NOVEL SELF-NUCLEATING CRYOSTROGE DEVICES

描述（由申请人提供）：本提案涉及在低温保存（冷冻保存）期间在细胞和组织周围的水溶液中诱导冰形成（例如冰核或“播种”）的方法。通常，播种是通过用预冷仪器触摸样品容器来进行的。由于升温（如果样品从冷却装置中取出进行播种）或如果细胞外冰的诱导使样品过快地冷却到非常低的温度，则广泛的冷却和细胞内冰的形成，该过程使样品暴露于潜在的损坏中。通过提供一种通用的方法和特定的装置，在不中断冷却过程或应用外部冷却的情况下化学诱导冰的形成，解决了这一缺点。该项目利用甾醇化合物，特别是胆固醇的冰核特性，结合到固态基质上自动诱导冰。本研究的总体目标是完善这些目前正在开发的新方法和设备，并测试它们在提高解冻后恢复和生存能力方面的有效性，特别是在保留几种特定细胞类型的正常结构和功能方面。为实现这一目标，将执行以下具体目标：

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.