NOVEL SELF-NUCLEATING CRYOSTROGE DEVICES

描述（由申请人提供）：本发明涉及一种在低温保存（冷冻保存）期间在细胞和组织周围的水溶液中诱导冰形成（例如冰成核或“接种”）的方法。通常，通过用预冷仪器接触样品容器来进行接种。该过程使样品暴露于由于升温（如果将样品从冷却装置中取出以进行接种）或过度冷却和细胞内冰形成（如果细胞外冰的诱导将样品冷却得太快至非常低的温度）而导致的潜在损坏。该缺点通过提供化学诱导冰形成而不中断冷却过程或施加外部冷却的一般方法和特定装置来解决。该项目利用固醇化合物的冰成核特性，特别是胆固醇，结合到固态基质中自动诱导冰。本研究的总体目标涉及改进目前正在开发的这些新方法和装置，并测试它们在增加解冻后恢复和活力方面的有效性，特别是因为它涉及几种特定细胞类型的正常结构和功能的保留。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，检验固态成核系统将导致过冷减少，从而增加冻融人类精液的回收率的假设。 具体目标2。测试固态成核系统将导致过冷减少并因此增加冷冻解冻的脐带血造血干细胞的回收的假设。 具体目标3。使用标准的商业农业冷冻保存方法，检验固态成核系统将导致过冷减少并因此增加冷冻解冻公牛精液的回收率的假设。

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.