Complex in vitro Systems, Safety Assessment, Genentech, 1 DNA Way, South San Francisco, CA, 94080, USA.
Sci Rep. 2023 Jun 27;13(1):10412. doi: 10.1038/s41598-023-35657-9.
Primary tissue-derived epithelial organoids are a physiologically relevant in vitro intestinal model that have been implemented for both basic research and drug development applications. The existing method of culturing intestinal organoids in surface-attached native extracellular matrix (ECM) hydrogel domes is not readily amenable to large-scale culture and contributes to culture heterogeneity. We have developed a method of culturing intestinal organoids within suspended basement membrane extract (BME) hydrogels of various geometries, which streamlines the protocol, increases the scalability, enables kinetic sampling, and improves culture uniformity without specialized equipment or additional expertise. We demonstrate the compatibility of this method with multiple culture formats, and provide examples of suspended BME hydrogel organoids in downstream applications: implementation in a medium-throughput drug screen and generation of Transwell monolayers for barrier evaluation. The suspended BME hydrogel culture method will allow intestinal organoids, and potentially other organoid types, to be used more widely and at higher throughputs than previously possible.
原代组织衍生的上皮类器官是一种具有生理相关性的体外肠道模型，已被应用于基础研究和药物开发。目前，在附着于表面的天然细胞外基质（ECM）水凝胶穹顶中培养肠道类器官的方法不容易进行大规模培养，并且导致培养的异质性。我们开发了一种在各种形状的悬浮基底膜提取物（BME）水凝胶中培养肠道类器官的方法，该方法简化了方案，增加了可扩展性，实现了动力学采样，并提高了培养的均一性，而无需专用设备或额外的专业知识。我们证明了该方法与多种培养形式的兼容性，并提供了悬浮 BME 水凝胶类器官在下游应用中的实例：在高通量药物筛选中的应用和用于屏障评估的 Transwell 单层的生成。悬浮 BME 水凝胶培养方法将使肠道类器官，以及其他潜在的类器官类型，能够比以前更广泛和更高通量地使用。
