Bermejo-Alvarez Pablo, Park Ki-Eun, Telugu Bhanu P
Animal Bioscience and Biotechnology Laboratory, United States Department of Agriculture; Department of Animal and Avian Sciences, University of Maryland;
Animal Bioscience and Biotechnology Laboratory, United States Department of Agriculture; Department of Animal and Avian Sciences, University of Maryland.
J Vis Exp. 2014 Feb 28(84):e51214. doi: 10.3791/51214.
The transfer of preimplantation embryos to a surrogate female is a required step for the production of genetically modified mice or to study the effects of epigenetic alterations originated during preimplantation development on subsequent fetal development and adult health. The use of an effective and consistent embryo transfer technique is crucial to enhance the generation of genetically modified animals and to determine the effect of different treatments on implantation rates and survival to term. Embryos at the blastocyst stage are usually transferred by uterine transfer, performing a puncture in the uterine wall to introduce the embryo manipulation pipette. The orifice performed in the uterus does not close after the pipette has been withdrawn, and the embryos can outflow to the abdominal cavity due to the positive pressure of the uterus. The puncture can also produce a hemorrhage that impairs implantation, blocks the transfer pipette and may affect embryo development, especially when embryos without zona are transferred. Consequently, this technique often results in very variable and overall low embryo survival rates. Avoiding these negative effects, utero-tubal embryo transfer take advantage of the utero-tubal junction as a natural barrier that impedes embryo outflow and avoid the puncture of the uterine wall. Vasectomized males are required for obtaining pseudopregnant recipients. A technique to perform vasectomy is described as a complement to the utero-tubal embryo transfer.
将植入前胚胎移植到代孕母鼠体内是生产转基因小鼠或研究植入前发育过程中发生的表观遗传改变对后续胎儿发育和成年健康影响的必要步骤。使用有效且一致的胚胎移植技术对于提高转基因动物的产生率以及确定不同处理对着床率和足月存活率的影响至关重要。囊胚期胚胎通常通过子宫移植进行移植，即在子宫壁上穿刺以插入胚胎操作吸管。吸管抽出后，子宫上形成的孔不会闭合，由于子宫的正压，胚胎可能会流入腹腔。穿刺还可能导致出血，这会损害着床、堵塞移植吸管并可能影响胚胎发育，尤其是在移植无透明带胚胎时。因此，这种技术常常导致胚胎存活率非常不稳定且总体较低。为避免这些负面影响，子宫输卵管胚胎移植利用子宫输卵管交界处作为天然屏障来阻止胚胎流出，并避免子宫壁穿刺。需要对雄性进行输精管切除术以获得假孕受体。文中描述了一种进行输精管切除术的技术，作为子宫输卵管胚胎移植的补充。
