超短精卵共孵育时间对小鼠体外受精的影响
Effects of ultrashort gamete co-incubation time on mouse in vitro fertilization
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、05h、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)×10-3mol/L、(1.44±0.22)×10-3mol/L及(2.00±0.21)×10-3mol/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。
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 coincubation 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.
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
Maleic dialdehyde / Oxidative stress / Ultrashort / In vitro fertilization / Embryo culture / Assisted reproductive technique / Mouse
[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.
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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