A 2D Intestinal Crypt Platform for Compound Screens

项目摘要 肠上皮细胞是体内分裂最快的组织，细胞的健康可以 很容易受到环境毒素和药物的影响。因此，这个器官系统是一个质数。 用于药物化合物、益生元和微生物产品的生物学测试的目标。的in 体外肠道模型，可以精确地预测这些影响将减少测试的负担， 减少对动物模型的依赖，减少重叠评估的数量， 以避免不必要的测试。为了满足这一需求，Altis Biosystems Inc.，早期阶段 生物技术公司，将与华盛顿大学的科学家合作， 一种新型的二维（2D）隐窝平台，其模拟肠上皮。人原代 肠上皮干细胞将在平台上形成图案以模拟肠上皮 具有干细胞小生境，以及迁移性增殖和分化细胞。这种平面2D 该平台将以坚固和可重现的方式实现高含量/高通量测定。 在第一阶段SBIR中，这种合作将产生2D隐窝，其中插入了一个 分化的细胞单层，优化平台，重点是定量策略， 基准性能，并进行小规模筛选，以证明推进的可行性 第二阶段的项目。我们的目标是开发和基准的定量体外方法， 使研究人员能够有效地评估化合物对肠道干细胞的影响， 增殖和分化。

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.