A 2D Intestinal Crypt Platform for Compound Screens

项目摘要 肠道上皮是人体内分裂最快的组织，细胞的健康状况可以 容易受到环境毒素和药物的影响。因此，这个器官系统是一种质数 药用化合物、益生元和微生物产品的生物测试目标。An In 能够准确预测这些影响的体外肠道模型将通过以下方式减轻测试负担 减少对动物模型的依赖，最大限度地减少重叠评估的数量 帮助消除不必要的测试。为了满足这一需求，Altis BiosSystems Inc. 生物技术公司，将与华盛顿大学的科学家合作开发 一种新的二维(2D)隐窝平台，模拟肠道上皮。人类原始性 肠上皮干细胞将在平台上形成图案，以模拟肠上皮 具有干细胞利基，具有迁移性增殖和分化细胞。此平面2D 平台将以坚固和可重复的方式实现高含量/高通量分析。 在此阶段I SBIR中，此协作将生成包含 分化细胞单层，优化平台，重点放在量化策略上 对性能进行基准测试，并进行小规模筛选，以演示推进的可行性 添加到第二阶段项目。目标是开发和基准一种定量的体外方法，该方法将 使研究人员能够有效地评估化合物对肠道干细胞的影响 增殖分化。

Project Summary Intestinal epithelium is the fastest dividing tissue in the body, and the health of the cells can be easily impacted by environmental toxins and drugs. This organ system is therefore a prime target for biological testing of medicinal compounds, prebiotics and microbial products. An in vitro gut model that can precisely predict these impacts will reduce the burden of testing by diminishing reliance on animal models, minimizing the number of overlapping assessments and helping to eliminate unnecessary testing. To meet this need, Altis Biosystems Inc., an early stage biotechnology company, will collaborate with scientists at University of Washington to develop a novel 2-dimensional (2D) crypt platform that emulates the gut epithelium. Human primary intestinal epithelial stem cells will be patterned on the platform to mimic a gut epithelium possessing a stem-cell niche, and migratory proliferating and differentiating cells. This planar 2D platform will enable high-content/high-throughput assays in a rugged and reproducible manner. In this Phase I SBIR, this collaboration will generate 2D crypts with intervening regions of a differentiated cell monolayer, optimize the platform with a focus on quantitative strategies to benchmark performance, and perform small scale screens to demonstrate feasibility to advance to a Phase 2 project. The goal is to develop and benchmark a quantitative in vitro method that will enable investigators to efficiently evaluate the impact of compounds on intestinal stem cell proliferation and differentiation.