Co-culture with intestinal epithelial organoids allows efficient expansion and motility analysis of intraepithelial lymphocytes

• DOI: 10.1007/s00535-016-1170-8
• Publication Date: 2016-01-22T08:14:33Z
• Authors: Kengo Nozaki; Wakana Mochizuki; Yuka Matsumoto; Taichi Matsumoto; Masayoshi Fukuda; Tomohiro Mizutani; Mamoru Watanabe; Tetsuya Nakamura
• Keywords: Organoid; Intraepithelial lymphocyte; Surgical oncology; Colorectal Surgery

Abstract Background Intraepithelial lymphocytes (IELs) in the intestine play important roles regulation of local immune responses. Although their functions have been studied a variety animal experiments, vitro studies on spatiotemporal behaviors IELs and interaction with intestinal epithelial cells (IECs) hampered due to lack suitable culture system. In this study, we aimed at developing novel co-culture system IECs investigate dynamic between these two populations vitro. Methods We optimized experimental conditions under which murine can be efficiently maintained cultured as three-dimensional organoids. then tested effect IL-2, IL-7, IL-15 maintenance By time-lapse imaging, also examined IELs. Results expanded organoids presence IL-15. were within outside showing ~four-fold increase both αβT γδT for period 2 weeks. Four-dimensional fluorescent imaging revealed an active, multi-directional movement along basolateral surface IECs, inward or outward migration relative organoid structures. Cell tracking analysis showed that shared indistinguishable features regard dynamics. Conclusions This method could serve unique tool motility dynamics temporal spatial