, II, III, cytokeratin 20, all are terminal markers, glycosaminoglycan layer. Below layer, intermediate were present, single underlying layer CD271+ basal cells. also expressed E-cadherin, ZO-1, claudin-1 -3, possessed good Infection both Gram-negative Gram-positive classes elicited elevated levels pro-inflammatory cytokines chemokines characteristic humans caused decrease Discussion Taken together, 3D-UHU holds promise for studying interactions immune response.