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使用Cryotop玻璃化冷冻法与小鼠卵母细胞的成熟、受精及发育率

Vitrification by Cryotop and the Maturation, Fertilization, and Developmental Rates of Mouse Oocytes.

作者信息

Abedpour Neda, Rajaei Farzad

机构信息

Cellular and Molecular Research Centre, Qazvin University of Medical Sciences, Qazvin, IR Iran.

Cellular and Molecular Research Centre, Qazvin University of Medical Sciences, Qazvin, IR Iran ; Fertility and Infertility Research Centre, Kosar Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran.

出版信息

Iran Red Crescent Med J. 2015 Oct 6;17(10):e18172. doi: 10.5812/ircmj.18172. eCollection 2015 Oct.

Abstract

BACKGROUND

Oocyte cryopreservation is an important part of modern fertility treatment. The effect of vitrification on the fertilization and developmental rates of embryo is still a matter of debate.

OBJECTIVES

This study aimed to investigate the effect of vitrification on the success of mouse oocyte maturation, fertilization, and preimplantation development in vitro.

MATERIALS AND METHODS

In this experimental study, a total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into two control and experimental (vitrified) groups. Oocytes in the experimental group were vitrified by Cryotop using vitrification medium (Origio, Denmark) and kept in liquid nitrogen for one month. Then, they were cultured in maturation medium for 24 hours. In vitro maturated metaphase 2 (IVM-MII) and ovulated metaphase 2 (OV-MII) oocytes were inseminated and the fertilized embryos assessed until the hatching blastocyst stage. Outcomes were assessed for statistical significance by Chi-square test using SPSS software.

RESULTS

Vitrification caused a significant reduction in the maturation rate of oocytes. Of those that matured, the fertilization rate of vitrified IVM-MII (44.1%) and OV-MII oocytes (50%) was not significantly different from each other but both were significantly lower than the control group (P < 0.05). There was no significant difference in developmental rates of both vitrified groups and the control group.

CONCLUSIONS

The present study showed that vitrification using Cryotop and freezing medium can damage oocytes by reducing the maturation and fertilization rates in both developmental stages.

摘要

背景

卵母细胞冷冻保存是现代生育治疗的重要组成部分。玻璃化对胚胎受精率和发育率的影响仍存在争议。

目的

本研究旨在探讨玻璃化对小鼠卵母细胞体外成熟、受精和植入前发育成功率的影响。

材料与方法

在本实验研究中,分别从NMRI小鼠的卵巢和输卵管中获取了200个生发泡(GV)卵母细胞和200个中期II(MII)卵母细胞,并分为两个对照组和实验组(玻璃化组)。实验组的卵母细胞使用玻璃化培养基(丹麦Origio公司)通过Cryotop进行玻璃化处理,并在液氮中保存1个月。然后,将它们在成熟培养基中培养24小时。体外成熟的中期2(IVM-MII)和排卵的中期2(OV-MII)卵母细胞进行受精,并对受精后的胚胎进行评估,直至囊胚孵化阶段。使用SPSS软件通过卡方检验评估结果的统计学意义。

结果

玻璃化导致卵母细胞成熟率显著降低。在成熟的卵母细胞中,玻璃化的IVM-MII(44.1%)和OV-MII卵母细胞(50%)的受精率彼此之间无显著差异,但均显著低于对照组(P < 0.05)。玻璃化组和对照组的发育率均无显著差异。

结论

本研究表明,使用Cryotop和冷冻培养基进行玻璃化处理会降低两个发育阶段的成熟率和受精率,从而损害卵母细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de7d/4636753/e1d7b1f8b57a/ircmj-17-10-18172-g001.jpg

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