Manor D, Kol S, Lewit N, Lightman A, Stein D, Pillar M, Itskovitz-Eldor J
Department of Obstetrics and Gynecology, Rambam Medical Center and Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
Hum Reprod. 1996 Nov;11(11):2502-6. doi: 10.1093/oxfordjournals.humrep.a019148.
Pronuclei formation is routinely assessed 16-20 h after oocyte insemination in in-vitro fertilization (IVF). Occasionally, the pronuclei disappear before this time, rendering them as 'undocumented'. Since the number of pronuclei detected is used to distinguish normal from abnormal embryos in the context of ploidy, the diploidy of undocumented embryos is questionable, and therefore they are routinely discarded. The introduction of fluorescent in-situ hybridization (FISH) technology allows the assessment of ploidy status in undocumented embryos that continue to cleave to form blostomeres. In this study, we used FISH to analyse the chromosomal status of 23 undocumented embryos obtained from 10 patients. Biopsied blastomeres were fixed and probed for five chromosomes (X, Y, 13, 18, 21). Diploidy was confirmed in 13 (57%) embryos while the remaining 10 embryos displayed various chromosomal anomalies. Six of the diploid embryos were transferred subsequently to the patients. One ongoing pregnancy was achieved following transfer of an undocumented, analysed embryo, which was already cleaved when assessed 20 h after insemination. We suggest that accelerated dismantling of the pronuclear membrane and subsequent cleavage do not necessarily indicate abnormal chromosomal content and may result in normal pregnancy. In a patient with a small number of embryos, FISH may be used to ascertain diploidy of undocumented embryos, thereby increasing the number of available embryos for transfer.
在体外受精(IVF)中,通常在卵母细胞受精后16 - 20小时评估原核形成情况。偶尔,原核会在此之前消失,使其成为“无记录的”。由于在倍性背景下,检测到的原核数量用于区分正常胚胎和异常胚胎,无记录胚胎的二倍体状态存在疑问,因此它们通常会被丢弃。荧光原位杂交(FISH)技术的引入使得能够评估继续分裂形成卵裂球的无记录胚胎的倍性状态。在本研究中,我们使用FISH分析了从10名患者获得的23个无记录胚胎的染色体状态。对活检的卵裂球进行固定,并针对五条染色体(X、Y、13、18、21)进行探针检测。在13个(57%)胚胎中确认了二倍体状态,而其余10个胚胎显示出各种染色体异常。随后将6个二倍体胚胎移植给了患者。在移植了一个无记录的、经分析的胚胎后实现了一例持续妊娠,该胚胎在受精后20小时评估时已经发生了分裂。我们认为,原核膜的加速解体和随后的分裂不一定表明染色体内容异常,并且可能导致正常妊娠。对于胚胎数量较少的患者,FISH可用于确定无记录胚胎的二倍体状态,从而增加可用于移植的胚胎数量。