Rat sperm cryobanking for genetic resources

描述（由申请人提供）：种质冷冻库是一种科学、医学、经济和环境上重要的遗传资源无限期保存的有效方法。虽然大鼠是模拟人类疾病的重要系统，但目前还没有成熟的种质冷冻保存方案，可以对大鼠遗传资源进行经济有效的管理。收集附睾精子是最简单和最具成本效益的存档遗传学方法，以备将来使用。然而，到目前为止，只有一个方案显示只有10%的解冻后运动。因此，迫切需要了解影响大鼠精子的生理和低温生物学因素，以提高大鼠精子的低温存活率。本应用程序的目的是开发附睾大鼠精子的冷冻方案，该方案将允许使用标准的人工授精和体外受精方法重建遗传。在过去的几年中，我们的实验室已经产生了大量与最佳精子处理、Percoll分离、冷冻扩展剂以及大鼠精子的冷却和加热条件相关的数据。这将有助于减少物理、低温、渗透压力和致命的冰形成造成的伤害，并最终产生可接受的解冻后恢复速率。我们在实验室收集的数据，结合本申请中概述的拟议的冷冻生物学研究，将使我们能够开发有效和可靠的大鼠附睾精子冷冻保存方案。

DESCRIPTION (provided by applicant): Germplasm cryobanking is an effective way of preserving scientifically, medically, economically and environmentally important genetic stocks indefinitely. Although rats are important system for modeling human disease, there is no established germplasm cryopreservation protocol which would allow cost-effective management of rat genetic resources. Collection of epididymal sperm is the simplest and most cost-effective method of archiving genetics for future use. However, to date, there is only one protocol showing only 10% post-thaw motility. Thus, there is an urgent need for understanding the physiological and cryobiological factors affecting rat sperm in order to improve the rat sperm cryosurvival. The goal of this application is to develop freezing protocols for epididymal rat sperm which would allow reconstitution of genetics by using standard artificial insemination and in-vitro fertilization methods. Over the last few years, our laboratory has generated ample amount of data related with optimal sperm handling, Percoll separation, freezing extender and cooling and warming conditions for the rat sperm. These would help reduce the injuries caused by physical, hypothermic, osmotic stress and lethal ice formation, and ultimately yield acceptable post-thaw recovery rate. The data which have been collected in our laboratory in combination with proposed cryobiologic studies outlined in this application will allow us to develop efficient and reliable cryopreservation protocol for epididymal rat sperm. PUBLIC HEALTH RELEVANCE (provided by applicant): This proposal aims at developing efficient cryopreservation method for rat sperm which would be used to reconstitute genetically important rat strains through commonly used assisted reproductive technologies such as artificial insemination or in-vitro fertilization if needed. The proposed studies will therefore address to the biomedical community by providing a reliable protocol for rat sperm cryopreservation in order to avoid continuous live animal housing which is a significant burden for National Institutes of Health and its investigators around the nation.