{"title":"Preparation of Human Primary Colon Tissue-Derived Organoid Using Air Liquid Interface Culture","authors":"Tatsuya Usui, Masashi Sakurai, Koji Umata, Hideyuki Yamawaki, Takashi Ohama, Koichi Sato","doi":"10.1002/cptx.40","DOIUrl":null,"url":null,"abstract":"<p>In vitro analysis of intestinal epithelium has been hindered by a lack of suitable culture systems useful for gastrointestinal research. To overcome the problem, an air liquid interface (ALI) method using a collagen gel was established to culture three-dimensional primary cells containing both primary epithelial and mesenchymal components from mouse gastrointestinal tissues. ALI organoids accurately recapitulate organ structures, multilineage differentiation, and physiology. Since ALI organoids from human tissues have not been produced, we modified the previous protocol for mouse ALI organoid culture to establish the culture system of ALI organoids from normal and tumor colorectal tissues of human patients. The current unit presents a protocol for preparation of the ALI organoid culture from normal and tumor colorectal tissues of human patients. ALI organoid culture from human tissues might be useful for examining not only resistance to chemotherapy in a tumor microenvironment but also toxic effects on organoids. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"75 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.40","citationCount":"19","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cptx.40","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 19
Abstract
In vitro analysis of intestinal epithelium has been hindered by a lack of suitable culture systems useful for gastrointestinal research. To overcome the problem, an air liquid interface (ALI) method using a collagen gel was established to culture three-dimensional primary cells containing both primary epithelial and mesenchymal components from mouse gastrointestinal tissues. ALI organoids accurately recapitulate organ structures, multilineage differentiation, and physiology. Since ALI organoids from human tissues have not been produced, we modified the previous protocol for mouse ALI organoid culture to establish the culture system of ALI organoids from normal and tumor colorectal tissues of human patients. The current unit presents a protocol for preparation of the ALI organoid culture from normal and tumor colorectal tissues of human patients. ALI organoid culture from human tissues might be useful for examining not only resistance to chemotherapy in a tumor microenvironment but also toxic effects on organoids. © 2018 by John Wiley & Sons, Inc.
气液界面培养制备人原代结肠组织类器官
肠上皮的体外分析由于缺乏适合胃肠道研究的培养系统而受到阻碍。为了克服这一问题,建立了一种使用胶原凝胶的气液界面(ALI)方法来培养小鼠胃肠道组织中含有原代上皮和间充质成分的三维原代细胞。ALI类器官准确地概括了器官结构、多谱系分化和生理学。由于尚未从人体组织中产生ALI类器官,我们修改了先前的小鼠ALI类器官培养方案,建立了从人类患者正常和肿瘤结肠组织中提取ALI类器官的培养体系。目前,本单位提出了一种从人类患者的正常和肿瘤结肠组织中制备ALI类器官培养的方案。来自人体组织的ALI类器官培养不仅可用于检查肿瘤微环境中对化疗的耐药性,还可用于检查对类器官的毒性作用。©2018 by John Wiley &儿子,Inc。
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