Human-Derived Precision-Cut Lung Slices (hPCLS): Agarose Filling, Coring and Slicing Protocol v1

Human Precision-Cut Lung Slices (hPCLS) are uniform tissue slices generated from human lungs. These slices contain all resident lung cells and their interactions, including alveolar macrophages and dendritic cells, present in the tissue at the time of slicing, representing the natural complexity of the lung’s 3-dimensional architecture and organ function physiology. hPCLS has become one of the most relevantex vivoorganotypic models for studying the pathophysiology of human lungs. Thanks to specialized tissue slicers, a single lung can provide hundreds of slices for both control and experimental conditions. Additionally, hPCLS can be kept in culture for up to 4 weeks, making possible to mimic the onset and progression of lung diseases. hPCLS has proven valuable in multiple applications such as RNA and protein analysis, biochemistry, metabolism, toxicology, and chronic and acute lung diseases. They have also been used for highly specialized downstream applications such as transcriptomics, proteomics and high-resolution live microscopy. The following protocol is a guide on how to generate PCLS from human lungs.