Martinez Emilio A, Angel Miguel Angel, Cuello Cristina, Sanchez-Osorio Jonatan, Gomis Jesus, Parrilla Inmaculada, Vila Jordi, Colina Ignaci, Diaz Marta, Reixach Josep, Vazquez Jose Luis, Vazquez Juan Maria, Roca Jordi, Gil Maria Antonia
Department of Animal Medicine and Surgery, University of Murcia, Murcia, Spain.
Department of Research and Development, Selección Batallé S.A., Girona, Spain.
PLoS One. 2014 Aug 13;9(8):e104696. doi: 10.1371/journal.pone.0104696. eCollection 2014.
Excellent fertility and prolificacy have been reported after non-surgical deep uterine transfers of fresh in vivo-derived porcine embryos. Unfortunately, when this technology is used with vitrified embryos, the reproductive performance of recipients is low. For this reason and because the embryos must be stored until they are transferred to the recipient farms, we evaluated the potential application of non-surgical deep uterine transfers with in vivo-derived morulae cultured for 24 h in liquid stage. In Experiment 1, two temperatures (25 °C and 37 °C) and two media (one fully defined and one semi-defined) were assessed. Morulae cultured in culture medium supplemented with bovine serum albumin and fetal calf serum at 38.5 °C in 5% CO2 in air were used as controls. Irrespective of medium, the embryo viability after 24 h of culture was negatively affected (P<0.05) at 25 °C but not at 37 °C compared with the controls. Embryo development was delayed in all experimental groups compared with the control group (P<0.001). Most of the embryos (95.7%) cultured at 37 °C achieved the full or expanded blastocyst stage, and unlike the controls, none of them hatched at the end of culture. In Experiment 2, 785 morulae were cultured in the defined medium at 37 °C for 24 h, and the resulting blastocysts were transferred to the recipients (n = 24). Uncultured embryos collected at the blastocyst stage (n = 750) were directly transferred to the recipients and used as controls (n = 25). No differences in farrowing rates (91.7% and 92.0%) or litter sizes (9.0 ± 0.6 and 9.4 ± 0.8) were observed between the groups. This study demonstrated, for the first time, that high reproductive performance can be achieved after non-surgical deep uterine transfers with short-term cultured morulae in a defined medium, which opens new possibilities for the sanitary, safe national and international trade of porcine embryos and the commercial use of embryo transfer in pigs.
据报道,对体内新鲜获取的猪胚胎进行非手术子宫深部移植后,生育力和繁殖力良好。不幸的是,当该技术用于玻璃化胚胎时,受体的繁殖性能较低。出于这个原因,也因为胚胎必须储存至转移到受体农场,我们评估了对在液体阶段培养24小时的体内桑葚胚进行非手术子宫深部移植的潜在应用。在实验1中,评估了两种温度(25℃和37℃)和两种培养基(一种完全限定培养基和一种半限定培养基)。在添加牛血清白蛋白和胎牛血清的培养基中于38.5℃、5%二氧化碳的空气中培养的桑葚胚用作对照。与对照相比,无论培养基如何,在25℃下培养24小时后胚胎活力均受到负面影响(P<0.05),而在37℃下则未受影响。与对照组相比,所有实验组的胚胎发育均延迟(P<0.001)。在37℃下培养的大多数胚胎(95.7%)达到了完全或扩张囊胚阶段,与对照组不同的是,培养结束时它们均未孵化。在实验2中,785个桑葚胚在限定培养基中于37℃培养24小时,将得到的囊胚转移至受体(n = 24)。在囊胚阶段收集的未培养胚胎(n = 750)直接转移至受体并用作对照(n = 25)。两组之间的产仔率(91.7%和92.0%)或窝产仔数(9.0±0.6和9.4±0.8)未观察到差异。本研究首次证明,对在限定培养基中短期培养的桑葚胚进行非手术子宫深部移植后可实现高繁殖性能,这为猪胚胎的卫生、安全的国内和国际贸易以及猪胚胎移植的商业应用开辟了新的可能性。