Cuello Cristina, Martinez Cristina A, Nohalez Alicia, Parrilla Inmaculada, Roca Jordi, Gil Maria A, Martinez Emilio A
Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Murcia, E-3100, Murcia, Spain.
Sci Rep. 2016 Sep 26;6:33915. doi: 10.1038/srep33915.
The use of pH-stable media would simplify embryo vitrification and the warming of porcine embryos and might facilitate the application of embryo transfer in practice. In this work, we investigated whether a pH-stable basal medium constituted of Tyrode's lactate medium, polyvinyl alcohol, and HEPES for buffering was suitable for porcine embryo vitrification warming in place of the conventional gas-equilibrated media. A high percentage (>90%) of embryos survived vitrification and warming in this medium, achieving in vitro survival rates similar to embryos vitrified-warmed using the conventional protocol and their fresh counterparts. The pH-stable medium did not affect the in vivo developmental competence of the vitrified-warmed embryos. A farrowing rate of 71.4% (5/7) with 10.4 ± 3.1 piglets born was obtained for the embryos vitrified and warmed in this medium and transferred to selected recipients. This medium will enable the use of simple, safe and standardized protocols for the vitrification and warming of porcine embryos for optimal embryo survival and quality when applied under field conditions. This study opens new possibilities for the widespread use of embryo transfer in pigs.
使用pH稳定的培养基将简化猪胚胎的玻璃化冷冻及解冻过程,并可能促进胚胎移植在实际中的应用。在本研究中,我们探究了一种由乳酸盐Tyrode培养基、聚乙烯醇和用于缓冲的HEPES组成的pH稳定基础培养基,是否适合替代传统的气体平衡培养基用于猪胚胎的玻璃化冷冻解冻。在此培养基中,超过90%的胚胎在玻璃化冷冻和解冻后存活,其体外存活率与使用传统方法玻璃化冷冻解冻的胚胎及其新鲜胚胎相当。pH稳定培养基并未影响玻璃化冷冻解冻胚胎的体内发育能力。在此培养基中进行玻璃化冷冻解冻并移植到选定受体的胚胎,产仔率为71.4%(5/7),平均每窝产仔10.4±3.1头。当在野外条件下应用时,这种培养基将能够使用简单、安全和标准化的方案进行猪胚胎的玻璃化冷冻和解冻,以实现最佳的胚胎存活和质量。本研究为猪胚胎移植的广泛应用开辟了新的可能性。