Cryopreservation of a Living Human Skin Equivalent

描述(申请人提供)：生产活体人体皮肤 相当于(LSE)，经济、机械耐用，并提供 治疗上皮伤(如烧伤)的有效屏障功能 和糖尿病病变)是当前组织研究的一个非常有价值的目标 工程学。Stratech，L.L.C.目前正在将各种产品商业化 基于专利的人类角质形成细胞系NIKStm，已经被 显示为完全分层的活体皮肤等价物 组织学标志物表达正常。保质期有限，保质期高 目前可用的皮肤等效性产品的存储成本限制了它们的 治疗潜力。我们假设一个无毒的冷冻防护剂系统 基于专门设计用于在冷却时形成玻璃的糖类(即， 玻璃化)比冷冻更能有效地保存皮肤 组织结构和活性比目前的方法(即，治疗 甘油)。高效超低温保存技术的研究进展 NIKStm细胞系代表着为医学界提供 供应急需的现成皮肤移植材料和长货架 在相对较高的储存温度下生活。 建议的商业应用： 随着组织工程领域的发展，对未来产品的有效、无毒保存方法的需求越来越大。Stratech正在开发基于我们专有的NIKS(TM)细胞系的活体人类皮肤仿制品。这项提议的目标是在冷冻保存的最新进展的基础上，结合将保护剂引入靶细胞的方法来开发技术。我们推测，通过增强玻璃的形成，可以实现一种无毒、高储存温度的活体人体皮肤替代物保存系统，这是一种具有很高治疗价值的模型系统。我们相信，这项技术将适用于广泛的其他组织和器官。

DESCRIPTION (provided by applicant): Production of a living human skin equivalent (LSE) that is economical, mechanically durable and provides effective barrier function for the treatment of epithelial trauma (e.g., burns and diabetic lesions) is a highly prized goal of current efforts in tissue engineering. Stratatech, L.L.C., is currently commercializing various products based on the proprietary human keratinocyte cell line, NIKStm, which has been shown to differentiate into a fully stratified living skin equivalent with normal expression of histological markers. The limited shelf life and high storage costs for currently available skin equivalent products limit their therapeutic potential. We hypothesize that a non-toxic cryoprotectant system based on saccharides specifically designed to form a glass on cooling (i.e., vitrify) rather than freeze will be more effective in the preservation of skin tissue architecture and viability than current methods (i.e., treatment with glycerol). Development of highly effective cryopreservation methods for the NIKStm cell line represents an opportunity to provide the medical community with a much needed supply of off-the-shelf skin graft material with long shelf life at relatively high storage temperatures. PROPOSED COMMERCIAL APPLICATION: As the field of tissue engineering advances, the need for effective, non-toxic preservation methods for future products grows. Stratatech is developing living human skin equivalent products based on our proprietary NIKS (TM) cell line. The goal of this proposal is to develop technology based on recent advances in cryopreservation coupled with methods to introduce protectants into target cells. We hypothesize that by enhancing glass formation a non-toxic, high storage temperature preservation system can be achieved for living human skin equivalents, a model system with high therapeutic value. It is our belief that this technology will be applicable to a wide range of other tissues and organs.