New Approaches to Rat Sperm Cryopreservation

大鼠精子冷冻保存的新方法

• 批准号: 7112299
• 负责人: JOHN C BISCHOF
• 金额: $ 17.58万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-15 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7112299
• 关键词: New; Approaches; Rat; Sperm; Cryopreservation; New; Approaches; Rat; Sperm; Cryopreservation

DESCRIPTION (provided by applicant): In light of the benefit that the rat model system have provided to our basic understanding of the biology of human disease, and the importance of the rat to emerging fields of genomics and stem cell biology, the NIH initiated the Rat Genome Program and the Rat Genome Database. The NIH sponsored meeting on Rat Model Priorities recommended that a National Rat Genetic Resource Center be established to support research on germ-line modifications in the rat, including ear-marked funding for the study of rat sperm cryopreservation to alleviate the storage and transportation problems, and to improve in vitro fertilization techniques in the rat. Unfortunately, the current state of the art in rat sperm cryopreservation is abysmal. An electronic search of the literature reveals only two papers published in the last 40 years that seeks to optimize the cryopreservation of rat sperm (Nakatsukasa et al. 2001; 2003). This work establishes the feasibility of rat sperm cryopreservation but makes no attempt to optimize the technology, understand the damage caused to the sperm by cryopreservation, or to characterize the cryopreservation process in this species. In this proposal, we will attempt to achieve three very important goals as it relates to sperm cryopreservation in general and rat sperm cryopreservation in particular. First, we will attempt to optimize rat sperm cryopreservation by optimizing the freezing rate based careful characterization of the water transport properties of the sperm during freezing and optimization of the cryoprotective media. Second, we will determine the damage done to the sperm, at the molecular and structural level, caused by cryopreservation process, and design and test methods to minimize this damage based on our understanding of the mechanism. Finally, we will standardize approaches to reduce injury and optimize cryopreservation outcomes. These approaches are reflected in the following 3 specific aims: Specific Aim 1: Determine the handling methods and CPA media most effective for rat sperm cryopreservation. Specific Aim 2: Determine the biophysical water transport properties of rat sperm plasma membrane during freezing. Specific Aim 3: Design and test strategies for protecting rat sperm from damage incurred by cryopreservation.

描述（由申请人提供）：鉴于大鼠模型系统为我们对人类疾病生物学的基本理解提供的益处，以及大鼠对基因组学和干细胞生物学新兴领域的重要性，NIH启动了大鼠基因组计划和大鼠基因组数据库。 NIH赞助的大鼠模型优先级会议建议建立一个全国大鼠遗传资源中心，以支持对大鼠种系修饰的研究，包括用于研究大鼠精子冷冻保存研究以减轻储存和运输问题的资金，并改善大鼠的体外受精技术。不幸的是，大鼠精子冷冻保存中的最新水平是糟糕的。对文献的电子搜索揭示了过去40年中仅发表的两篇论文，旨在优化大鼠精子的冷冻保存（Nakatsukasa等，2001; 2003）。这项工作确立了大鼠精子冷冻保存的可行性，但没有尝试优化技术，了解通过冷冻保存对精子造成的损害，或者是该物种中的冷冻保存过程对精子造成的。在此提案中，我们将尝试实现三个非常重要的目标，因为它尤其是与精子冷冻保存有关，尤其是大鼠精子冷冻保存。首先，我们将尝试通过在冷冻和优化冷冻保护介质期间优化基于冰冻速率的冻结速率来优化大鼠精子冷冻保存。其次，我们将在分子和结构水平上确定由冷冻保存过程以及设计和测试方法在分子和结构水平上造成的损害，以根据我们对机制的理解来最大程度地减少这种损害。最后，我们将标准化减少伤害和优化冷冻保存结果的方法。这些方法反映在以下3个特定目的中：特定目的1：确定处理方法和CPA介质最有效地对大鼠精子冷冻保存。 特定目标2：确定冰冻过程中大鼠精子质膜的生物物理水传输特性。 特定目的3：设计和测试策略，以保护大鼠精子免受冷冻保存造成的损害。