An Ultra-fast Cooling Device for Vitrification and Cryopreservation of Cells and Tissues

描述(由申请人提供)：将有价值的动物模式物种的基因类型作为活的动物品系进行存储和维护是完全不切实际的。然而，传统的冷冻保存技术很快就会被发现在为数百万正在开发的新的动物模型中的细胞和组织的冷冻保存提供经济的方法方面效率低下。有效的冷冻保存临床上有价值的组织，包括角膜组织、卵巢皮质条、皮肤、血管等，将对相应的移植技术和利用动物模型或人体组织进行基础生物医学研究具有重要意义。我们的总体目标是开发一种超快冷却设备(冷却速度106K/分钟)，以实现在没有或具有非常低浓度渗透性冷冻保护剂的情况下实现薄(约100-200微米)组织或样品的玻璃化。我们将通过应用包括低温薄膜蒸发和射流冲击冷却在内的尖端工程技术来生产超快速冷却设备的原型来实现这一点。为了提高其实际效率，该装置将被设计为一个封闭的操作系统，以防止样品直接接触液氮或其蒸汽。为实现这些目标，提出了第一阶段的计划：1)设计一种结合薄膜蒸发和射流冲击冷却技术的新型超快冷却装置，确定各子系统的最佳设计参数；2)制作该装置并测试其热效率；3)测试装置将用于猪和人角膜组织的冷冻保存，并将根据解冻后组织的生化和组织学测试来评估冷冻保存效率。在第一阶段结束时，建议的装置将在适当的工程设计控制下生产，并通过生物测试进行验证。

DESCRIPTION (provided by applicant): Storage and maintenance of valuable genotypes of animal model species as live animal lines would be wholly impractical. However, traditional cryopreservation techniques will be soon found to be inefficient for providing economic approach for cryopreservation of cells and tissues from millions of new animal models being developed. Efficient cryopreservation of clinically valuable tissues, including cornea tissues, ovarian cortical strips, skins, blood vessels, and many others will invaluably benefit corresponding transplantation technology and basic biomedical research using animal model or human tissues. Our overall goal is to develop an ultra-fast cooling device (cooling rate >106 K/min) to achieve vitrification of thin (approximately 100-200 microns) tissues or samples without or with very low concentration of permeating cryoprotectants. We will achieve this by producing the prototype of an ultra-fast cooling device with an application of cutting-edge engineering techniques including cryogenic thin film evaporation and jet impingement cooling. To improve its practical efficiency, the device will be designed as a close operating system that prevents samples from direct contact with liquid nitrogen or its vapor. To achieve those goals, the plan for phase I periods is proposed: 1) design a novel ultra-fast cooling device combining thin film evaporation and jet impingement cooling techniques, determine optimal design parameters for subsystems; 2) produce the device and test its thermal efficiency; and 3) the tested device will be used for porcine and human corneal tissue cryopreservation, and the cryopreserved efficiency will be assessed according to biochemical and histological tests on the post-thaw tissues. At the end of Phase I, the proposed device will be produced under appropriate engineering design control and validated through biological tests.