Department of Biomedical Engineering, Tufts University, Medford, Massachusetts.
Department of Physics and Astronomy, Tufts University, Medford, Massachusetts.
Curr Protoc. 2024 Sep;4(9):e70013. doi: 10.1002/cpz1.70013.
The study of human intestinal physiology and host-microbe interactions is crucial for understanding gastrointestinal health and disease. Traditional two-dimensional cell culture models lack the complexity of the native intestinal environment, limiting their utility in studying intestinal biology. Here, we present a detailed protocol for the set up and utilization of a three-dimensional (3D) in vitro bioreactor system for human intestinal studies and bacterial co-culture. This article outlines the design and assembly of the bioreactor system, scaffold fabrication, bacterial culture techniques, analysis methods, and troubleshooting tips. By providing step-by-step instructions, the goal is to enable other laboratories to utilize physiologically relevant tissue models of the human intestine, incorporating key features, such as nutrient flow, multiple human cell types, 3D architecture, and microbial communities. The incorporation of commensal bacteria into the bioreactor system allows for the investigation of complex host-microbe interactions, providing insight into gastrointestinal health and pathology. This article serves as a comprehensive resource for scientists seeking to advance their understanding of intestinal biology toward the development of novel therapeutic strategies for gastrointestinal disorders. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Scaffold design Basic Protocol 2: Intestinal cell culture: Caco2 cells Basic Protocol 3: Intestinal cell culture: organoids Basic Protocol 4: Bioreactor design and set up Basic Protocol 5: Bacteria in 3D bioreactor set up Basic Protocol 6: Bacteria and drug dosing.
研究人类肠道生理学和宿主-微生物相互作用对于理解胃肠道健康和疾病至关重要。传统的二维细胞培养模型缺乏天然肠道环境的复杂性，限制了它们在研究肠道生物学中的应用。本文介绍了一种用于人类肠道研究和细菌共培养的三维（3D）体外生物反应器系统的详细设置和使用方案。本文概述了生物反应器系统的设计和组装、支架制造、细菌培养技术、分析方法以及故障排除技巧。通过提供逐步的说明，目标是使其他实验室能够利用具有生理相关性的人类肠道组织模型，纳入关键特征，如营养流动、多种人类细胞类型、3D 架构和微生物群落。将共生细菌纳入生物反应器系统可以研究复杂的宿主-微生物相互作用，深入了解胃肠道健康和病理学。本文为寻求深入了解肠道生物学以开发胃肠道疾病新治疗策略的科学家提供了全面的资源。© 2024 威立出版社。基础方案 1：支架设计基础方案 2：Caco2 细胞的肠道细胞培养基础方案 3：类器官的肠道细胞培养基础方案 4：生物反应器设计和设置基础方案 5：3D 生物反应器中细菌的设置基础方案 6：细菌和药物剂量。
