Protocol for Immune Cell Isolation, Organoid Generation, and Co-culture Establishment from Cryopreserved Whole Human Intestine

从冷冻保存的整个人类肠道中分离免疫细胞、生成类器官和建立共培养物的方案

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作者:Enrique Gamero-Estevez, Inga Viktoria Hensel, Michelle Steinhauer, Olivia Müllertz, Elizaveta Savochkina, Ibrahim Murathan Sektioglu, Bilgenaz Stoll, Shaghayegh Derakhshani, Sarah Devriese, Kyungbo Kim, Martin Resnik-Docampo

Abstract

The human intestine plays a pivotal role in nutrient absorption and immune system regulation. Along the longitudinal axis, cell-type composition changes to meet the varying functional requirements. Therefore, our protocol focuses on the processing of the whole human intestine to facilitate the analysis of region-specific characteristics such as tissue architecture and changes in cell populations. We describe how to generate a biobank that can be used to isolate specific immune cell subtypes, generate organoid lines, and establish autologous immune cell-organoid co-cultures. Key features • Dissection and tissue analysis of whole human intestines. • Cryopreservation for biobank generation. • Optimized protocols for the isolation of epithelial and immune cells. • Autologous co-culture of organoids and lamina propria-derived immune cells.

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