对体外受精和卵胞浆内单精子注射“受精失败”的人类卵母细胞进行细胞遗传学和荧光原位杂交染色体研究。

Cytogenetic and fluorescent in-situ hybridization chromosomal studies on in-vitro fertilized and intracytoplasmic sperm injected 'failed-fertilized' human oocytes.

作者信息

Wall M B, Marks K, Smith T A, Gearon C M, Muggleton-Harris A L

机构信息

Division of Obstetrics and Gynaecology, UMDS, St Thomas' Hospital, London, UK.

出版信息

Hum Reprod. 1996 Oct;11(10):2230-8. doi: 10.1093/oxfordjournals.humrep.a019082.

DOI:10.1093/oxfordjournals.humrep.a019082

PMID:8943535

Abstract

This study was undertaken to establish baseline data on the chromosomal status of 'failed-fertilized' oocytes derived from in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) procedures. A cytogenetic analysis was undertaken on 162 IVF and 51 ICSI oocytes. In all, 82.1% (133/162) of the IVF and 78.4% (40/51) of the ICSI oocytes had metaphase II (MII) plates, of which 50.4% of the IVF and 47.5% of the ICSI oocytes were analysed further. Chromosomes of the G-group (21-22) were identified with the majority of the anomalies. No overall significant difference in the aneuploidy rate was found for the IVF (37.3%) of ICSI (31.6%) oocytes, or with maternal age. However, chromosome anomalies, e.g. diploidy, fragmented and broken chromatids, single sperm and oocyte chromatids, were found in oocytes from IVF patients aged > 36 years and in the ICSI oocytes throughout the maternal age range (31-38 years). The status of the polar body chromatin indicated that there was no overall significant difference in the maturation of the IVF and ICSI oocytes. Evidence of successful sperm delivery was found in 72.5% (37/51) of the ICSI failed-fertilized oocytes. In this group there was a significant increase in the incidence of premature chromosome condensation: 19.6% (10/51) contained sperm chromosomes, 7.8% (4/51) had swollen sperm heads, and the remaining 45.0% had condensed sperm heads. The presence of both sperm and MII oocyte chromosomes was found in 19.6% (10/51) of the ICSI and 8.6% (14/162) of the IVF failed-fertilized oocytes. Specific fluorescent in-situ hybridization DNA probes were used to re-analyse the chromosomes of karyotyped 'failed-fertilized' IVF oocytes and, for the first time, applied to the karyotyped chromosomes of failed-fertilized ICSI oocytes. The hybridization efficiency was 86-95% for the centromere probe and 100% for probes 21 and 18.

摘要

本研究旨在建立关于体外受精（IVF）或卵胞浆内单精子注射（ICSI）过程中“受精失败”卵母细胞染色体状态的基线数据。对162个IVF卵母细胞和51个ICSI卵母细胞进行了细胞遗传学分析。总体而言，82.1%（133/162）的IVF卵母细胞和78.4%（40/51）的ICSI卵母细胞具有中期II（MII）板，其中50.4%的IVF卵母细胞和47.5%的ICSI卵母细胞被进一步分析。G组（21 - 22）染色体的异常最为常见。IVF（37.3%）和ICSI（31.6%）卵母细胞的非整倍体率以及与母亲年龄之间均未发现总体显著差异。然而，在年龄大于36岁的IVF患者的卵母细胞以及整个母亲年龄范围（31 - 38岁）的ICSI卵母细胞中发现了染色体异常，如二倍体、染色单体片段化和断裂、单个精子和卵母细胞染色单体。极体染色质状态表明IVF和ICSI卵母细胞的成熟度总体上没有显著差异。在72.5%（37/51）的ICSI受精失败卵母细胞中发现了成功授精的证据。在该组中，早熟染色体凝聚的发生率显著增加：19.6%（10/51）含有精子染色体，7.8%（4/51）精子头部肿胀，其余45.0%精子头部凝聚。在19.6%（10/51）的ICSI和8.6%（14/162）IVF受精失败卵母细胞中发现了精子和MII卵母细胞染色体共存的情况。使用特异性荧光原位杂交DNA探针重新分析了核型分析的“受精失败”IVF卵母细胞的染色体，并首次应用于核型分析的受精失败ICSI卵母细胞的染色体。着丝粒探针的杂交效率为86 - 95%，21号和18号探针的杂交效率为100%。