Gasparrini Bianca, Attanasio Laura, De Rosa Anna, Monaco Elisa, Di Palo Rossella, Campanile Giuseppe
DISCIZIA, Faculty of Veterinary Medicine, Federico II University, Via Delpino 1, 80137 Naples, Italy.
Anim Reprod Sci. 2007 Apr;98(3-4):335-42. doi: 10.1016/j.anireprosci.2006.04.046. Epub 2006 Apr 26.
The aim of this study was to evaluate the efficiency of the solid surface vitrification (SSV) and the cryoloop vitrification (CLV) methods to cryopreserve in vitro matured buffalo oocytes. Another objective of the work was to investigate whether the presence of cumulus cells affects the efficiency of oocyte vitrification in this species. In the SSV method, oocytes were vitrified in a solution of 35% ethylene glycol, 5% polyvinyl-pyrrolidone and 0.4% trehalose and they were warmed in a 0.3M trehalose solution. In the CLV method, oocytes were vitrified in 16.5% ethylene glycol and 16.5% dimethyl sulfoxide and warmed in decreasing concentrations of sucrose. The oocytes that survived vitrification were fertilized and cultured in vitro up to the blastocyst stage. Although high survival rates were recorded in all groups, when the oocytes were vitrified by the CLV method in the absence of cumulus cells, the survival rate was significantly (P<0.05) lower. However, the CLV gave a significantly higher cleavage rate compared to the SSV with the denuded oocytes (45% versus 26%, respectively; P<0.05), whereas no differences were found between methods with the cumulus-enclosed oocytes (14% versus 15%, respectively). Blastocysts were produced for the first time from in vitro matured oocytes that were vitrified-warmed in buffalo. Nevertheless, vitrification significantly decreased blastocyst yield, regardless of both the method employed and the presence or absence of cumulus cells.
本研究的目的是评估固体表面玻璃化法（SSV）和冷冻环玻璃化法（CLV）冷冻体外成熟水牛卵母细胞的效率。该研究的另一个目的是调查卵丘细胞的存在是否会影响该物种卵母细胞玻璃化的效率。在SSV方法中，卵母细胞在含有35%乙二醇、5%聚乙烯吡咯烷酮和0.4%海藻糖的溶液中进行玻璃化处理，然后在0.3M海藻糖溶液中复温。在CLV方法中，卵母细胞在16.5%乙二醇和16.5%二甲基亚砜中进行玻璃化处理，并在浓度递减的蔗糖溶液中复温。将玻璃化后存活的卵母细胞进行体外受精并培养至囊胚阶段。尽管所有组的存活率都很高，但当卵母细胞在无卵丘细胞的情况下采用CLV方法进行玻璃化处理时，存活率显著降低（P<0.05）。然而，对于裸卵，CLV的分裂率显著高于SSV（分别为45%和26%；P<0.05），而对于有卵丘包裹的卵母细胞，两种方法之间没有差异（分别为14%和15%）。首次从经玻璃化-复温处理的体外成熟水牛卵母细胞中获得了囊胚。然而，无论采用何种方法以及有无卵丘细胞，玻璃化都会显著降低囊胚产量。
