Assisted Reproduction Technologies and Cryopreservation Unit, CMB/National Institute of Allergy and Infectious Diseases/ National Institutes of Health, Rockville, Maryland, USA.
Theriogenology. 2011 Sep 1;76(4):729-36. doi: 10.1016/j.theriogenology.2011.04.005. Epub 2011 May 23.
In vitro fertilization (IVF) is one of the most important techniques used for assisted reproduction in mouse colony management. As with natural mating, where mice have varying fertility indices, fertility rates of genetically modified (GM) [transgenic (Tg), knock out (KO) and congenic (Cg)] mice are influenced by their genetic background. Lines of GM mice that have poor fertility have a concomitant poor IVF outcome. Treatment of mouse sperm with extracellular adenosine 5'-triphosphate (ATPe) enhanced in vitro fertilization rates in outbred and hybrid mice. The objective of this study was to analyze the effects of using extracellular adenosine 5'-triphosphate-treated sperm for IVF of inbred wild type, and genetically modified mouse lines, for which standard IVF did not work well. The IVF was performed using the GM mice on C57BL/10SnJ, C57BL/6J, BALB/cJ and NFS/N background strains and wild type (WT) mice such as C57BL/6N, BALB/cAnN, and B6129SF1 strains. Oocytes from superovulated females were fertilized in vitro with sperm from the same background strain, and either treated or not treated with ATPe. The ATPe treatment enhanced IVF outcome in most of the GM and some WT strains, as indicated by the percentage of embryos that progressed to the two-cell stage. There was no marked difference between ATPe treated and control groups for the development rate of two-cell embryos to blastocysts in culture, or in the number of pups born after transfer of two-cell embryos into recipient females. The observed improvement of the IVF results following ATPe treatment of transgenic and KO mouse sperm were a potential solution for improving the outcome of assisted reproduction techniques used for rederivation or for gamete banking.
体外受精（IVF）是小鼠群体管理中辅助生殖的最重要技术之一。与自然交配一样，由于小鼠的生育指数各不相同，因此基因修饰（GM）[转基因（Tg）、敲除（KO）和同系（Cg）]小鼠的生育能力受到其遗传背景的影响。生育能力差的 GM 小鼠系同时也具有较差的 IVF 结果。用细胞外腺苷 5'-三磷酸（ATPe）处理小鼠精子可提高杂交和杂种小鼠的体外受精率。本研究的目的是分析使用细胞外腺苷 5'-三磷酸处理的精子进行 IVF 的效果，这些精子用于标准 IVF 效果不佳的近交野生型和基因修饰的小鼠系。使用 GM 小鼠在 C57BL/10SnJ、C57BL/6J、BALB/cJ 和 NFS/N 背景品系以及 C57BL/6N、BALB/cAnN 和 B6129SF1 等野生型（WT）小鼠上进行 IVF。用来自同一背景品系的精子对超排卵雌性的卵母细胞进行体外受精，并对精子进行或不进行 ATPe 处理。在大多数 GM 和一些 WT 品系中，ATPe 处理增强了 IVF 结果，表现为胚胎发育到二细胞阶段的百分比增加。在培养中二细胞胚胎的发育率或将二细胞胚胎转移到受体雌性后出生的幼仔数量方面，ATPe 处理组和对照组之间没有明显差异。在转基和 KO 小鼠精子中，用 ATPe 处理提高 IVF 结果的效果为改进用于再衍生或配子库存储的辅助生殖技术的结果提供了潜在的解决方案。
