超短精卵共孵育时间对小鼠体外受精的影响

江楠* 王玉洁 徐营 刘霞 蔡文伟 常永富 华科 

解剖学报 ›› 2013, Vol. 44 ›› Issue (2) : 279-283.

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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

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 
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 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
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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。

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.

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

Key words

Maleic dialdehyde / Oxidative stress / Ultrashort / In vitro fertilization / Embryo culture / Assisted reproductive technique / Mouse

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导出引用
江楠* 王玉洁 徐营 刘霞 蔡文伟 常永富 华科 . 超短精卵共孵育时间对小鼠体外受精的影响[J]. 解剖学报. 2013, 44(2): 279-283 https://doi.org/10.3969/j.issn.0529-1356.2013.02.026
JIANG Nan* WANG Yu-jie XU Ying LIU Xia CAI Wen-wei CHANG Yong-fu HUA Ke .  Effects of ultrashort gamete co-incubation time on mouse in vitro fertilization[J]. Acta Anatomica Sinica. 2013, 44(2): 279-283 https://doi.org/10.3969/j.issn.0529-1356.2013.02.026

参考文献

[1]    Plessis S, Makker K, Desai N,et al. Impact of oxidative stress on IVF[J]. Obstet Gynecol, 2008, 3(4): 539-554.

[2]Gianaroli L, Magli M, Ferraretti A, et al. Reducing the time of sperm-oocyte interaction in human in vitro fertilization improves the implantation rate[J]. Hum Reprod, 1996, 11(1): 166-171.

[3] Zhou Y, Huang XF, Lin JJ, et al. Impact on embryo quality and clinical outcome of reducing the time of sperm-oocyte interaction in human in vitro fertilization[J]. Chinese Journal of Obstetrics and Gynecology, 2003, 38(6): 373-374. (in Chinese)
周颖,黄学锋,林金菊,等. 缩短人精卵共孵育时间对体外受精\|胚胎移植中胚胎质量与临床结果的影响[J]. 中华妇产科杂志, 2003, 38(6): 373-374.
[4]Enkhmaa D, Kasai K, Hoshi K. Long-time exposure of mouse embryos to the sperm produces high levels of reactive oxygen species in culture medium and relates to poor embryo development[J]. Reprod Domest Anim, 2009, 44(4): 634-637.
[5]Bungum M,Bungum L, Humaidan P. A prospective study, using sibling oocytes, examining the effect of 30 seconds versus 90 minutes gamete co-incubation in IVF[J]. Hum Reprod, 2006, 21(2): 518-523.
[6]Nagy A, Gertsenstein M, Vintersten K, et al. Manipulating the Mouse Embryo: A Laboratory Manual [M]. 3rd edition. New York:Cold Spring Harbor Laboratory Press, 2003:411-434.
[7]Wassarman P. The biology and chemistry of fertilization[J]. Science, 1987, 235(4788): 553-560.
[8] Li T, Ren Z, Zhang MF, et al. Effects of ultrashort gamete co-incubation time on rate of polyspermy of human in vitro fertilization[J]. Journal of Sun Yet-Sen University(Medical Science), 2007, 28(Suppl 3): 67-69. (in Chinese)
李涛, 任姿, 张敏芳, 等. 超短时受精对人类配子体外多精受精发生率的影响[J]. 中山大学学报(医学科学版), 2007, 28(Suppl 3): 67-69.
[9]Alminana C, Gil M, Cuello C, et al. Effects of ultrashort gamete co-incubation time on porcine in vitro fertilization[J]. Anim Reprod Sci, 2008, 106(3-4): 393-401.
[10] Jiang N, Wang QZH. Effects of reducing gamete co-incubation time on outcome of human in vitro fertilization[J]. Chinese Journal of Birth Health and Heredity, 2011, 19(3): 7-8. (in Chinese)
江楠, 王启钊. 缩短精卵共孵育时间对人类体外受精结局的影响[J]. 中国优生与遗传杂志, 2011, 19(3): 7-8.
[11]Mahfouz R, Sharma R, Thiyagarajan A, et al. Semen characteristics and sperm DNA fragmentation in infertile men with low and high levels of seminal reactive oxygen species[J]. Fertil Steril, 2010, 94(6): 2141-2146.
[12] Li H, Yang XN, Zhao M, et al. Relationship between sperm DNA integrity and reactive oxygen species and uric acid in infertile men[J]. Chinese Journal of Family Planning, 2010, (3): 149-151. (in Chinese)
李红,杨学农,赵萌,等. 不育男性的精子DNA完整性与活性氧及尿酸的关系[J]. 中国计划生育学杂志, 2010, (3): 149-151. 
[13]Zini A, Boman J, Belzile E, et al. Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis[J]. Hum Reprod, 2008, 23(12): 2663-2668.
[14]Gil-Villa A, Cardona-Maya W, Agarwal A, et al. Assessment of sperm factors possibly involved in early recurrent pregnancy loss[J]. Fertil Steril, 2010, 94(4): 1465-1472. 

基金

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


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