冷冻载体及蔗糖浓度对复苏卵母细胞存活率的影响

doi:10.3969/j.issn.1004-616x.2012.06.012

癌变·畸变·突变

冷冻载体及蔗糖浓度对复苏卵母细胞存活率的影响

张红琴/刘思瑶/黄军/孟祥黔/艾玲/雷婉秋/钟影*

四川省成都市锦江妇幼保健院，四川成都 610016

收稿日期:2012-08-30 修回日期:2012-09-21 出版日期:2011-11-30 发布日期:2011-11-30
通讯作者: 钟影,-mail：yzhong08@126.com
作者简介:张红琴 (1980－ )，女，山西人，硕士，研究方向：胚胎工程。
基金资助:
四川省卫生厅科学研究基金项目 (090099)

The effects of the different combinations of freezing carrier and sucrose concentration on the oocyte survival after thawing

ZHANG Hong-qin，LIU Si-yao，HUANG Jun，MENG Xiang-qian，AI Ling，LEI Wan-qiu，ZHONG Ying*

Center for Reproductive Medicine, Chengdu Jinjiang Hospital for Maternal and Child Health Care, Chengdu 610016, Sichuan, China

Received:2012-08-30 Revised:2012-09-21 Online:2011-11-30 Published:2011-11-30
Contact: ZHONG Ying，E-mail：yzhong08@126.com
About author:张红琴 (1980－ )，女，山西人，硕士，研究方向：胚胎工程。
Supported by:
四川省卫生厅科学研究基金项目 (090099)

摘要/Abstract

摘要：

目的：探讨MⅡ卵母细胞玻璃化冷冻和复苏过程中不同冷冻载体和不同蔗糖浓度解冻对卵母细胞存活率的影响。方法：收集2009年6月至2012年5月在成都市锦江区妇幼保健院生殖医学中心常规IVF治疗周期第1天没有受精的MⅡ卵母细胞853枚，进行玻璃化冷冻后复苏。按照实验过程中所用不同冷冻载体和不同解冻方法分为4组。A组79枚，载体为冷冻环，采用常规方法解冻；B组580枚，载体为半麦管，采用常规方法解冻；C组121枚，载体为冷冻环，采用改良法解冻；D组73枚，载体为半麦管，采用改良法解冻。将卵母细胞复苏2 h后观察，对各组卵母细胞存活率进行了比较。结果：C组卵母细胞存活率 (97.52%)显著高于A组 (72.15%)、B组 (52.24%)和D组 (64.38%)(P< 0.01)；A组卵母细胞存活率高于B组，差异显著 (P< 0.05)；其余各组间卵母细胞存活率差异无统计学意义 (P> 0.05)。结论：采用冷冻环冷冻及改良法解冻能提高成熟卵母细胞存活率。

关键词: 蔗糖浓度, 冷冻载体, 卵母细胞, 玻璃化冷冻

Abstract:

OBJECTIVE: To investigate the effects of different combinations of sucrose concentration and freezing carrier on oocyte survival after thawing. METHODS：853 failed-fertilization oocytes after conventional IVF were collected and frozen with vitrification method in Center for Reproductive Medicine，Chengdu Jinjiang Hospital for Women and Child Health Care from June 2009 to May 2012. All oocytes were divided into four groups；group A：79 oocytes were vitrified with the cryoloop carrier and thawed using the conventional method；group B：580 oocytes were vitrified with the hemi-straw carrier and thawed using the conventional method；group C：121 oocytes were vitrified with the cryoloop carrier and thawed using the modified method；group D：73 oocytes were vitrified with the hemi-straw carrier and thawed using the modified method. After thawing for 2 h，the oocyte survival rates were compared among the groups mentioned above. RESULTS：The rate of oocyte survival in group C (97.52%) was significantly higher than those in group A (72.15%)，group B (52.24%) and group D (64.38%) (P< 0.01，respectively). The rate of oocyte survival in group A was significantly higher than that in group B (P< 0.05). No statistical difference in the rate of oocyte survival was found between other groups (P>0.05). CONCLUSION：The survival rate of MⅡ oocytes could be improved when oocytes were vitrified with the cryoloop carrier and thawed using the modified method.

Key words: sucrose concentration, frozen carrier, oocytes, vitrification

张红琴/刘思瑶/黄军/孟祥黔/艾玲/雷婉秋/钟影*. 冷冻载体及蔗糖浓度对复苏卵母细胞存活率的影响[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004-616x.2012.06.012.

ZHANG Hong-qin，LIU Si-yao，HUANG Jun，MENG Xiang-qian，AI Ling，LEI Wan-qiu，ZHONG Ying*. The effects of the different combinations of freezing carrier and sucrose concentration on the oocyte survival after thawing[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2012.06.012.

参考文献

[1] Chen C. Pregnancy after human oocyte cryopreservation [J]. Lancet， 1986，1 (8486)：884-886.

[2] Cobo A，Domingo J，Pérez S，et al. Vitrification：an effective new approach to oocyte banking and p reserving fertility in cancer patients[J]. Clin Transl Oncol，2008，10 (5)：268-273.

[3] Vajta G，Kuwayama M. Improving cryopreservation systems[J]. Theriogenology，2006，65 (1)：236-244.

[4] Mazur P. Freezing of living cells：mechanisms and implications[J]. Physiol，1984，247 (3 Sup 1)：125–142.

[5] Watson PF，Morris GJ. Cold shock injury in animal cells[J]. Symp Soc Exp Biol，1987，41：311–340.

[6] Pickering SJ，Braude PR，Johnson MH，et al. Transient cooling to room temperature can cause irreversible disruption of the meiotic spindle in the human oocyte[J]. Fertil Steril，1990，54 (1)：102-108.

[7] Ghetler Y，Yavin S，Shalgi R，et al. The effect of chilling on membranelipid phase transition in human oocytes and zygotes[J]. Hum Reprod，2005，20 (12)：3385-3389.

[8] Liebermann J，Dietl J，Vanderzwalmen P，et al. Recent developments in human oocyte，embryo and blastocyst vitrification：where are we now[J]. Reprod Biomed Online， 2003， 7 (6)：623-633.

[9] Lane M，Schoolcraft WB，Gardner DK. Vitrification of mouseand human blastocysts using a novel cryoloop container-lesstechnique [J]. Fertil Steril，1999，72 (6)：1073-1078.

[10] Kuwayama M，Vajta G，Kato O，et al. Highly efficient vitrification method for cryopreservation of human oocytes[J]. Reprod Biomed Online，2005，11 (3)：300-308.

[11] Lucena E，Bernal DP，Lucena C，et al．Successful ongoing pregnanciesaftervitrification ofooeytes[J]．Fertil Steril，2006， 85 (1)：108-11l.

[12] Hong SW，Chung HM．Lim JM，et al. Impmved human oocyte development after vitrification：a comparison of thawing methods [J]．Fertil Steril，1999，72 (1)：142-146．

[13] lsaehenko V，Montag M，Isaehenko E，et al．Aseptic vitrification of human germinal vesicle oocytes using dimethyl sulfoxide as aeryoproteetanL[J]. Fertil Steril，2006，85 (3)： 741-747.

[14] Rienzi L，Martinez F，Ubaidi F，et al．Polseope analysis of meiotic spindle chases in living metaphase Ⅱ human oocytesduring the freezing and thawing procedure[J].Hum Reprod，2004，19 (3)：655-659.

[15] Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos：the Cryotop method[J]. Theriogenology， 2007，67 (1)：73-80.

[16] Tedder RS，Zuckerman MA，Goldstone AH，et al. Hepatitis B transmission from contaminated cryopreservation tank[J]. Lancet，1995，346 (8968)：137-140.

[17] Bielanski A，Nadin-Davis S，Sapp T，et al. Viral contamination of embryos cryopreserved in liquid nitrogen[J]. Cryobiology， 2000，40 (2)：110-116.

冷冻载体及蔗糖浓度对复苏卵母细胞存活率的影响

The effects of the different combinations of freezing carrier and sucrose concentration on the oocyte survival after thawing

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