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在冷却或玻璃化之前暴露于甲基-β-环糊精对牛未成熟卵母细胞活力的影响。

Effect of the exposure to methyl-β-cyclodextrin prior to chilling or vitrification on the viability of bovine immature oocytes.

机构信息

University of Brasília, Brasília, DF, Brazil.

出版信息

Cryobiology. 2012 Dec;65(3):319-25. doi: 10.1016/j.cryobiol.2012.09.001. Epub 2012 Sep 13.

Abstract

The present study aimed to evaluate the effect of methyl-β-cyclodextrin (MβCD) as a cholesterol loader to change oocyte plasma membrane and increase its tolerance toward cryopreservation. The first and second experiments were conducted to investigate if MβCD could improve nuclear and cytoplasmic maturation after oocyte exposure to cold stress for 10 or 30 min, respectively. No differences (P>0.05) in either experiment in the metaphase II (MII) rate of oocytes exposed to MβCD and cold stress; but these oocytes presented lower maturation rates than control groups. In the second experiment, a lower percentage of oocytes showed degenerated chromatin (P<0.05) after exposure to 2mg/mL of MβCD compared to the group exposed to 0mg/mL. However, no differences among treatments were observed in cytoplasmic maturation. Groups exposed to cold stress demonstrated a lower (P<0.05) capacity for embryonic development compared to the control groups. In the third experiment immature oocytes were exposed to MβCD and then, vitrified (cryotop). After warming, we observed that the ability to reach MII and chromatin degeneration were altered (P<0.05) by MβCD. The blastocysts rate (P<0.05) on D7 was higher in the 2 mg/mL MβCD group, but an identical finding was not observed on D8 (P>0.05). Chromatin degeneration was higher in the vitrification groups. We conclude that MβCD improved nuclear maturation by reducing oocyte degeneration after cold stress or vitrification; however, more studies are required to clarify the usefulness of MβCD use in oocyte cryopreservation.

摘要

本研究旨在评估甲基-β-环糊精(MβCD)作为胆固醇载体对改变卵母细胞质膜并提高其对冷冻保存的耐受性的影响。第一和第二实验分别研究了 MβCD 是否可以改善卵母细胞暴露于冷应激 10 或 30 分钟后的核和细胞质成熟。在这两个实验中,暴露于 MβCD 和冷应激的卵母细胞的中期 II(MII)率没有差异(P>0.05);但这些卵母细胞的成熟率低于对照组。在第二个实验中,与暴露于 0mg/mL 的组相比,暴露于 2mg/mL 的 MβCD 的卵母细胞显示出较低百分比的退化染色质(P<0.05)。然而,在细胞质成熟方面,各组之间没有差异。暴露于冷应激的组的胚胎发育能力低于对照组(P<0.05)。在第三个实验中,不成熟的卵母细胞暴露于 MβCD ,然后进行玻璃化(cryotop)。在解冻后,我们观察到 MβCD 改变了达到 MII 和染色质退化的能力(P<0.05)。在第 7 天,2mg/mL MβCD 组的囊胚率(P<0.05)较高,但在第 8 天没有观察到相同的结果(P>0.05)。玻璃化组的染色质退化更高。我们得出结论,MβCD 通过减少冷应激或玻璃化后卵母细胞的退化来改善核成熟;然而,需要进一步的研究来阐明 MβCD 在卵母细胞冷冻保存中的有用性。

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