精子数量对小鼠体外受精与胚胎早期发育的影响

Effects of Sperm Counts on In Vitro Fertilization and Early Embryonic Development of Mice

【目的】研究不同精子数量对小鼠体外受精与胚胎早期发育的影响,通过比较分析进一步完善小鼠的辅助生殖技术。【方法】试验随机选用9周龄C57BL/6雄鼠附睾尾获取的精子放入HTF培养液,在二氧化碳培养箱中培养1 h后通过精子计数计算1、3、5、10、15、20μL精子溶液的精子数量,并吸取相应的精子溶液于4周龄C57BL/6雌鼠超排后获取的卵母细胞团中,体外培养24 h后计算体外受精率与2-细胞胚胎发育率。【结果】1、3、5、10、15、20μL精子溶液的精子数量分别为0.25(±0.23)×10^6、1.23(±0.31)×10^6、2.03(±0.35)×10^6、3.95(±0.33)×10^6、6.50(±0.76)×10^6、9.88(±0.52)×10^6个/mL;同时,小鼠体外受精率分别为88.92%、95.17%、94.29%、90.56%、85.89%、78.26%,其中2-细胞胚胎率分别为78.21%、81.75%、81.51%、77.42%、70.47%、50.76%。精子数量为1.23(±0.31)×10^6、2.03(±0.35)×10^6个/mL的体外受精率与2-细胞胚胎发育率较好,差异不显著,但极显著高于精子计数为6.50(±0.76)×10^6、9.88(±0.52)×10^6个/mL的情况。【结论】不同的精子数量对小鼠体外受精与胚胎早期发育影响有显著性差异,精子数量为1.23(±0.31)×10^6~2.03(±0.35)×10^6个/mL时能有效提高小鼠的体外受精率并促进胚胎的正常发育。

【Objective】The effects of different sperm counts on in vitro fertilization and early embryonic development of mice were studied,and the assisted reproductive technique of mice was further improved by comparative analysis.【Methods】In the test,sperms obtained from the epididymal tail of 9 weeks old C57BL/6 male mice were randomly selected and put into HTF culture medium,which cultured in carbon dioxide incubator for 1 hour.The sperm counts of semen solution of 1,3,5,10,15 and 20μL were calculated,the corresponding semen solution absorbed into the oocyte clusters which obtained after the superovulation of 4 weeks old C57BL/6 female mice,and the in vitro fertilization rate and the development rate of 2-cell embryos calculated after in vitro culture for 24 hours.【Results】Sperm counts of 1,3,5,10,15 and 20μL semen solution were 0.25(±0.23)×10^6,1.23(±0.31)×10^6,2.03(±0.35)×10^6,3.95(±0.33)×10^6,6.50(±0.76)×10^6,and 9.88(±0.52)×10^6(units/mL).At the same time,the in vitro fertilization rates of mice were 88.92%,95.17%,94.29%,90.56%,85.89%and 78.26%,respectively,and the development rates of 2-cell embryos were 78.21%,81.75%,81.51%,77.42%,70.47%and 50.76%,respectively.The in vitro fertilization rates and the 2-cell embryo development rates with sperm counts of 1.23(±0.31)×10^6 and 2.03(±0.35)×10^6(units/mL)were better than those of others,with no significant difference,but were extremely significantly higher than those with sperm counts of 6.50(±0.76)×10^6,9.88(±0.52)×10^6(units/mL).【Conclusions】Various sperm counts have significantly different effects on the in vitro fertilization and early embryonic development of mice.The appropriate sperm numbers were 1.23(±0.31)×10^6-2.03(±0.35)×10^6(units/mL),which can effectively improve the in vitro fertilization rates of mice and promote the normal development of embryos.