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

描述（由申请方提供）：将动物模型种属的有价值基因型作为活动物系储存和维持是完全不切实际的。然而，传统的冷冻保存技术将很快被发现是低效的，为数以百万计的新动物模型的细胞和组织的冷冻保存提供经济的方法。角膜组织、卵巢皮质条、皮肤、血管等具有临床价值的组织的有效冷冻保存将为相应的移植技术和动物模型或人体组织的基础生物医学研究带来不可估量的价值。我们的总体目标是开发一种超快速冷却装置（冷却速率>106 K/min），以在没有或具有非常低浓度的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。我们将通过生产超快速冷却装置的原型来实现这一目标，该装置应用了尖端工程技术，包括低温薄膜蒸发和射流冲击冷却。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，防止样品与液氮或其蒸气直接接触。为实现上述目标，提出了第一阶段的研究方案：1）设计一种新型的薄膜蒸发与射流冲击冷却相结合的超快速冷却装置，确定各子系统的最佳设计参数; 2）制作装置并测试其热效率;以及3）受试装置将用于猪和人角膜组织的冷冻保存，并根据解冻后组织的生化和组织学试验评估冷冻保存效率。在第一阶段结束时，将在适当的工程设计控制下生产申报器械，并通过生物学试验进行确认。

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.