Hypothermic Storage and Cryopreservation of Corneas

描述(申请人提供)：细胞保存服务公司(CPSI)是一家生物技术公司，致力于低温生物和医学领域的新技术开发。CPSI的核心分子策略专注于理解和操纵细胞生存和细胞死亡途径，这些途径似乎是细胞特有的，并因低温暴露而被激活。这一范式转变导致了各种人类细胞和组织的低温储存和冷冻保存过程的改进。最近，通过初步研究，我们已经确定，仅仅通过溶液设计来解决保存过程中的应激反应问题是不够的，这是过去的重点，但实际上需要一种更全面、更综合的策略，包括保存前和保存后的细胞调节策略。CPSI应用了这一新的战略方法，并确定了似乎可以改善角膜细胞和组织质量的有前途的新方法，而不是仅依赖于使用Optisol-GS等保存介质来维持组织完整性和功能的典型方案。在人角膜内皮细胞(HCEC)、牛角膜内皮细胞(BCEC)和其他相关的人类细胞株上收集的可行性数据表明，开发改进的角膜加工和保存方案是高度可行的，成功地开发一种一体化的过程不仅应该提高角膜组织的质量和用途，而且还应该增加供体来源。在这项研究项目下进行的研究将利用cDNA微阵列、蛋白质组图谱以及活性和功能分析来确定在移植前处理中常见的低温暴露后在HCEC中激活的应激反应通路。这些研究的目的是为了更好地了解HCEC和整个角膜对低温暴露的独特反应，并利用这些数据开发新的角膜组织处理程序。我们已经与哈佛医学院、谢彭斯眼科研究所、莫兰眼科中心和NDRI等多个组织的研究人员建立了合作和分包合同，以促进各种外部资源的获取(包括专业知识、细胞培养系统、知识产权等)。评估在该研究项目下开发的新流程/协议，并帮助确保该研究计划的成功。我们相信，拟议的调查将导致处理角膜组织的新方法。这些进展不仅应该对收获的角膜的质量和可用性产生影响，而且还将使角膜来源细胞在体外毒理学测试和HCEC移植等领域得到更广泛的利用。角膜混浊是导致失明的主要原因。解决这种视力障碍的一种方法是将供体角膜移植给患者。这样做通常意味着供体角膜在移植前必须在寒冷的暂停存活状态下保存。过去使用Optisol-GS开发的方法已经过时，没有基于现代分子医学。CPSI建议使用一种分子策略来开发改进的人体角膜长期存储方案-一套集成的过程，既可以增加可用的角膜捐赠池，也可以提高角膜移植市场的组织质量。

DESCRIPTION (provided by applicant): Cell Preservation Services, Inc. (CPSI) is a biotechnology company dedicated to the development of novel technologies in the area of low temperature biology and medicine. CPSI's core molecular strategy is focused on understanding and manipulating the cell survival and cell death pathways that appear to be cell-specific and are activated as a consequence of low temperature exposure. This paradigm shift has led to improved processes for the hypothermic storage and cryopreservation of a variety of human cells and tissues. Recently, through preliminary studies, we have determined that it is not adequate to merely address stress response issues during preservation through solution design, as has been the focus in the past, but in fact a more comprehensive and integrated strategy involving pre- and post-preservation cell conditioning strategies is necessary. CPSI applied this novel strategic approach and has identified promising new methodologies that appear to improve corneal cell and tissue quality beyond that of typical protocols which rely only on the use of preservation media, such as Optisol-GS, to maintain tissue integrity and function. Feasibility data gathered on human corneal endothelial cells (HCEC), bovine corneal endothelial cells (BCEC) and other relevant human cell strains suggest that the development of improved cornea processing and preservation protocols is highly feasible and successful development of an integrated process should not only increase the quality and utility of corneal tissue, but increase the donor pool as well. Investigations conducted under this research project will utilize cDNA microarrays, proteomic profiling, and viability and functional analysis to define the stress response pathways activated in HCEC following low temperature exposure common to pre-transplant processing. It is the intent of these studies to provide a better understanding of the unique responses of HCEC and whole cornea to low temperature exposure and utilize these data to develop new procedures for corneal tissue processing. We have established collaborations and subcontracts with researchers from several organizations including Harvard Medical School, the Schepens Eye Research Institute, the Moran Eye Center and NDRI to facilitate access to various outside resources (including expertise, cell culture systems, IP, etc.) to evaluate new processes/protocols that are developed under this research project as well as to help ensure the success of this research program. It is our belief that the proposed investigations will lead to new methodologies for the processing of corneal tissue. These developments should have an impact not only on the quality and availability of harvested corneas, but also enable expanded utilization of corneal-derived cells in areas such as in vitro toxicological testing and HCEC transplant. Corneal opacities are leading causes of blindness. One method to resolve this impediment to clear vision is to transplant donor corneas to the patient. To do so typically means that donor corneas must be preserved in a state of suspended animation in the cold prior to transplant. The methodologies developed in the past using Optisol- GS are dated and not based on modern, molecular medicine. CPSI proposes to use a molecular strategy to develop improved protocols for the long term storage of human corneas - a set of integrated processes that will both increase the available cornea donor pool as well as the quality of the tissue for the cornea transplant market.