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Dextran 玻璃化液可防止兔胚胎的黏液层和透明带损伤。

Dextran vitrification media prevents mucin coat and zona pellucida damage in rabbit embryo.

机构信息

Centro de Investigación y Tecnología Animal (CITA), Instituto Valenciano de Investigaciones Agrarias (IVIA), 12.400 Segorbe (Castellón), Spain.

出版信息

Theriogenology. 2010 Dec;74(9):1623-8. doi: 10.1016/j.theriogenology.2010.06.034. Epub 2010 Aug 12.

Abstract

Vitrification of embryos is being increasingly important for cryopreservation in mammals. However, damage and toxicity has to be reduced even more. The composition of cryoprotective medium used to immerse the embryos affects viability and developmental potential. The aim of this work was to assess the effect of the Polyvinylalcohol-PVA- and Dextran addition to vitrification media on the in vitro development of rabbit embryos from superovulated and non-superovulated females. Superovulation group were treated intramuscularly with 25 IU rhFSH. The vitrification media contained the same permeable cryoprotectans (Ethylene Glycol-ET- and Dimethyl Sulfoxide-Me₂SO-) and different macromolecules (PVA and Dextran) in different combinations. There was a significantly higher proportion of embryos without damages in mucin coat or zona pellucida after warming (undamaged embryos) in the control than in the superovulation group (95.8% vs. 83.2%, respectively). The proportion of undamaged embryos was significantly affected by the vitrification solution composition. The rate of undamaged embryos after warming in media containing 20% Me₂SO was significantly lower in media supplemented with PVA than in media with dextran (67.3 vs. 93.8, respectively). However, the proportion of undamaged embryos for the medium supplemented with dextran was similar for media with 15 or 20% Me₂SO. In conclusion, the addition of dextran to the vitrification media improve the preservation of rabbit embryos and permits to reduce the amount of Me₂SO for vitrification. Additionally, in vitro developmental ability of undamaged embryos were not affected by superovulation treatment nor vitrification media.

摘要

胚胎玻璃化冷冻保存在哺乳动物中越来越重要。然而,还需要进一步减少损伤和毒性。用于浸泡胚胎的冷冻保护剂的组成会影响胚胎的活力和发育潜能。本研究旨在评估向玻璃化液中添加聚乙烯醇(PVA)和葡聚糖对超排卵和未超排卵母兔胚胎体外发育的影响。超排卵组肌肉内注射 25IU rhFSH。玻璃化液中含有相同的可渗透冷冻保护剂(乙二醇(EG)和二甲基亚砜(DMSO))和不同的大分子(PVA 和葡聚糖),以不同的组合存在。在解冻后(无损伤胚胎),黏液层或透明带无损伤的胚胎比例在对照组中明显高于超排卵组(分别为 95.8%和 83.2%)。无损伤胚胎的比例明显受到玻璃化溶液组成的影响。在含有 20% DMSO 的玻璃化液中,添加 PVA 的玻璃化液中无损伤胚胎的比例明显低于添加葡聚糖的玻璃化液(分别为 67.3%和 93.8%)。然而,对于添加葡聚糖的玻璃化液,添加 15%或 20% DMSO 的玻璃化液中无损伤胚胎的比例相似。总之,向玻璃化液中添加葡聚糖可提高兔胚胎的保存效果,并允许减少玻璃化所需的 DMSO 量。此外,超排卵处理和玻璃化液均不影响无损伤胚胎的体外发育能力。

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