ORGANOTYPIC SKIN MODELS CONTAINING MELANOCYTES

第一阶段的研究表明，正常人的黑素细胞可以 直接加入高度分化的无血清组织培养中 人类表皮的模型。其中包含的黑素细胞 保留了它们高度树枝状的性质，并定位于 组织培养构建体的基本细胞层。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小体， 其中一些被转移到邻近的角质形成细胞。最后， 对该模型的初步描述表明，黑素细胞 不会有害地影响 表皮模型。 第二阶段研究将在第一阶段成果的基础上进一步发展 将含有黑素细胞的三维组织模型 准确、有用和商业上可行的产品。初步研究将 探索潜在的营养或荷尔蒙缺乏，可能是 存在于培养基中。黑素细胞的选择和 它在培养中的处理也将得到优化。进一步 模型的表征~S的性质，特别是那些将 受到紫外线照射(UV)的影响，以及 模型~S的重现性，将进行研究。此外，该方法的准确性 该模型将使用外源性刺激黑素生成进行探索。 最后，使用从白斑皮肤获得的黑素细胞将 代表了这一相对常见的第一个三维模型 皮肤病。人们相信，这种组织结构将有助于 阐明白癜风的一些基础~S目前还没有 明白了。 建议的商业应用 这项拟议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 影响或受皮肤色素沉着影响的产品。皮肤 模型将在理解黑素生成和调节剂方面找到有用之处 其中，研究皮肤色素沉着障碍，并阐明 紫外光诱导表皮损伤的机制。此外, 白癜风组织结构将帮助皮肤病公司 开发药物和/或疗法来治疗这种广泛传播的疾病 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.