兔卵母细胞的超数排卵与孤雌激活

Superovulation and parthenogenetic activation in rabbits

目的:探索兔卵母细胞的最佳超排方法及孤雌激活的条件,为人-兔异种核移植提供充足的兔卵母细胞. 方法:采用不同剂量的垂体促卵泡素(FSH)对青年兔和经产兔进行超数排卵,获得兔卵母细胞后,以不同强度的直流电脉冲对兔卵母细胞进行孤雌激活. 结果:在排卵总数方面,3个激素组均高于对照组(P＜0.05),且FSH 25 U激素组(31.80±3.27)与FSH 80 U+聚乙烯吡咯酮(PVP)激素组(31.83±3.06)均高于FSH 15 U激素组(15.00±1.58),经产兔优于青年兔(P<0.05);在200 V/mm组孤雌激活兔卵母细胞的卵裂率(85.59%)和4～8细胞期发育率(21.19%)最高,与240 V/mm组(71.43%, 10.48%)之间存在统计学差异(P＜0.05). 结论:FSH 25 U及FSH 80 U+PVP方案均能产生最佳超排效果,经产兔优于青年兔;电场强度200 V/mm,脉宽40 μs,2次脉冲、间隔1 s为兔成熟卵母细胞的较理想的激活条件.

AIM： To explore optimal superovulating methods and parthenogenetic activation conditions in rabbits. METHODS： Virginal and multiparous rabbits were superovulated by follicle-stimulating hormone （ FSH ） at different concentrations. After being obtained, the mature oocytes were activated by electric current with different strength. RESULTS： There were significant differences in the ovulating total numbers between hormone groups and control group （P 〈 0.0.5 ） and between the group with FSH 1.5 U and the other hormone groups [ （ 15.00 ± 1.58） vs （31.83 ± 3.06）, （ 31. 80 ± 3.27 ）, P 〈 0.05 ]. So was the difference between virginal rabbits and the multiparous rabbits. The rates of merogenesis （ 85.59% ） and the rates of development to 4 - 8 cell stage（21. 19% ） of rabbit oocytes were the highest in the group with 200 V/mm direct current pulses treatment among the four groups. CONCLUSION： The superovulation efficiency in rabbits with FSH （2.5 U ） or FSH （80 U） ＋ PVP treatment is optimized and the superovulation efficiency is better in multiparous rabbits than that in virginal rabbits. The comparatively suitable oocyte activation condition is that of 200 V/mm, 40 us and 2 times of direct current pulses with a 1 s of interval.