EFFICACY OF MOUSE SPERM CRYOPRESERVATION

小鼠精子冷冻保存的功效

• 批准号: 6018012
• 负责人: LARRY E MOBRAATEN
• 金额: $ 49.71万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-20 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6018012
• 关键词: cryopreservation; fertility; genetic strain; germ free condition; in vitro fertilization; laboratory mouse; sperm; spermatogenesis

DESCRIPTION: (Adapted from Applicant?s Description) Revolutionary advancements in genome research and the ability to create genetically specific strains of mice are resulting in an exponential increase in the number of strains available for biomedical research. Freezing and storing sperm could most efficiently preserve the majority of these newly created strains, but existing methods do not work sufficiently well for all strains of mice. The long-term objective of this project is to improve methods of sperm cryopreservation for reliable recovery of pathogen free and reproductively normal offspring. The specific aims of this project are to: (1) Establish efficient freezing protocols for improved cryopreservation of sperm from different inbred strains. Various additions and substitutions of components in an existing cryoprotectant formulation will be made within a narrow range of osmolality and tested for their effect on the efficiency of successfully freezing inbred and mutant strain sperm. (2) Enhance in vitro fertilization (IVF) for gametes from different inbred strains. Various modifications of IVF such as addition of free radical scavengers to the culture medium and removal of the zona pellucida from oocytes will be tested for ability to enhance fertilization of inbred strain gametes. (3) Determine the risk of pathogen transmission through cryopreserved sperm upon recovery of offspring through IVF or intracytoplasmic sperm injection (ICSI) using sperm of infected males sent to The Jackson Laboratory for re-derivation and importation of a strain that will be diagnosed for presence of pathogens. (4) Provide quantitative assessment of reproductive function in the animals produced from frozen sperm. Females and males produced from cryopreserved sperm will be mated to ICR mice to evaluate their fertility and will be tested for reproductive behaviors, measured for plasma levels and in vitro release of reproduction-related hormones, and assessed for spermatogenesis. Neuroendocrine mechanisms that may underlie any reproductive deficits found will be identified.

描述：（改编自申请人的描述）革命进步 在基因组研究中以及创建遗传特定菌株的能力 小鼠导致菌株数量呈指数增加 可用于生物医学研究。冻结和储存精子最可能 有效地保留这些新创建的菌株的大多数，但现有 对于所有小鼠菌株，方法不能很好地工作。长期 该项目的目的是改善精子冷冻保存方法 可靠的无病原体恢复和生殖正常后代。这 该项目的具体目的是： （1）建立有效的冻结协议，以改善对的冷冻保存 来自不同近交菌株的精子。各种补充和替换 现有的冷冻保护剂配方中的组件将在 渗透范围狭窄，并测试了它们对效率的影响 成功冻结近交和突变菌株精子。 （2）增强来自不同近交的配子的体外受精（IVF） 菌株。 IVF的各种修改，例如添加自由基 清除培养基，并从 将测试卵母细胞的能力，以增强近交菌株的受精 配子。 （3）确定通过冷冻保存精子传播病原体的风险 通过IVF或室内精子注射恢复后代后 （ICSI）使用被送往杰克逊实验室的被感染男性重新衍生的精子 并导入将诊断为存在的菌株 病原体。 （4）对动物的生殖功能进行定量评估 由冷冻精子产生。由冷冻保存产生的女性和男性 精子将与ICR小鼠配对以评估其生育能力，并将进行测试 对于生殖行为，测量了血浆水平和体外释放 繁殖相关的激素，并评估了精子发生。 可能是任何生殖缺陷的基础的神经内分泌机制 将被确定。