The aim of this study is to investigate the effect of durations on nuclear maturation in vitro of feline oocytes. Following the in vitro maturation(IVM) in Medium 199 added with FSH and *** oocytes were treated for 28...
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The aim of this study is to investigate the effect of durations on nuclear maturation in vitro of feline oocytes. Following the in vitro maturation(IVM) in Medium 199 added with FSH and *** oocytes were treated for 28h.32h and 36h *** vitro maturated oocytes were subjected to artifical activation in 10%*** activated oocytes were cultured 6 *** rate of activated oocytes was evaluated on Day *** were stained with Hoechest 33342 on Day 6 to observe the potential of activated *** the same time,the matured oocytes were fixed in 3.7% paraformaldehyde,stained in FITC-LCA and *** observed the distribution of cortical granules(CGs) to evaluate the cytoplasmic *** cleave rate of activated oocytes in the 32h group was significantly higher than the 28h group (60.42%vs 43.16%.0.01
本试验以三种不同注射方式即精子注射、假性注射和不注射结合三种不同人工激活方法即注射CaCl2溶液(1.8p1，30mM)、离子霉素(15μmol/L，40min)和电脉冲(场强0.4KV/cm、脉冲时程90μs、一次脉冲)处理猪卵母细胞，以至少形成一个原核或发生卵裂计为激活，形成雌雄原核计为受精，在无精子注射的情况下至少形成两个原核或发生卵裂计为孤雌发育，统计分析不同激活方法对ICSI效果的影响。(1)机械刺激：研究发现假性注射+未激活处理组卵母细胞的激活率虽然高丁无机械刺激+无激活处理对照组((12.8％～13.3％)vs(8.3％～8.9％))，但明显低于假性注射+人工激活处理组((12.8％～13.3％)vs(18.3％～64.3％))，结果表明，单纯注射性机械刺激对猪卵母细胞的激活效果较差，为提高卵母细胞激活率，有必要进行人工激活;(2)人工激活：精子注射卵母细胞经CaCl2、离子霉素和电脉冲激活处理后，其激活率无显著性差异(69.4％vs 67.3％vs 70.4％)，CaCl2处理组的受精率显著高于离子霉素处理组(46.4％vs 31.2％)(P<0.05)，并与电激活处理组无显著性差异(46.4％ vs 38.8％)(P>0.05)。但是，在假性注射组(5.9％ vs 45.3％，51.6％)和对照组(4.5％ vs 38.8％，46.5％)中，CaCl2处理组的孤雌发育率极显著地低于离子霉素和电激活处理组(P<0.01);(3)CaCl2处理斤ICSI卵母细胞发育情况：CaCl2处理组的卵裂率(48.1％ vs 17.0％)(P<0.01)囊胚总细胞数(30.2vs 26.3)(P<0.05)显著高于未经CaCl2活化的对照组。
结果表明ICSI卵母细胞经CaCl2溶液激活处理后，能够取得较好的ICSI效果，而不显著增加其孤雌发育率。
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