Development of a microphysiologic system to assay the interaction of the human colonic epithelium on Clostridium difficile

项目概要 在当前的申请中，提出了一个协作的、多学科的研究项目 开发并验证一个创新平台来研究原始人类之间的相互作用 肠道细胞和病原微生物艰难梭菌（C. difficile）。一个新颖且独特的 工程技术将创建一个肠芯片（器官芯片）平台，能够 维持与腔内非致病性和共培养的存活的人类结肠上皮 致病性专性厌氧菌，以研究发生的基本细胞过程 在人类和微生物细胞相互作用的过程中。这项工作将设计、建模和 设计一个支持生理学的三维肠道细胞培养平台 粘液层并模仿粪便物质通过的纯粹力量。的设计 该系统将使观察和扰动人类微生物细胞细胞成为可能 相互作用并修改各自的环境以阐明潜在的细胞和 分子事件是这些相互作用的关键。该提案的一个强大属性是，这些 实验将利用新的干细胞培养方法来产生正常的、原代的 人类结肠上皮而不是使用肿瘤细胞系作为哺乳动物细胞的来源。 所提出的微生理系统生成的实验数据将在未来 提供有关艰难梭菌预防和治疗方案的关键信息以及 为研究其他感染性生物体提供了一种新技术 胃肠道。

Project Summary In the current application, a collaborative, multidisciplinary research project is proposed to develop and validate an innovative platform to study the interactions between primary human intestinal cells and the pathogenic microbe, Clostridium difficile (C. difficile). A new and uniquely engineered technology will create an intestine-on-chip (organ-on-chip) platform capable of maintaining a viable human colonic epithelium co-cultured with luminal nonpathogenic and pathogenic obligate anaerobes in order to study fundamental cellular processes that occur during the interaction between the human and microbial cells. The work will design, model and engineer a three-dimensional intestinal-cell culture platform that supports a physiologic mucous layer as well as mimics the sheer forces of the passage of fecal material. The design of this system will make it possible to observe and perturb the human-microbe cell-cell interactions and modify respective environments to elucidate the underlying cellular and molecular events key to these interactions. A powerful attribute of the proposal is that these experiments will leverage new stem-cell culture methods that produce a normal, primary human colonic epithelium rather than use tumor cell lines as the source of the mammalian cells. The experimental data generated by the proposed microphysiological system will in the future provide key information concerning prevention and treatment option(s) for C. difficile as well as provide a new technique for the study of other infectious organisms involving the gastrointestinal tract.