解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (2 ): 279-283.doi: 10.3969/j.issn.0529-1356.2013.02.026

• 组织学胚胎学发育生物学 • 上一篇    下一篇

超短精卵共孵育时间对小鼠体外受精的影响

江楠1,3*  王玉洁2  徐营3  刘霞1  蔡文伟1  常永富1  华科1    

  1. 1.    嘉兴市妇幼保健院生殖医学中心，浙江 嘉兴 314050; 2. 嘉兴学院生物与化学工程学院生物工程系，浙江 嘉兴 314001;3. 嘉兴学院医学院组织学与胚胎学教研室，浙江 嘉兴 314001） 
  • 收稿日期:2012-08-23 修回日期:2012-09-20 出版日期:2013-04-06 发布日期:2013-04-06
  • 通讯作者: 江楠 E-mail:jn007_2001@hotmail.com
  • 基金资助:

    浙江省嘉兴市科技计划项目（NO.2009BY7011）;市自然科学基金资助项目;自然科学基金资助项目

 Effects of ultrashort gamete co-incubation time on mouse in vitro fertilization

JIANG Nan 1,3* WANG Yu-jie2 XU Ying3   LIU Xia1 CAI Wen-wei1 CHANG Yong-fu1 HUA Ke1    

  1. 1. Reproductive Medicine Center, Jiaxing Maternal and Child Health Hospital, Zhejiang Jiaxing 314050, China; 2. Department of Biological Engineering,School of Biological and Chemical Engineering, Jiaxing University, Zhejiang Jiaxing 314001, China; 3. Department of Histology and Embryology,School of Medicine, Jiaxing University, Zhejiang Jiaxing 314001, China
  • Received:2012-08-23 Revised:2012-09-20 Online:2013-04-06 Published:2013-04-06

摘要：

目的 探讨不同精卵共孵育时间对小鼠体外受精受精率、氧化应激产生及胚胎质量的影响，及小鼠体外受精最短的精卵共孵育时间。 方法 分别比较ICR小鼠精卵共孵育30s、2min、10min、05h、2h、4h及18h的体外受精的受精率及囊胚形成率，并对精卵共孵育0h、2h及18h培养液的丙二醛浓度进行检测。结果 各组获得卵细胞数分别为52、53、53、46、48、48及47，受精率分别为67.3%、81.1%、83.0%、87.0%、85.4%、81.3%及48.9%。精卵共孵育30s组的受精率显著低于0.5h及2h组（P ＜0.05），而18h组的受精率显著低于2min、10min、0.5h、2h及4h组（P ＜0.05）。2min组的受精率相比10min、0.5h、2h及4h组均无差异（P ＞0.05）。各组的囊胚形成率分别为82.9%、65.1%、70.5%、67.5%、75.6%、71.8%及65.2%，两两比较差异均无显著性（ P ＞0.05）。精卵共孵育0h、2h及18h丙二醛浓度检测的均值分别为（1.06±0.21）×10-3mol/L、（1.44±0.22）×10-3mol/L及（2.00±0.21）×10-3mol/L，各组间两两比较差异均有显著性（ P ＜0.05），丙二醛浓度随着精卵共孵育时间的缩短而降低。结论 小鼠体外受精精卵共孵育时间缩短至2min不会降低受精率，同时随着孵育时间的缩短，产生的氧化应激也随之减少。

关键词: 丙二醛 , 氧化应激 , 超短 , 体外受精 ,  胚胎培养 ,  辅助生殖技术 ,  小鼠

Abstract:

Objective To study the impact of gamete co-incubation time on fertilization rate, production of oxidative stress (OS) and embryo quality of mouse in vitro fertilization (IVF), and to investigate the shortest gamete co-incubation time of mouse IVF. Methods Oocytes of ICR mouse were co-incubated with sperm for 30 seconds, 2 minutes, 10 minutes , 0.5 hour, 2 hours, 4 hours and 18 hours, the fertilization rate and the blastocyst rate were analysed in each co-incubation group. In addition, the maleic dialdehyde(MDA) was assessed when gametes were co-incubated for 0 hour, 2 hours and 18 hours. Results Fifty-two, fifty-three, fifty- three, fourty-six, fourty-eight, fourty-eight and fourty-seven oocytes were inseminated with sperm in each group, and the fertilization rate were 67.3%, 81.1%, 83.0%, 87.0%, 85.4%, 81.3% and 48.9% respectively. The fertilization rate of 30 seconds group was significantly lower than 0.5 hour and 2 hours groups（ P ＜0.05）. Moreover, the fertilization rate of 18 hours group was significantly lower than 2 minutes, 10min, 0.5 hour, 2 hours and 4 hours groups（P ＜0.05）. No significant differences in fertilization rate were observed in 2 minutes, 10 minutes, 0.5 hour, 2 hours and 4 hours groups（P ＞0.05）. In addition, the blastocyst rate were 82.9%, 65.1%, 70.5%, 67.5%, 75.6%, 71.8% and 65.2%, and no significant differences were observed in each group（P ＞0.05）. The concentrations of MDA were
(1.06±0.21)×10-3 mol/L, (1.44±0.22)×10-3 mol/L and (2.00±0.21)×10-3 mol/L for 0 hour, 2 hours and 18 hours co-incubation, respectively. As the coincubation time was reduced, the MDA levels were also significantly reduced（P ＜0.05）. Conclusion Reducing the gamete co-incubation time to 2 minutes will not affect the fertilization rate of mouse IVF. As the co-incubation time is reduced, the production of OS is also reduced.

Key words: Maleic dialdehyde , Oxidative stress , Ultrashort , In vitro fertilization , Embryo culture , Assisted reproductive technique , Mouse