Kosim Kinga, Schilt Iris, Lanz Henriëtte L, Vulto Paul, Kurek Dorota
Mimetas BV, Leiden, The Netherlands.
Methods Mol Biol. 2022;2373:87-105. doi: 10.1007/978-1-0716-1693-2_6.
The study of epithelial barrier properties in the human body is of paramount interest to a range of disciplines, including disease modeling, drug transport studies, toxicology, developmental biology, and regenerative biology. Current day in vitro studies largely rely on growing epithelial cells in a static environment on membrane cell culture inserts. With the advancement of microfluidic and organ-on-a-chip techniques it became possible to culture 3D intestinal tubules directly against an extracellular matrix (ECM) under flow and without the need for artificial membranes. Here we describe detailed protocols for culturing epithelial tubules in a high-throughput format, assessing their permeability and marker expression. The platform harbors 40 independent microfluidic chips in a microtiter plate format. The resulting 40 epithelial tubules are analyzed in parallel using a high-content microscopy. Protocols described here allow for adoption and routine application of microfluidic techniques by nonspecialized end-users.
人体上皮屏障特性的研究对一系列学科至关重要，包括疾病建模、药物转运研究、毒理学、发育生物学和再生生物学。目前的体外研究很大程度上依赖于在膜细胞培养插入物的静态环境中培养上皮细胞。随着微流控和芯片器官技术的进步，现在可以在流动条件下直接在细胞外基质（ECM）上培养3D肠小管，而无需人工膜。在这里，我们描述了以高通量形式培养上皮小管、评估其通透性和标志物表达的详细方案。该平台以微孔板形式包含40个独立的微流控芯片。使用高内涵显微镜对所得的40个上皮小管进行平行分析。这里描述的方案允许非专业终端用户采用和常规应用微流控技术。
