Innovative three-dimensional testicular Co-culture (Mini-Testis) model for reproductive toxicity testing: a pathway based High throughput (HT) and High Content Analysis (HCA)

用于生殖毒性测试的创新三维睾丸共培养（迷你睾丸）模型：基于途径的高通量（HT）和高内涵分析（HCA）

基本信息

批准号：
9198371
负责人：
Lei Yin
金额：
$ 20.21万
依托单位：
REPROTOX BIOTECH, LLC
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2018-01-31
项目状态：
已结题

项目摘要

Summary The need to identify chemicals for their potential to cause reproductive and developmental (R/D) toxicity (R/D) is a significant public health challenge for toxicologists. In vivo testing for reproductive and developmental toxicity assessment uses a large number of animals, yet these studies are criticized for their inefficiency. The National Research Council (NRC)’s report “Toxicity Testing in the 21st Century” envisioned that in vivo animal testing will eventually be replaced by a combination of in silico and in vitro approaches. Therefore, the need to identify in vitro systems for reproductive and developmental toxicity testing has grown. ReproTOX has established an animal-free testicular cell co-culture system (Mini-Testis) from testicular cell lines. Both morphology and cellular biomarkers confirmed that this animal-free 3D-co-culture model support in vitro spermatogenesis. Our 32-compound preliminary study revealed a stronger correlation of IC50 from the Mini-Testis model with in vivo reproductive toxicity as compared to any single testicular cell culture model. These initial results suggest that our Mini-Testis model might be a valuable screening tool for reproductive toxicants and prioritizing chemicals for further testing. The specific aims of this proposal, therefore, are to (1) establish a toxicity-based high throughput Mini-Testis model for R/D toxicity evaluation and (2) develop an integrated pathway-based high content (HCA) and high throughput (HT) screening assay in the Mini-Testis model for R/D toxicity evaluation. The project outcome will be a cost-efficient, pathway-based in vitro Mini- Testis model for potential R/D toxicity screening of chemicals.

概括需要识别化学物质，以引起复制和发育（R/D）毒性的潜力（R/D）对于毒理学家来说是一个重大的公共卫生挑战。体内测试生殖和发育毒性评估使用了大量动物，但是这些研究对它们而言至关重要效率低下。国家研究委员会（NRC）的报告“ 21世纪的毒性测试”设想体内动物测试最终将被硅和体外方法的组合所取代。因此，鉴定体外系统以进行生殖和发育毒性测试的需求已经增长。 reprotox已建立了一个来自检测细胞的动物的检测共培养系统（MINI-TESTIS）线。形态和细胞生物标志物都证实了这种无动物的3D-CO培养模型支持体外精子发生。我们的32个复合初步研究表明，IC50的相关性更强与任何单个验证细胞培养模型相比，具有体内生殖毒性的迷你泰式模型。这些最初的结果表明，我们的迷你泰式模型可能是生殖的有价值的筛选工具有毒物质和优先级化学物质进行进一步测试。因此，该提议的具体目的是（1）建立基于毒性的高吞吐量微型泰式模型，用于R/D毒性评估，（2）基于途径的高素质高素质（HCA）和高吞吐量（HT）筛选分析 R/D毒性评估的模型。该项目结果将是一个基于经济高效的，基于途径的体外微型睾丸模型，用于对化学物质的潜在毒性筛选。