兔卵母细胞超数排卵及孤雌激活.docVIP

兔卵母细胞超数排卵及孤雌激活 　 作者：王春雨，鲁进，武浩，魏欣，赵甫涛，贾战生 【关键词】 卵母细胞 Superovulation and parthenogenetic activation in rabbits 　　【Abstract】 AIM: To explore optimal superovulating methods and parthenogenetic activation conditions in rabbits. METHODS: Virginal and multiparous rabbits were superovulated by folliclestimulating 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 (Plt;0.05) and between the group with FSH 15 U and the other hormone groups ［(15.00±1.58) vs (31.83±3.06), (31.80±3.27), Plt;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 (25 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 μs and 2 times of direct current pulses with a 1 s of interval. 　　【Keywords】 oocytes; parthenogenetic activation; rabbit 　　【摘要】 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（Plt;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. 　　【关键词】 卵母细胞；孤雌激活；兔 　　0引言 　　治疗性克隆（therapeutic cloning）是体细胞核移植技术和胚胎干细胞技术相结合的产物，将成为人类医疗史上革命性的技术. 由于人类卵母细胞来源非常有限，且受到法律、伦理等多种因素的限制，很难获得足够数量的人卵母细胞用于实验研究. 异种核移植技