HISTOCULTURE OF SKIN FOR TOXICITY ASSAY

描述：(改编自申请人的摘要)有一项重要的 外用和环境毒理学研究中的需要 用于体外系统研究其对皮肤的毒性作用的试剂。入内 体外系统的好处是可以研究大量不同的 个体样本上的物质。一种包含所有细胞的体外系统 皮肤成分的研究可以确定药物对皮肤的影响 各种组成细胞类型和细胞相互作用。这是 尤其是在三维组织文化系统中。这个 申请人开发了一种凝胶支持的三维组织培养系统 这使得所有成分的完整生长和毒性测试 小鼠皮肤，包括角质形成细胞、真皮成纤维细胞和毛囊 细胞和毛发，持续10天或更长时间(Li，L.，MarGolis， L.B.和霍夫曼，R.M.Proc.娜塔莉。阿卡德。科学，出版中)。它是 在这里提出，使用组织培养系统和荧光染料 测量细胞活性或细胞死亡，以调查一种 作为模型化合物的一系列农药、有机锡和致癌物质 研究各种组织培养小鼠皮肤的毒性和致癌作用 和人类包皮以及培养的皮肤上的毛发生长。这个 共聚焦极大地增强了对毒素影响的测量 显微镜可以对标本进行三维观察 这在以前是不可能的。至少一种化合物的体外作用 已经在体内使用裸鼠进行了相关研究，因此 体外的细胞毒性作用与刺激指数相对应 各种化合物在体内得分。

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.