腹腹肿胀作为辅助生殖用B6C3F1受体小鼠妊娠成功的指标。

Christina Gougoula, W Peter M Benten, Ani Kaplanian, Laurentiu Benga, Jeanette Knorr, Eva Engelhardt, Martin Sager
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摘要

体外受精(IVF)、胚胎冷冻保存和胚胎移植(ET)是辅助生殖技术(ARTs),在动物研究中广泛用于维持小鼠模型。不同遗传背景的近交系小鼠在生殖性能上存在差异。低温保存会影响胚胎的质量和活力,而体外受精接受者的遗传背景也会影响体外受精的结果。在这项回顾性研究中,我们分析了过去6年中在我们的设施中进行的et的结果。我们发现,无论是新鲜分离的胚胎还是冷冻解冻的胚胎植入,壶腹肿胀的B6C3F1小鼠的妊娠率都比壶腹未肿胀的小鼠高出近3倍。将新鲜收集的胚胎植入壶腹肿胀的受者体内,无论冷冻解冻胚胎是体内受精还是体外受精,其妊娠率都明显高于冷冻解冻胚胎。此外,我们发现遗传背景对出生率有显著影响;C57BL/6J小鼠和混合遗传背景小鼠的出生率比C57BL/6N小鼠高34%。在C57BL/6J组中,使用新鲜的体内受精胚胎时,出生率显著提高,低温保存对体内和体外受精胚胎均有负面影响。冻融胚胎与新鲜胚胎获得1只幼崽的成功率无显著差异。总的来说,壶腹肿胀是成功怀孕的一个强有力的指标,再加上胚胎操作和遗传背景。更好地了解影响生殖结果的因素可能会导致ART方案的优化,并有助于减少用于这些程序的小鼠数量。
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Swollen Ampulla as an Indicator of Successful Pregnancy in B6C3F1 Recipient Mice used for Assisted Reproduction.

In vitro fertilization (IVF), embryo cryopreservation, and embryo transfer (ET) are assisted reproductive technologies (ARTs) that are used extensively for the maintenance of mouse models in animal research. Inbred mouse strains with different genetic backgrounds vary in their reproductive performance. Cryopreservation can affect embryo quality and viability, and the genetic background of ET recipients can influence the ET result. In this retrospective study, we analyzed the out- comes of ETs performed in our facility during the last 6 y. We found that B6C3F1 mice with swollen ampullae show almost 3-fold higher pregnancy rates than mice with nonswollen ampullae when either freshly isolated or frozen-thawed embryos are implanted. Implantation of freshly collected embryos in recipients with swollen ampullae led to significantly higher pregnancy rates in comparison to implantation of frozen-thawed embryos, regardless of whether the latter were fertilized in vivo or in vitro. Moreover, we found a significant effect of genetic background on the birth rate; C57BL/6J mice and mice with a mixed genetic background had 34% higher birth rates than did C57BL/6N mice. Within the C57BL/6J group, the birth rates were significantly higher when using fresh in vivo-fertilized embryos, and cryopreservation negatively affected both in vivo- and in vitro-fertilized embryos. The success rate of obtaining one living pup was not significantly different between frozen-thawed and fresh embryos. Overall, a swollen ampulla is a strong indicator for a successful pregnancy, together with the embryo manipulation and genetic background. A better understanding of the factors that affect the reproductive outcome might lead to optimization of the ART protocols and contribute to a reduction in the number of mice used for these procedures.

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American Society of Laboratory Animal Practitioners Position Statement: Handling and Physical Restraint of Research Animals. American Society of Laboratory Animal Practitioners Position Statement: Definition of Animal Welfare. Effect of Novel High-fat Diet Feeding Methods on Food Wastage, Weight Gain, Hair Coat Grease Accumulation, and Scratching Behavior in C57BL/6NCrl Mice. Identification and Treatment of Fur Mites (Radfordia lemnina) in California Deer Mice (Peromyscus californicus) Using Selamectin. American Society of Laboratory Animal Practitioners Position Statement: Animal Care Principles.
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