褪黑素在体外成熟培养技术中对人类早期胚胎发育影响的探究
发布时间：2018-07-22 20:59
【摘要】：研究背景与常规IVF方案相比,人体外未成熟卵母细胞成熟技术(IVM)仍有很大的提升空间。褪黑素作为一种强效广谱抗氧化剂,可以清除体外培养条件下过多的氧自由基(ROS),从而抑制氧化应激对卵母细胞及胚胎的损伤。褪黑素添加于IVM培养液中是否可以促进IVM技术中未成熟卵母细胞成熟及发育潜能,从而作为改进IVM技术方案的突破点?本研究在人胚胎培养液中加入不同浓度褪黑素,探索对IVM技术中产生的人类早期胚胎的影响,旨在为IVM技术的改进提供理论依据。方法从2015年3月到2015年12月,共收集安徽医科大学第一附属医院生殖医学中心ICSI治疗周期患者未成熟卵母细胞353枚,患者年龄在35岁以下列入研究对象。人未成熟卵母细胞在含有10-5褪黑素的IVM培养液中培养至MⅡ期,培养成熟的卵母细胞行ICSI后,移至含有五组不同浓度褪黑素(0、10-11、10-9、10-7、10-5M)的胚胎培养液中继续培养,观察胚胎后续发育情况。挑选优质囊胚行array CGH检测,以评估褪黑素添加于IVM培养液及胚胎培养液,对囊胚安全性的影响。结果经过24~26 h的体外培养,共获得290枚成熟卵母细胞,用于评估褪黑素对胚胎后续发育的影响。胚胎卵裂率、优质囊胚率在五组之间均无统计学差异。囊胚率在4种褪黑素处理组中与对照组无显著差异,但是囊胚率在10-9M组最高,在10-5M组最低,10-9M组囊胚率显著高于10-5M组(47.1%vs 14.7%),差异具有统计学意义(P=0.002,c2=9.500)。在GV期和MⅠ期亚组中,卵裂率、囊胚率、优质囊胚率在不同浓度褪黑素处理的五组间均无显著差异。GV期亚组中。囊胚率在0M组最高(42.9%),在10-5M组最低(13.6%);优质囊胚率在0M组最高(19.0%),在10-5M组最低(4.5%)。在MⅠ期亚组中,囊胚率在10-9M组最高(64.0%),而在10-5M组最低(16.7%);优质囊胚率在10-9M组最高(24.0%),而在10-7M组最低(8.0%),本研究中共培养获得囊胚75枚,优质囊胚33枚。CGH的检测结果为:胚胎培养液褪黑素处理组的非整倍体率(12.5%,2/16)略低于对照组(16.7%,1/6),但是两组间无统计学差异。结论:(1)IVM培养液中加入褪黑素的前提下,在胚胎培养液中加入褪黑素,对人类早期胚胎的发育作用不大,低浓度褪黑素(0、10-11、10-9M)有增加囊胚形成率的倾向,但是高浓度褪黑素(10-7M、10-5M)会不利于胚胎的发育。(2)褪黑素添加在胚胎培养液中,可移植优质囊胚的非整倍体率没有增加,在一定程度上表明褪黑素用于人类胚胎培养是可行的。
[Abstract]:Background compared with conventional IVF protocols, IVM still has much room for improvement. Melatonin, as a powerful broad-spectrum antioxidant, can scavenge excessive oxygen free radicals (Ros) in vitro and inhibit oxidative stress damage to oocytes and embryos. Can melatonin be added to IVM medium to promote immature oocytes maturation and development potential in IVM technology as a breakthrough point to improve IVM technology? In this study, different concentrations of melatonin were added to the medium of human embryo culture to explore the effect of melatonin on the human early embryos produced in IVM technology, in order to provide theoretical basis for the improvement of IVM technology. Methods from March 2015 to December 2015, 353 immature oocytes were collected from ICSI patients in the Center of Reproductive Medicine, the first affiliated Hospital of Anhui Medical University. The patients, aged under 35 years, were included in the study. Human immature oocytes were cultured in IVM medium containing 10-5 melatonin to phase M 鈪，
本文编号：2138559
[Abstract]:Background compared with conventional IVF protocols, IVM still has much room for improvement. Melatonin, as a powerful broad-spectrum antioxidant, can scavenge excessive oxygen free radicals (Ros) in vitro and inhibit oxidative stress damage to oocytes and embryos. Can melatonin be added to IVM medium to promote immature oocytes maturation and development potential in IVM technology as a breakthrough point to improve IVM technology? In this study, different concentrations of melatonin were added to the medium of human embryo culture to explore the effect of melatonin on the human early embryos produced in IVM technology, in order to provide theoretical basis for the improvement of IVM technology. Methods from March 2015 to December 2015, 353 immature oocytes were collected from ICSI patients in the Center of Reproductive Medicine, the first affiliated Hospital of Anhui Medical University. The patients, aged under 35 years, were included in the study. Human immature oocytes were cultured in IVM medium containing 10-5 melatonin to phase M 鈪，
本文编号：2138559
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