ORGANOTYPIC SKIN MODEL CONTAINING MELANOCYTES

许多在体外高度分化的表皮和真皮/表皮样蛋白 已经开发了用于以下方面的模型：1)透皮药物 给药系统，2)皮肤移植，3)皮肤刺激性 测试。在很大程度上，这些模型包含了表皮 角质形成细胞和/或真皮成纤维细胞但不包括黑素细胞， 珠被的色素细胞提供保护，使其免受 潜在有害的紫外线(UV)太阳辐射。此外, 黑素细胞被认为参与了 皮肤，显然需要允许研究皮肤色素沉着 疾病、紫外线引起的皮肤损伤等。 利用无血清、高分化角质形成细胞的表皮 以之前开发的模型为起点，本研究将尝试 开发类似的无血清培养系统，使 黑素细胞的掺入。将开发一种无血清培养液，使用 已知的可刺激细胞增殖和维持的添加剂 黑素细胞的单层培养。三维，高度 分化的表皮结构将被培养和鉴定 使用光学和电子显微镜，黑素细胞生化标记物，以及 黑色素合成。最后，对该模型进行了初步探讨。 对已知影响体内黑素细胞的刺激的反应，如S紫外线将 制造。这些研究将为第二阶段研究奠定基础 将包括深入分析紫外线对表皮的影响， 皮肤的光老化，以及其他与黑素细胞相关的皮肤现象。

Many highly differentiated in vitro epidermal and dermal/epidermal sin models have been developed which are useful for: 1) transdermal drug delivery systems, 2) skin transplantation, and 3) dermal irritancy testing. For the most part, these models incorporate epidermal keratinocytes and/or dermal fibroblasts but do not include melanocytes, the pigment cells of the integument which provide protection from potentially harmful ultraviolet (UV) solar irradiation. In addition, melanocytes are believed to participate in the inflammatory response of skin and are obviously required to allow the study of skin pigmentation disorders, UV induced skin damage, etc. Utilizing a serum free, highly differentiated keratinocyte based epidermal model previously developed as a starting point, this research will attempt to develop a similar serum free culture system which allows the incorporation of melanocytes. A serum free medium will be developed using additives known to stimulate the proliferation and maintenance of melanocytes in monolayer culture. Three dimensional, highly differentiated epidermal constructs will be cultured and characterized using light and electron microscopy, a melanocyte biochemical marker, and melanin synthesis. Finally, an initial investigation of the model's response to stimuli known to affect melanocytes in vivo such s UV will be made. These studies will lay the groundwork for Phase II research which will include an in-depth analysis of the effect of UV on the epidermis, photoaging of skin, and other melanocyte related skin phenomena.