Human Corneal Model for Ocular Irritation Assay

用于眼刺激测定的人类角膜模型

• 批准号: 8209216
• 负责人: Yulia Kaluzhny
• 金额: $ 39.08万
• 依托单位: MATTEK CORPORATION
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209216
• 关键词: Address; Adherent Culture; Animal Testing; Animal Welfare; Animals; Biological Assay; Cells; Chemical Injury; Chemicals; Classification; Cornea; Corneal Endothelium; Data; Databases; Development; Economics; Endothelial Cells; Epithelial; Epithelial Cells; Epithelium; Evaluation; Exhibits; Exposure to; Eye; Eye Injuries; Generations; Goals; Harvest; Human; In Vitro; Industry; Injury; Irritants; Laboratories; Materials Testing; Measurement; Methods; Modeling; Oryctolagus cuniculus; Pathology; Performance; Pharmacologic Substance; Phase; Phenotype; Physiology; Production; Proliferating; Quality Control; Recovery; Reliance; Reproducibility; Safety; Scientist; Stromal Cells; Testing; Thick; Time; Tissue Model; Tissues; Variant; Wound Healing; base; corneal epithelium; experience; in vitro Assay; in vitro testing; in vivo Model; irritation; research study; response; time use; tool

Replacement of the Draize rabbit eye test for ocular irritancy testing remains an elusive goal. Many believe a corneal full thickness (CFT) model is necessary to completely model the interaction of exogenous chemicals with human cornea. During Phase 1, human corneal epithelial, stromal, and endothelial cells were isolated from native tissue, expanded in monolayer culture, and cryopreserved. Culture conditions were identified to produce highly differentiated 2-layer (corneal epithelium and keratocyte-containing stroma) and 3- layer (epithelium, stroma, and endothelium) CFT tissues. The CFT tissues exhibited improvements over epithelial tissues in terms of barrier and differentiated phenotype. An assay and prediction model (PM) was developed which allowed the CFT tissue to distinguish between irritants and non-irritants with 100% accuracy. Other functional studies showed that the CFT tissue appears suitable to evaluate time to recovery (TTR) and depth of injury (DOI) following chemical injury. Finally, an analysis of CFT tissue production showed that the economics of CFT are favorable. Phase II will expand upon the Phase 1 result to further develop the CFT tissue model. Initial goals will focus on preparing the CFT tissue model for commercial production - cell stocks will be expanded to insure a stable cell supply and quality control parameters will be developed. The database of materials tested using the CFT will be expanded, the PM for irritant/non-irritant classification will be finalized, and inter-laboratory transferability of the assay will be demonstrated. Finally, the CFT tissues will be used for TTR and DOI measurements and an assay and prediction model for replacement of the Draize test will be developed. Due to these enhanced capabilities, the CFT will provide toxicologists and scientists with a tool that will dramatically decrease, if not entirely eliminate, the need for animal testing to determine ocular irritancy.

用Draize兔眼试验代替眼刺激试验仍然是一种可行的方法。 难以捉摸的目标许多人认为，角膜全层（CFT）模型是必要的， 模拟外源性化学物质与人类角膜的相互作用。在第1阶段，人类 从天然组织中分离角膜上皮细胞、基质细胞和内皮细胞，扩增 单层培养，并冷冻保存。确定了产生 高度分化的2层（角膜上皮和含角质细胞的基质）和3层 层（上皮、基质和内皮）CFT组织。CFT组织显示 在屏障和分化表型方面优于上皮组织。一个 分析和预测模型（PM）的发展，使CFT组织区分 刺激物和非刺激物之间100%的准确性。其他功能研究表明， CFT组织似乎适用于评估恢复时间（TTR）和损伤深度（DOI） 化学伤害后。最后，对CFT组织生产的分析表明， CFT的经济性是有利的。 II期将在1期结果的基础上进行扩展，以进一步开发CFT组织模型。 最初的目标将集中在制备用于商业生产的CFT组织模型-细胞 将扩大库存，以确保稳定的细胞供应， 开发使用CFT测试的材料数据库将得到扩展， 将最终确定刺激性/非刺激性分类，并确定 检测将被证明。最后，CFT组织将用于TTR和DOI 测量和测定和预测模型的替代Draize测试将是 开发由于这些增强的能力，CFT将提供毒理学家和 科学家们有一个工具，将大大减少，如果不是完全消除，需要 动物试验以确定眼睛刺激性。