【摘要】：研究背景與常規(guī)IVF方案相比,人體外未成熟卵母細(xì)胞成熟技術(shù)(IVM)仍有很大的提升空間。褪黑素作為一種強(qiáng)效廣譜抗氧化劑,可以清除體外培養(yǎng)條件下過多的氧自由基(ROS),從而抑制氧化應(yīng)激對(duì)卵母細(xì)胞及胚胎的損傷。褪黑素添加于IVM培養(yǎng)液中是否可以促進(jìn)IVM技術(shù)中未成熟卵母細(xì)胞成熟及發(fā)育潛能,從而作為改進(jìn)IVM技術(shù)方案的突破點(diǎn)?本研究在人胚胎培養(yǎng)液中加入不同濃度褪黑素,探索對(duì)IVM技術(shù)中產(chǎn)生的人類早期胚胎的影響,旨在為IVM技術(shù)的改進(jìn)提供理論依據(jù)。方法從2015年3月到2015年12月,共收集安徽醫(yī)科大學(xué)第一附屬醫(yī)院生殖醫(yī)學(xué)中心ICSI治療周期患者未成熟卵母細(xì)胞353枚,患者年齡在35歲以下列入研究對(duì)象。人未成熟卵母細(xì)胞在含有10-5褪黑素的IVM培養(yǎng)液中培養(yǎng)至MⅡ期,培養(yǎng)成熟的卵母細(xì)胞行ICSI后,移至含有五組不同濃度褪黑素(0、10-11、10-9、10-7、10-5M)的胚胎培養(yǎng)液中繼續(xù)培養(yǎng),觀察胚胎后續(xù)發(fā)育情況。挑選優(yōu)質(zhì)囊胚行array CGH檢測(cè),以評(píng)估褪黑素添加于IVM培養(yǎng)液及胚胎培養(yǎng)液,對(duì)囊胚安全性的影響。結(jié)果經(jīng)過24~26 h的體外培養(yǎng),共獲得290枚成熟卵母細(xì)胞,用于評(píng)估褪黑素對(duì)胚胎后續(xù)發(fā)育的影響。胚胎卵裂率、優(yōu)質(zhì)囊胚率在五組之間均無統(tǒng)計(jì)學(xué)差異。囊胚率在4種褪黑素處理組中與對(duì)照組無顯著差異,但是囊胚率在10-9M組最高,在10-5M組最低,10-9M組囊胚率顯著高于10-5M組(47.1%vs 14.7%),差異具有統(tǒng)計(jì)學(xué)意義(P=0.002,c2=9.500)。在GV期和MⅠ期亞組中,卵裂率、囊胚率、優(yōu)質(zhì)囊胚率在不同濃度褪黑素處理的五組間均無顯著差異。GV期亞組中。囊胚率在0M組最高(42.9%),在10-5M組最低(13.6%);優(yōu)質(zhì)囊胚率在0M組最高(19.0%),在10-5M組最低(4.5%)。在MⅠ期亞組中,囊胚率在10-9M組最高(64.0%),而在10-5M組最低(16.7%);優(yōu)質(zhì)囊胚率在10-9M組最高(24.0%),而在10-7M組最低(8.0%),本研究中共培養(yǎng)獲得囊胚75枚,優(yōu)質(zhì)囊胚33枚。CGH的檢測(cè)結(jié)果為:胚胎培養(yǎng)液褪黑素處理組的非整倍體率(12.5%,2/16)略低于對(duì)照組(16.7%,1/6),但是兩組間無統(tǒng)計(jì)學(xué)差異。結(jié)論:(1)IVM培養(yǎng)液中加入褪黑素的前提下,在胚胎培養(yǎng)液中加入褪黑素,對(duì)人類早期胚胎的發(fā)育作用不大,低濃度褪黑素(0、10-11、10-9M)有增加囊胚形成率的傾向,但是高濃度褪黑素(10-7M、10-5M)會(huì)不利于胚胎的發(fā)育。(2)褪黑素添加在胚胎培養(yǎng)液中,可移植優(yōu)質(zhì)囊胚的非整倍體率沒有增加,在一定程度上表明褪黑素用于人類胚胎培養(yǎng)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，

本文編號(hào)：2138559