Human Corneal Model for Ocular Irritation Assay

用于眼刺激测定的人类角膜模型

• 批准号: 8411604
• 负责人: Yulia Kaluzhny
• 金额: $ 18.16万
• 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/8411604
• 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.

眼球刺激性试验替代兔眼刺激试验仍是一种 难以捉摸的目标。许多人认为角膜全厚度(CFT)模型是完全 建立外源化学物质与人类角膜相互作用的模型。在第一阶段，人类 从自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，冷冻保存。确定了培养条件，以产生 高度分化的2层(角膜上皮和含有角质细胞的基质)和3- 层(上皮、间质和内皮)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 建立了检测和预测模型(PM)，使CFT组织能够区分 刺激性和非刺激性之间的100%准确率。其他功能研究表明， CFT组织似乎适合用于评估恢复时间(TTR)和损伤深度(DOI)。 在化学损伤之后。最后，对CFT组织生产的分析表明， CFT的经济性是有利的。 第二阶段将在第一阶段结果的基础上进一步发展CFT组织模型。 初步目标将侧重于准备用于商业生产的CFT组织模型-细胞 将扩大库存，以确保稳定的电池供应和质量控制参数 发展起来的。使用CFT测试的材料数据库将得到扩展，PM将用于 刺激性/非刺激性分类将最终确定，实验室之间的可转移性 化验将进行演示。最后，CFT组织将用于TTR和DOI 测量和测试和预测模型将取代德雷兹测试 发展起来的。由于这些增强的能力，CFT将为毒物学家和 科学家拥有一种工具，即使不是完全消除，也将极大地减少对 动物试验，以确定眼睛刺激性。