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Hokkaido University Collection of Scholarly and Academic Papers >
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Efficient and simple genetic engineering of enteroids using mouse isolated crypts for investigating intestinal functions
| Title: | Efficient and simple genetic engineering of enteroids using mouse isolated crypts for investigating intestinal functions |
| Authors: | Ohira, Shuya Browse this author | | Yokoi, Yuki Browse this author | | Ayabe, Tokiyoshi Browse this author | | Nakamura, Kiminori Browse this author →KAKEN DB |
| Keywords: | Gene transfer | | Transgenic enteroid | | Isolated crypt | | Small intestinal epithelial cell | | Stem cell niche |
| Issue Date: | 31-Dec-2022 |
| Publisher: | Elsevier |
| Journal Title: | Biochemical and biophysical research communications |
| Volume: | 637 |
| Start Page: | 153 |
| End Page: | 160 |
| Publisher DOI: | 10.1016/j.bbrc.2022.11.008 |
| Abstract: | Intestinal epithelial cells separate subepithelial tissues from luminal environment formed with food, incoming pathogens, and resident intestinal microbiota, etc., and elicit various intestinal function. Enteroid, a three-dimensional culture system of small intestinal epithelial cells, has been widely used for analyzing the intestinal function, further a transgenic enteroid was developed to investigate the mo-lecular mechanisms. However, conventional transgenic enteroid production method, which transfer gene into single stem cells, has limitations including low efficiency and time-consuming. Here we show that by gene transfer into small intestinal isolated crypts maintaining stem cell niche, a transgenic enteroid was obtained quickly and efficiently. Isolated crypts were transfected by lentiviral vector without separating into single cells, and transgenic enteroid composed of all lineages of intestinal epithelial cells was generated at day 7 with yield of 56%, maintaining the intestinal function in drug transport and innate immunity. Our efficient and simple transgenic enteroid generation method enables high-throughput investigation of intestinal epithelial cells and contributes to understanding intestinal function.(c) 2022 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
| Type: | article |
| URI: | http://hdl.handle.net/2115/88895 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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