Gut-associated lymphoid tissues (GALTs) comprise key intestinal immune inductive sites, including the Peyer’s patches of the small intestine and different types of isolated lymphoid follicle (ILF) found along the length of the gut. Our understanding of human GALT is limited due to a lack of protocols for their isolation. Here we describe a technique that, uniquely among intestinal cell isolation protocols, allows identification and isolation of all human GALT, as well as GALT-free intestinal lamina propria (LP). The technique involves the mechanical separation of intestinal mucosa from the submucosa, allowing the identification and isolation of submucosal ILF (SM-ILF), LP-embedded mucosal ILF (M-ILF) and LP free of contaminating lymphoid tissue. Individual SM-ILF, M-ILF and Peyer’s patch follicles can be subsequently digested for downstream cellular and molecular characterization. The technique, which takes 4–10 h, will be useful for researchers interested in intestinal immune development and function in health and disease.
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We thank all patients and collaborating staff at Herlev hospital and, in particular, the Gastroenterology Team (Department of Pathology) for help in providing tissue samples. We thank A. Mowat (Glasgow University) for valuable feedback during preparation of the manuscript and J. Vandamme for video editing assistance. This work was supported by grants from the Lundbeck Foundation (R155-2014-4184), Denmark, the Gut Cell Atlas, an initiative funded by the Leona M. and Harry B. Helmsley Charitable Trust, US, the Swedish Research Council (2017-02072) and the Swedish Cancerfonden (18 0598) to W.W.A.
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