I-Corps: Human toxicity assay using synthetic embryo-like structures

这个I-Corps项目更广泛的影响/商业潜力是开发一种可控和可扩展的微流体技术，其中合成的人类胚胎样结构可以来自人类干细胞，用于定量毒性测试。所提出的技术的开发解决了对具有成本效益和高度预测性的测定平台的需求，以确定胚胎毒理学效应。本试验使用动物模型如大鼠或小鼠进行。这些动物试验成本高昂，存在伦理问题，预测能力有限。拟议中的技术提供了与人类相关的“生物体级”毒性测试。 该方法允许定量测量和扰动，并与实时成像和高通量屏幕兼容。所提出的技术可以使用疾病和患者特异性干细胞进行疾病建模，并且还可以允许使用来自不同年龄和种族/民族的干细胞进行筛选，从而模拟不同人群对药理学和化学化合物的反应。这个I-Corps项目是基于可控和可扩展的微流体技术的发展，其中合成胚胎-来源于人干细胞的类似结构可用于胚胎毒性测试。所提出的微流体装置允许将人干细胞集落精确定位在微流体装置内的规定位置。 此外，所提出的装置允许精确的时间递送可溶性因子，以驱动这些人类干细胞集落发育并组织成多细胞结构，其分子和细胞特征与早期植入后人类胚胎具有显著的相似性。所提出的技术已被证明是可控的，可重复的，可扩展的，并与活成像和免疫细胞化学兼容，这可能允许容易地采用到现有的高通量毒性筛选管道。该技术旨在提供基于比色/荧光试剂的药理化合物对胚胎发育影响的高含量表型和形态分析。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this I-Corps project is the development of a controllable and scalable microfluidic technology in which synthetic human embryo-like structures may be derived from human stem cells for quantitative toxicity testing. The development of the proposed technology addresses the need for a cost-effective and highly predictive assay platform to determine embryo toxicology effects. The current assay is conducted using animal models such as rats or mice. These animal tests are costly and have ethical issues and limited predictive powers. The proposed technology offers human-relevant, “organism-level” toxicity testing. The method allows quantitative measurements and perturbations and is compatible with live imaging and high-throughput screens. The proposed technology may use disease- and patient-specific stem cells for disease modeling, and also may allow screening using stem cells derived from different age and racial/ethnic groups, thus modeling responses to pharmacological and chemical compounds of diverse populations.This I-Corps project is based on the development of a controllable and scalable microfluidic technology in which synthetic embryo-like structures derived from human stem cells may be utilized for embryo toxicity testing. The proposed microfluidic device allows precise positioning of human stem cell colonies in prescribed locations within the microfluidic device. In addition, the proposed device allows precise temporal delivery of soluble factors to drive these human stem cell colonies to develop and organize into multicellular structures whose molecular and cellular features bear significant similarities to early post-implantation human embryos. The proposed technology has been shown to be controllable, reproducible, scalable, and compatible with live imaging and immunocytochemistry, which may allow easy adoption into the existing high-throughput toxicity screening pipelines. The technology is designed to provide high-content phenotypic and morphological profiling of the effects of pharmacological compounds on embryo development based on colorimetric/fluorescent reagents.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.