DEVELOPMENT OF AN EMBRYO CRYOPRESERVATION PROGRAM IN LABORATORY ANIMALS

该计划的目标是促进NLH的管理 动物遗传资源(NlHAGR)的开发和利用 利用采集、培养、冷冻等技术 以及胚胎和配子的移植用于冷藏库基因 材料和提高生殖潜力。NlHAGR 维持着一些关键物种，在物种内部， 在生物医学研究中使用的宝贵的基因类型的数量。胚胎 在冷冻介质中使用冷冻保护剂进行保存 允许充分脱水，以保护卵裂球在 冷得要命。在冷冻保护剂解冻和稀释后， 胚胎被培养以评估体外发育。 将胚胎移植到受体雌性体内的监视器 发展，因此生物能力。体外受精 和卵子冷冻作为挽救卵子的技术被研究 来自单身雌性或贫穷品系的有限遗传物质 体内受精率。将储存的胚胎保存在 冻结状态，直到需要重新派生。的主要关注点 实验室的努力是研究和优化低温库 技术在老鼠身上，最近在兔子身上。一项重大研究 研究发现，胚胎的冷冻能力受到了混淆和影响 明显受小鼠基因的影响。这样的回应已经在 超过55种基因类型，其中大多数显示出特定于基因的存活率 标准化胚胎冷冻解冻程序的比率。辅助的 项目着眼于改进的比较方面 胚胎的长期保存，特别强调对 胚胎冷冻、解冻、冷冻的保护剂及强化方法 稀释和转移。兔子的努力集中在改进上 在胚胎采集和保存过程中使用一种新的 开发了玻璃化冷冻程序。

The program objective is to facilitate the management of the NlH Animal Genetic Resource (NlHAGR) through development and utilization of techniques such as collection, culture, freezing and transfer of embryos and gametes for cryobanking genetic material and improving reproductive potential. The NlHAGR maintains a number of critical species and, within species, vast number of invaluable genotypes used in biomedical research. Embryo preservation uses a cryoprotectant in the freezing medium which allows sufficient dehydration to protect the blastomere during freezing. Following thawing and dilution of the cryoprotectant, embryos are cultured to assess in vitro development. Transplantation of embryos into recipient females monitors in vivo development and thus biological competence. In vitro fertilization and ova freezing are being examined as techniques to salvage limited genetic material from single females or strains with poor in vivo fertilization rates. Banked embryos are maintained in a frozen state until needed for rederivation. A primary focus of laboratory efforts is the study and optimization of cryobanking technology in mice and more recently, in rabbits. A major research finding is that embryo freezability is confounded and influenced markedly by mouse genotype. Such responses have been examined in over 55 genotypes, most of which display genotype specific survival rates to standardized embryo freeze thawing procedures. Ancillary projects are oriented toward the comparative aspects of improving long-term embryo storage with particular emphasis on the study of cryoprotectants and enhancing methods for embryo freezing, thawing, dilution and transfer. Efforts in the rabbit focus on improvements in collection procedures and preservation of embryos using a newly developed vitrification procedure.