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

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了解提供了益处，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 发起了大鼠基因组计划和大鼠基因组数据库。美国国立卫生研究院主办的大鼠模型优先事项会议建议建立国家大鼠遗传资源中心，支持大鼠种系修饰的研究，包括为大鼠精子冷冻保存研究提供专项资金，以缓解储存和运输问题，并改进大鼠体外受精技术。不幸的是，目前大鼠精子冷冻保存的技术水平很差。对文献的电子检索显示，过去 40 年中仅发表了两篇旨在优化大鼠精子冷冻保存的论文（Nakatsukasa 等人，2001 年；2003 年）。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、了解冷冻保存对精子造成的损害或描述该物种的冷冻保存过程。在本提案中，我们将尝试实现三个非常重要的目标，因为它涉及一般精子冷冻保存，特别是大鼠精子冷冻保存。首先，我们将尝试通过优化冷冻速率来优化大鼠精子冷冻保存，该冷冻速率基于冷冻过程中精子水传输特性的仔细表征和冷冻保护介质的优化。其次，我们将在分子和结构水平上确定冷冻保存过程对精子造成的损害，并根据我们对机制的理解设计和测试方法以尽量减少这种损害。最后，我们将标准化方法以减少伤害并优化冷冻保存结果。这些方法体现在以下 3 个具体目标中： 具体目标 1：确定对大鼠精子冷冻保存最有效的处理方法和 CPA 介质。 具体目标 2：确定冷冻过程中大鼠精子质膜的生物物理水运输特性。 具体目标 3：设计和测试保护大鼠精子免受冷冻保存损伤的策略。

项目成果

Cellular biophysics during freezing of rat and mouse sperm predicts post-thaw motility.

大鼠和小鼠精子冷冻过程中的细胞生物物理学预测解冻后的活力。

• 发表时间: 2009-10
• 影响因子: 3.6
• 作者: Hagiwara, Mie;Choi, Jeung Hwan;Devireddy, Ramachandra V;Roberts, Kenneth P;Wolkers, Willem F;Makhlouf, Antoine;Bischof, John C
• 通讯作者: Bischof, John C