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[人卵母细胞玻璃化冷冻保存时间及解冻方法对辅助生殖结局的影响]

[Effects of cryopreservation time and thawing method of human oocyte vitrification on the outcome of assisted reproduction].

作者信息

Song Wen-yan, Sun Ying-pu, Jin Hai-xia, Xin Zhi-min, Su Ying-chun, Guo Yi-hong, Chen Zi-jiang

机构信息

Reproductive Medical Center, First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.

出版信息

Zhonghua Fu Chan Ke Za Zhi. 2010 Aug;45(8):578-82.

Abstract

OBJECTIVE

To evaluate the effects on pregnancy outcome of freezing time from oocyte retrieval and thawing method for metaphaseII human oocytes vitrification.

METHODS

From Mar 2007 to Mar 2009, the clinical outcome of 30 infertile women undergoing vitrified-thawing oocytes of in vitro fertilization-embryo transfer (IVF-ET) in the Reproductive Medical Center of the First Affiliated Hospital of Zhengzhou University was studied retrospectively, including 21 women with double fallopian tube obstruction and 9 women's husband azoospermia. All infertile women were divided into three groups, including 5 cases in group A (freezing between 4 and 5 hours from oocyte retrieval and conventional thawing method), 9 cases in group B (freezing within 2 hours from retrieval and conventional thawing method) and 16 cases in group C (freezing within 2 hours from retrieval and improved thawing method). The vitrified oocytes were preserved for 2 months to 1 year and thawed for Intracytoplasmic sperm injection (ICSI) and embryo transfer. The outcome of IVF and pregnancy were recorded.

RESULTS

(1) The rates of oocyte survival was (65 ± 33)% in group B and (72 ± 23)% in group C and the rate of transfer cycle was 9/9 in group B and 16/16 in group C, which were all significantly higher than (16 ± 17)% of oocyte survival and 1/5 of transfer cycle in group A (P = 0.001, 0.021). However, the rate of oocyte survival and transfer cycle between group B and group C did not reach statistical difference (P > 0.05). The rate of implantation and clinical pregnancy of (33 ± 38)% and 9/16 in group C were significantly higher (4 ± 11)% and 1/9 in group B (P = 0.033, 0.040). (2) The mean age of women in group C were (28.6 ± 2.1) in oneself oocyte, (28.0 ± 4.6) in donor oocyte and (28.1 ± 3.4) in donor sperm. The rate of oocyte survival was (73 ± 25)%, (88 ± 10)% and (66 ± 25)%. The rate of fertilization rate was (84.6 ± 0.9)%, (79.3 ± 2.0)% and (82.8 ± 15.0)%. The rate of implantation was (20.0 ± 44.7)%, (33.0 ± 0.1)%, (41.6 ± 41.7)%. The rate of clinical pregnancy was 1/5 in oneself cycles, 3/3 in donor oocyte cycles, 5/8 banked donor sperm cycles in group C. All above clinical parameters were not statistically different (P > 0.05). (3) In group A, one women underwent IVF-ET and no clinical pregnancy was observed. One women pregnancy was terminated at two months in group B. The clinical pregnancies rate of group C was 9/16, late abortion occurred in 1 woman, the other 8 women underwent term pregnancy, including 5 male infants and 4 female infants. All of infants showed normal Karyotype. Live-birth rates per warmed oocyte was 5.9%(8/135). The mean gestational weeks and birth weight of the infants were (39.4 ± 0.9) weeks and (3574 ± 569) g, respectively.

CONCLUSIONS

Embryo quality and clinical outcome of thawing cycles could be significantly improved when oocyte vitrification was performed within 2 hours from oocyte retrieval and improved thawing method.

摘要

目的

评估中期Ⅱ人卵母细胞玻璃化冷冻的取卵后冷冻时间及解冻方法对妊娠结局的影响。

方法

回顾性分析2007年3月至2009年3月在郑州大学第一附属医院生殖医学中心接受体外受精-胚胎移植(IVF-ET)玻璃化冻融卵母细胞的30例不孕妇女的临床结局,其中双侧输卵管阻塞21例,男方无精子症9例。所有不孕妇女分为三组,A组5例(取卵后4至5小时冷冻,采用传统解冻方法),B组9例(取卵后2小时内冷冻,采用传统解冻方法),C组16例(取卵后2小时内冷冻,采用改良解冻方法)。玻璃化卵母细胞保存2个月至1年,解冻后行卵胞浆内单精子注射(ICSI)及胚胎移植。记录IVF及妊娠结局。

结果

(1)B组卵母细胞存活率为(65±33)%,C组为(72±23)%,B组移植周期率为9/9,C组为16/16,均显著高于A组的卵母细胞存活率(16±17)%及移植周期率1/5(P=0.001,0.021)。然而,B组与C组之间的卵母细胞存活率及移植周期率未达到统计学差异(P>0.05)。C组的着床率及临床妊娠率分别为(33±38)%及9/16,显著高于B组的(4±11)%及1/9(P=0.033,0.040)。(2)C组自身卵母细胞的妇女平均年龄为(28.6±2.1)岁,供体卵母细胞为(28.0±4.6)岁,供体精子为(28.1±3.4)岁。卵母细胞存活率分别为(73±25)%、(88±10)%及(66±25)%。受精率分别为(84.6±0.9)%、(79.3±2.0)%及(82.8±15.0)%。着床率分别为(20.0±44.7)%、(33.0±0.1)%、(41.6±41.7)%。C组自身周期临床妊娠率为1/5,供体卵母细胞周期为3/3,供体精子库周期为5/8。上述所有临床参数均无统计学差异(P>0.05)。(3)A组1例妇女接受IVF-ET,未观察到临床妊娠。B组1例妇女妊娠2个月时终止妊娠。C组临床妊娠率为9/16,1例妇女发生晚期流产,其他8例妇女足月妊娠,其中男婴5例,女婴4例。所有婴儿核型均正常。每个解冻卵母细胞的活产率为5.9%(8/135)。婴儿的平均孕周及出生体重分别为(39.4±0.9)周及(3574±569)g。

结论

取卵后2小时内进行卵母细胞玻璃化冷冻并采用改良解冻方法可显著改善解冻周期的胚胎质量及临床结局。

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