Morató Roser, Izquierdo Dolors, Albarracín José Luis, Anguita Begoña, Palomo María Jesús, Jiménez-Macedo Ana Raquel, Paramio María Teresa, Mogas Teresa
Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Mol Reprod Dev. 2008 Jan;75(1):191-201. doi: 10.1002/mrd.20725.
The purpose of this study was to determine the efficacy of pre-treating mature bovine oocytes with Taxol before vitrification by the open pulled Straw method (OPS). We evaluated the effects of pre-treating the oocytes with 1 microM Taxol on chromosome organization, spindle morphology, cortical granule distribution and the ability of fertilized oocytes to develop to the blastocyst stage. After calf or cow oocyte vitrification without Taxol, significantly higher proportions of spindle abnormalities in the form of abnormal spindle structures or dispersed or decondensed chromosomes were observed compared to fresh control oocytes. In contrast, when we compared calf oocytes pre-treated with Taxol before vitrification with control calf oocytes, similar percentages of oocytes showing a normal spindle morphology were observed. The percentages of oocytes with a peripheral cortical granule (CG) distribution increased when the oocytes were pretreated with Taxol and vitrified, while oocytes vitrified without Taxol pre-treatment gave rise to higher cortical distribution percentages. Cleavage and blastocyst rates were significantly lower for vitrified versus untreated oocytes, both in cow and calf oocytes. Significantly higher cleavage rates were obtained when calf and cow oocytes were vitrified with Taxol. Pre-treatment with Taxol before cow oocyte vitrification yielded significantly higher blastocyst rates. Calf oocytes, however, were unable to develop to the blastocyst stage, irrespective of previous Taxol treatment. These results indicate that the pre-treatment of oocytes with Taxol before vitrification helps to reduce the damage induced by the cryopreservation process, and potentially improves the subsequent development of vitrified bovine oocytes. Summary sentence: Pre-treatment of oocytes with Taxol before vitrification helps to reduce the damage induced by vitrification and potentially improves the development of vitrified bovine oocytes.
本研究的目的是确定通过开放式拉长细管法（OPS）在玻璃化之前用紫杉醇预处理成熟牛卵母细胞的效果。我们评估了用1微摩尔紫杉醇预处理卵母细胞对染色体组织、纺锤体形态、皮质颗粒分布以及受精卵发育至囊胚阶段能力的影响。在未用紫杉醇进行小牛或母牛卵母细胞玻璃化后，与新鲜对照卵母细胞相比，观察到以异常纺锤体结构、分散或解聚染色体形式存在的纺锤体异常比例显著更高。相比之下，当我们将玻璃化前用紫杉醇预处理的小牛卵母细胞与对照小牛卵母细胞进行比较时，观察到显示正常纺锤体形态的卵母细胞百分比相似。在用紫杉醇预处理并玻璃化的卵母细胞中，具有外周皮质颗粒（CG）分布的卵母细胞百分比增加，而未进行紫杉醇预处理而玻璃化的卵母细胞则产生更高的皮质分布百分比。在母牛和小牛卵母细胞中，玻璃化的卵母细胞的分裂率和囊胚率均显著低于未处理的卵母细胞。当小牛和母牛卵母细胞用紫杉醇进行玻璃化时，获得了显著更高的分裂率。在母牛卵母细胞玻璃化前用紫杉醇预处理产生了显著更高的囊胚率。然而，无论之前是否进行紫杉醇处理，小牛卵母细胞都无法发育至囊胚阶段。这些结果表明，在玻璃化之前用紫杉醇预处理卵母细胞有助于减少冷冻保存过程引起的损伤，并可能改善玻璃化牛卵母细胞的后续发育。总结句：在玻璃化之前用紫杉醇预处理卵母细胞有助于减少玻璃化引起的损伤，并可能改善玻璃化牛卵母细胞的发育。
