ORGANOTYPIC SKIN MODELS CONTAINING MELANOCYTES

第一阶段的研究表明，正常的人类黑素细胞可以 直接添加到高度分化的无血清组织培养物中 人体表皮的模型。 其中的黑色素细胞 保留了它们的高度树枝状的性质，并被定位在 组织培养构建体的基底细胞层。 底物I-多巴 转化为黑色素，超微结构分析显示， 黑素细胞内含有大量黑素体， 其中一些转移到相邻的角质形成细胞。 最后， 模型的初步特征表明，黑素细胞确实 不会不利地影响本发明的任何有利性质。 表皮模型 第二阶段的研究将建立在第一阶段的结果，以进一步发展 这个三维的含有黑素细胞的组织模型 准确、有用和商业上可行的产品。 初步研究将 探索潜在的营养或激素缺乏， 存在于培养基中。 黑素细胞的选择和 其在培养物中的处理也将被优化。 进一步 模型的特性，特别是那些将 受影响的是暴露于紫外线照射（UV），沿着 模型的重现性，将进行研究。 此外， 该模型将使用对黑素生成的外源刺激来探测。 最后，使用从白癜风皮肤中获得的黑素细胞， 代表了这个相对常见的 皮肤病 据信，这种组织结构将有助于 阐明一些白癜风的基础，目前还没有 明白 建议的商业应用 这项拟议中的研究将导致商业上可行的黑素细胞- 包含皮肤模型，将帮助公司开发 影响皮肤色素沉着或受其影响的产品。 皮肤 模型将在理解黑素生成和调节剂方面找到实用性 研究皮肤色素沉着疾病，并阐明 紫外线对表皮的损伤机制。 此外，本发明还提供了一种方法， 白癜风组织构建体将帮助皮肤科公司 开发药物和/或疗法来治疗这种广泛的 皮肤色素沉着病。

Phase I research demonstrated that normal human melanocytes could be directly added to a highly differentiated, serum-free tissue culture model of the human epidermis. The melanocytes incorporated therein retained their highly dendritic nature and were localized within the basal cell layer of the tissue culture construct. The substrate, I-dopa, was converted to melanin and ultrastructural analysis revealed the presence of numerous melanosomes contained within the melanocytes, some of which were transferred to adjacent keratinocytes. Finally, initial characterization of the model showed that the melanocytes did not deleteriously affect any of the advantageous properties of the epidermal model. Phase II research will build on the Phase I results to further develop the three-dimensional, melanocyte-containing tissue models into accurate, useful, and commercially viable products. Initial studies will probe potential nutritional or hormonal deficiencies which may be present in the culture medium. The choice of melanocytes and the handling thereof in culture will also be optimized. Further characterization of the model~s properties, particularly those which will be affected be exposure to ultraviolet irradiation (UV), along with the model~s reproducibility, will be studied. In addition, the accuracy of the model will be probed using exogenous stimuli to melanogenesis. Finally, the use of melanocytes obtained from vitiliginous skin will represent the first three-dimensional model of this relatively common skin disorder. It is believed that such tissue constructs will help elucidate some of vitiligo~s underpinnings which currently are not understood. PROPOSED COMMERCIAL APPLICATION The proposed research will lead to commercially viable, melanocyte- containing skin models which will assist companies in developing products which affect, or are affected by, skin pigmentation. The skin models will find utility in understanding melanogenesis and modulators thereof, studying skin pigmentation disorders, and elucidating mechanisms of UV light induced damage to the epidermis. In addition, the vitiligo tissue constructs will assist dermotologic companies in developing pharmaceuticals and /or therapies to treat to this widespread skin pigmentation disease.