The cytokine, interleukin-33 (IL-33), has emerged as a promising therapeutic target in chronic respiratory diseases and viral exacerbations. Despite great progress being made with simple cell assays, aspects of IL-33 biology require further investigation in complex, translational systems. Consequently, the aim of this work was to develop a human precision-cut lung slice (huPCLS) platform suitable to explore IL-33 biology and test novel therapies. huPCLS were generated and cultured for up to 7 days. LDH levels and H&E staining were used to evaluate tissue viability and architecture with inflammatory cytokine release following LPS stimulation measured to assess huPCLS responsiveness. IL-33 stimulated huPCLS were analysed via bulk RNA-sequencing (RNA-seq) whilst immunohistochemistry was used to identify airway epithelial subsets. huPCLS were also exposed to human rhinovirus A (HRV-A) with cytokine release utilised as a readout of infection. LDH levels peaked 1 day post slicing and decreased with time in culture. Tissue architecture was maintained at day 7 versus day 0 whilst 24-hour LPS stimulation induced significant IL-6, IL-1? and TNF? release relative to unstimulated huPCLS. In pilot studies, IL-33 responsive genes were upregulated when IL-33-stimulated huPCLS were analysed by bulk RNA-seq. Furthermore, airway epithelial subsets known to be modulated by IL-33 could be identified in huPCLS. Finally, infection with HRV-A increased IFN? and IL-5 release by huPCLS relative to control slices. In summary, a method to generate viable and responsive huPCLS has been developed with promising initial results demonstrating the potential of the platform to investigate IL-33 biology and screen novel therapies.