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Positive effects of Forskolin (stimulator of lipolysis) treatment on cryosurvival of in vitro matured porcine oocytes

机译:佛司可林(脂解刺激物)治疗对体外成熟猪卵母细胞冷冻存活的积极影响

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In order to examine its effect on oocyte lipid content and cryosurvival, Forskolin was added to the medium for in vitro maturation of porcine oocytes. Treatments were control (IVM without Forskolin during the 42 h incubation period), addition of 10 mu M Forskolin for the entire 42 h (0-42) and addition of 10 mu M Forskolin between 24 and 42 h only (24-42). In Experiment 1, treatments did not differ significantly in cleavage rate, but the blastocyst formation rate was lower in the 0-42 group than for control and 24-42 group oocytes (17, 32 and 40%, respectively; P < 0.05). It was shown in Experiment 2 that Forskolin treatment from 0-42 h and from 24-42 h significantly reduced lipid content of oocytes compared to that of control cells (65 and 99 vs. 140 mu m(2) intensity of fluorescence, respectively; P < 0.05). In Experiment 3, the percentage of oocyte survival after cryopreservation and thawing was significantly higher in both Forskolin treatment groups than in control oocytes (72% for 0-42, 65% for 24-42 and 52% for control; P < 0.05). However, Forskolin treatment did not increase cleavage rates of vitrified in vitro matured porcine oocytes (Control group 28%, 0-42 h group 0%, 24-42 h group 26.67%). Addition of Forskolin affected the nuclear maturation of porcine oocytes. The percentage of PBE (polar body extrusion) were significantly reduced in the 0-42 h group (0-42 h group 42.00 +/- 2.08 vs. Control group 79.70 +/- 2.82 and 24-42 h group 70.60 +/- 2.83; P < 0.05). The 24-42 h group showed similar nuclear status to that of the Control group. We propose that delipation engendered by incubation with 10 mu M Forskolin during 24-42 hours of maturation increased cryosurvival of in vitro-maturated porcine oocytes and that attendant chemical lipolysis did not impair their further development as it may have done in oocytes incubated with Forskolin for the full 42 h
机译:为了检查其对卵母细胞脂质含量和冷冻存活的影响,将福司可林添加到用于猪卵母细胞体外成熟的培养基中。对照是治疗(在42 h的孵育期内不使用Forskolin的IVM),在整个42 h(0-42)内添加10μMForskolin,仅在24至42 h之间(24-42)添加10μMForskolin。在实验1中,治疗的卵裂率没有显着差异,但是0-42组的胚泡形成率低于对照组和24-42组卵母细胞(分别为17%,32%和40%; P <0.05)。在实验2中显示,从0-42小时和从24-42小时开始的佛司可林处理与对照细胞相比,卵母细胞的脂质含量显着降低(分别为65和99 vs. 140μm(2)荧光强度; P <0.05)。在实验3中,两个Forskolin治疗组的冷冻保存和解冻后的卵母细胞存活率显着高于对照组卵母细胞(0-42为72%,24-42为65%,对照组为52%; P <0.05)。然而,福斯高林治疗并没有增加玻璃化的体外成熟猪卵母细胞的裂解率(对照组28%,0-42h组0%,24-42h组26.67%)。佛司可林的添加影响了猪卵母细胞的核成熟。 0-42 h组(0-42 h组42.00 +/- 2.08与对照组79.70 +/- 2.82和24-42 h组70.60 +/- 2.83相比,PBE(极体挤压)的百分比显着降低; P <0.05)。 24-42 h组显示出与对照组相似的核状态。我们建议,在成熟的24-42小时内与10μMForskolin孵育引起的脱脂可增加体外成熟的猪卵母细胞的冷冻存活,并且伴随的化学脂解作用不会损害它们的进一步发育,因为在与Forskolin孵育的卵母细胞中整整42小时

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