Pereira R M, Marques C C
Departamento de Reprodução Animal, Estação Zootécnica Nacional (EZN), 2005-048, Vale de Santarem, Portugal.
Cell Tissue Bank. 2008 Dec;9(4):267-77. doi: 10.1007/s10561-008-9075-2. Epub 2008 May 22.
Cryopreservation of oocytes and embryos is a crucial step for the widespread and conservation of animal genetic resources. However, oocytes and early embryos are very sensitive to chilling and cryopreservation and although new advances have been achieved in the past few years the perfect protocol has not yet been established. All oocytes and embryos suffer considerable morphological and functional damage during cryopreservation but the extent of the injury as well as differences in survival and developmental rates may be highly variable depending on the species, developmental stage and origin (for example, in vitro produced or in vivo derived, micromanipulated or not). Currently, there are two methods for gamete and embryos cryopreservation: slow freezing and vitrification. We have experienced both techniques but vitrification has become a viable and promising alternative to traditional approaches especially when dealing with in vitro produced or micromanipulated embryos and oocytes. Recently new strategies based on emerging studies in the field of lipid research have been used to reduce intracellular lipid content in bovine in vitro produced embryos and therefore increase their tolerance to micromanipulation and cryopreservation. The addition of a conjugated isomer of linoleic acid, the trans-10, cis-12 octadecadienoic acid to embryo culture medium more than twice improved embryo post-thawing viability after micromanipulation and vitrification. Vitrification was also used for the cryopreservation of embryos belonging to the Portuguese Animal Germplasm Bank project presently running at our facilities.
卵母细胞和胚胎的冷冻保存是动物遗传资源广泛保存的关键步骤。然而,卵母细胞和早期胚胎对低温和冷冻保存非常敏感,尽管在过去几年中取得了新进展,但尚未建立完善的方案。所有卵母细胞和胚胎在冷冻保存过程中都会遭受相当程度的形态和功能损伤,但损伤程度以及存活和发育率的差异可能因物种、发育阶段和来源(例如,体外产生或体内衍生、是否经过显微操作)而有很大差异。目前,有两种用于配子和胚胎冷冻保存的方法:慢速冷冻和玻璃化。我们对这两种技术都有经验,但玻璃化已成为传统方法的一种可行且有前景的替代方法,特别是在处理体外产生或经过显微操作的胚胎和卵母细胞时。最近,基于脂质研究领域新出现的研究的新策略已被用于降低牛体外产生胚胎的细胞内脂质含量,从而提高它们对显微操作和冷冻保存的耐受性。在胚胎培养基中添加亚油酸的共轭异构体反式-10,顺式-12十八碳二烯酸,可使显微操作和玻璃化后的胚胎解冻后活力提高两倍以上。玻璃化还用于目前在我们设施中运行的葡萄牙动物种质库项目的胚胎冷冻保存。