NOVEL SELF-NUCLEATING CRYOSTROGE DEVICES

描述(由申请人提供)：本提案涉及一种在低温保存(低温保存)期间在细胞和组织周围的水溶液中诱导结冰(例如，冰核或“播种”)的方法。通常，播种是通过用预冷仪器接触样品容器来执行的。这一过程使样品暴露在升温(如果样品被从冷却设备中取出进行播种)或广泛冷却和细胞内冰形成的潜在损害中，如果细胞外冰的诱导使样品过快冷却到非常低的温度。通过提供一种在不中断冷却过程或应用外部冷却的情况下化学诱导结冰的通用方法和特定装置来解决这一缺点。该项目利用与固态基质结合的甾醇化合物的成冰特性，特别是胆固醇，自动诱导结冰。这项研究的总体目标是完善目前正在开发的这些新方法和设备，并测试它们在提高解冻后恢复和活性方面的有效性，特别是在与保留几种特定细胞类型的正常结构和功能有关的情况下。为实现这一目标，将实现以下具体目标： 具体目标1.使用标准的商业精子库方法，测试固态核系统将减少过冷，从而提高冷冻解冻的人类精液回收率的假设。 具体目的2.验证这样的假设：固态成核系统将减少过冷，从而提高冷冻解冻的脐带血造血干细胞的回收率。 具体目标3.使用标准的商业农业冷冻保存方法，测试固态核系统将减少过冷，从而提高冷冻解冻公牛精液回收率的假设。

DESCRIPTION (provided by applicant): The present proposal relates to a method of inducing ice formation (e.g. ice nucleation or "seeding") in aqueous solutions surrounding cells and tissues during low temperature preservation (cryopreservation). Typically, seeding is performed by touching the sample containers with a precooled instrument. This process exposes the samples to potential damage due to warming (if the samples are removed from the cooling device to perform seeding) or extensive cooling and intracellular ice formation if the induction of extracellular ice cools the sample too quickly to very tow temperatures. This shortcoming is addressed by providing a general method and specific devices that chemically induce ice formation without interruption of the cooling process or application of external cooling. This project utilizes the ice nucleating properties of sterol compounds, specifically cholesterol, bound to solid state matrices to automatically induce ice. The overall goal of this research relates to refining these novel methods and devices currently under development, and test the effectiveness of them to increase post-thaw recovery and viability, especially as it relates to retention of normal structure and function, of several specific cell types. To achieve this goal, the following specific aims will be performed: Specific Aim 1. Test the hypothesis that the solid-state nucleating system will result in a reduction of supercooling and therefore an increased recovery of frozen-thawed human semen using standard, commercial sperm banking methods. Specific Aim 2. Test the hypothesis that the solid-state nucleating system will result in a reduction of supercooling and therefore an increased recovery of frozen thawed umbilical cord blood hematopoietic stem cells. Specific Aim 3. Test the hypothesis that the solid-state nucleating system will result in a reduction of supercooling and therefore an increased recovery of frozen thawed bull semen using standard commercial agriculture methods of cryopreservation.