HISTOCULTURE OF SKIN FOR TOXICITY ASSAY

描述：（改编自申请人的摘要）有一个重要的 局部应用和环境毒理学研究的需要 用于体外系统的试剂，以研究其对皮肤的毒性作用。 的in 体外系统具有允许研究大量各种 在单个样本上的物质。 一种体外系统， 皮肤的成分允许研究确定药剂对皮肤的影响， 各种细胞类型和细胞相互作用。 这是 特别是在三维组织培养系统中的情况。 的 申请人开发了凝胶支撑的三维组织培养系统 允许完整的生长和毒性测试的所有组成部分， 小鼠皮肤，包括角质形成细胞、真皮成纤维细胞和毛囊 细胞以及毛发，持续10天或更长时间（Li，L.，马戈利斯， L.B.和霍夫曼，R.M. Proc. Natl. Acad. 科学，In Press）. 是 这里提出，使用组织培养系统和荧光染料， 测量细胞活力或细胞死亡，以研究 以一系列农药、有机锡和致癌物为模型化合物， 对各种组织培养小鼠皮肤毒性和致癌性研究 和人类包皮以及从培养的皮肤上生长的毛发。 的 毒素的影响的测量大大增强了共聚焦 显微镜，允许三维观察标本 以前是不可能的。 至少一种化合物的体外作用 已经在体内使用无胸腺裸鼠进行了相关性研究， 体外细胞毒性效应对应于这些的刺激指数 各种化合物在体内评分。

DESCRIPTION: (Adapted from applicant's abstract) There is an important need in the study of toxicology of topically-applied and environmental agents for an in vitro system to study their toxic effects on skin. The in vitro systems have the benefit of allowing study of a large variety of substances on an individual specimen. An in vitro system containing all the components of skin allows studies to determine the effects of agents on the various component cell types and cell interactions. This is particularly the case in three-dimensional histoculture systems. The applicant developed a gel-supported, three-dimensional histoculture system that allows the intact growth and toxicity testing of all components of mouse skin, including keratinocytes, dermal fibroblasts and hair follicle cells as well as hair, for periods of 10 days or more (Li, L., Margolis, L.B. and Hoffman, R.M. Proc. Natl. Acad. Sci., In Press). It is proposed here, using the histoculture system and fluorescent dyes that measure cell viability or cell death, to investigate the effects of a series of pesticides, organotins and carcinogens as model compounds to study toxicity and carcinogenesis on all types of histocultured mouse skin and human foreskin as well as on hair growth from the cultured skin. The measurement of the effects of the toxins are greatly enhanced by confocal microscopy which allows the three-dimensional observation of the specimens not previously possible. The effects of at least one compound in vitro have already correlated in vivo using athymic nude mice such that the cytotoxic effects in vitro correspond to an irritation index for these various compounds scored in vivo.