精子孵育时间及培养液成分对小鼠体外受精和胚胎发育的影响

目的探讨精子孵育时间及培养液成分对小鼠体外受精和胚胎发育的影响,为人类辅助生殖技术完善质量控制体系创建可靠的平台。方法将精子获能时间分为3组,分别为0.5、1和2h,用HTF＋10%SSS和HTF＋10%FCS两种获能受精液,ICR雄鼠附睾尾的精子与成熟卵母细胞进行体外受精（IVF）。受精卵分别用IVC-ONE＋10%SSS、IVC-ONE＋10%FCS、HTF＋10%FCS和HTF＋10%SSS4种培养液对体外受精胚胎进行培养。结果结果获能时间为0.5h时,卵母细胞的受精率显著高于其他组,差异有高度统计学意义（P〈0.01）;获能时间若超过1h,受精率明显下降。胚胎用IVC-ONE＋10%FCS培养液进行培养时囊胚率和囊胚孵出率（89%和40%）显著高于IVC-ONE＋10%SSS（51%和29%）、HTF＋10%FCS（11%和2%）和HTF＋10%SSS（11%和0%）,差异有高度统计学意义（P〈0.01）。结论小鼠体外受精时，宜选用HTF受精液，并获能0．5h；小鼠体外受精卵进行培养，以含10％FCS的IVC＋ONE为胚胎培养液，可获得较高的囊胚率，葡萄糖并不是小鼠胚胎形成囊胚所必需的；在含磷酸盐的培养液中EDTA不能有效克服ICR2-cell的体外发育阻滞现象。

Effect of Sperm Capacitation Time and Culture Medium Components on in vitro Fertilization and Embryo Development of Mice

Objective The experiments were carried out to explore the effect of sperm capacitation time and culture medium components on in vitro fertilization （IVF） and culture （IVC） of early mice em- bryos in vitro, and to develop an improved early embryos culture system for mice. Methods Sperm ca- pacitation time was divided into three groups, 0.5h（group I ）, 1h（group Ⅱ） and 2h（group Ⅲ）. Sperm de- rived from epididymal tail were added to mature oocytes for IVF in different culture medium i.e. HTF＋ 10%SSS and HTF ＋10%FCS. The embryos were cultured in IVC-ONE＋IO%SSS, IVC-ONE＋10%FCS, HTF＋10%FCS and HTF＋10%SSS. Results The IVF rate in group I was significantly higher than that in group Ⅱ and Ⅲ （/9〈0.01） . If the Capacitation time was more than lh, the IVF rate would significantly decrease. The blastocyst rate and hatched blastocyst rate cultured in IVC-ONE ＋ 10%FCS medium were 89% and 40%, which was significantly higher than in IVC-ONE ＋ 10%SSS （51% and 29%） ,HTF＋10% FCS （11%和 2%）and HTF＋10%SSS （11% and 0% ）. Conclusion When HTF is used as fertilization medium and capacitation time is 0.5h the IVF rate is significantly higher than that in the other groups in which other culture mediums are used with different capacitation time. The blastocyst formation rate in IVC-ONE ＋ 10%FCS is higher than in other culture mediums. Glucose is not necessary for mouse blastocyst formation. Containing phosphate in the culture medium EDTA can not overcome the 2-cell block effectively development in vitro.