YOUNG FROM STORED AND IMPAIRED MOUSE SPERM

来自储存和受损的小鼠精子的年轻小鼠

• 批准号: 6182469
• 负责人: JOHN D BIGGERS
• 金额: $ 46.21万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6182469
• 关键词: animal breeding; artificial fertilization; bioengineering /biomedical engineering; cryopreservation; embryo /fetus culture; embryo /fetus transplantation; genetic strain; genetically modified animals; in vitro fertilization; laboratory mouse; method development; sperm

The aim of this proposal is to develop an algorithm using assisted reproductive techniques to reconstitute valuable genetic strains of mice from stored spermatozoa. The ability to store spermatozoa is urgently needed because of the exponentially increasing numbers of mutant and transgenic strains of mice being produced for the study of human diseases. The cost of preserving these strains using conventional breeding programs is so astronomical that the storage of strains will soon be curtailed. The algorithm is also needed to facilitate the rescue of strains of mice with defective sperm or whose males are incapable of breeding. A working algorithm can be identified using current assisted reproductive techniques: artificial insemination, in vitro fertilization, intracellular sperm injection, embryo culture and embryo transfer. The overall efficiency of any particular path in this algorithm is defined as the number of pups born x 100/number of oocytes exposed to sperm. The overall efficiencies of the paths using IVF and ICSI, employing cryopreserved sperm and freeze-dried sperm spermatozoa from five strains of mice commonly used in bioengineering, will be determined. The results will reveal the extent that the overall efficiencies are genetically determined. The results will also allow a determination of whether the newly described freeze-drying route of preserving sperm is a workable option. Research is also proposed that will systematically examine the dehydration of spermatozoa from a biophysical point of view. Attempts will be made to optimize the freeze-drying techniques. Studies will also investigate whether the alternative evaporative route of dehydration is more effective. Other studies will be concerned with improving the current methods of in vitro fertilization, intracytoplasmic sperm injection and embryo transfer in mice, the results of which will be incorporated in a final algorithm.

该提案的目的是使用辅助生殖技术开发算法，以重新建立来自存储的精子的小鼠的有价值的遗传菌株。 由于用于研究人类疾病的研究的突变体和转基因菌株的数量呈指数增长，因此迫切需要储存精子的能力。 使用常规育种计划保存这些菌株的成本是如此天文学，以至于很快就会减少菌株的储存。还需要该算法来促进具有有缺陷的精子或雄性无法繁殖的小鼠菌株的营救。 可以使用当前辅助生殖技术来鉴定工作算法：人工授精，体外受精，细胞内精子注射，胚胎培养和胚胎转移。 该算法中任何特定路径的总体效率定义为出生的X 100/暴露于精子的卵母细胞数量。 使用IVF和ICSI的路径的总体效率，采用冷冻保存的精子和冻干的精子精子，来自通常用于生物工程的五种小鼠菌株。 结果将揭示总体确定总体效率的程度。 结果还将允许确定新描述的保留精子的冻干路线是可行的选择。还提出了研究从生物物理的角度系统地检查精子脱水的研究。将尝试优化冻干技术。研究还将研究替代的脱水途径是否更有效。 其他研究将涉及改善当前体外受精，胞质内精子注射和小鼠的胚胎转移的方法，其结果将纳入最终算法。