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具有识别发育能力良好的体外受精囊胚潜力的新策略。

Novel strategy with potential to identify developmentally competent IVF blastocysts.

作者信息

Jones Gayle M, Cram David S, Song Bi, Kokkali Georgia, Pantos Kostas, Trounson Alan O

机构信息

Monash Immunology and Stem Cell Laboratories (MISCL), Monash University, Level 3-STRIP Building 75, Wellington Road, Clayton, Victoria 3800, Australia.

出版信息

Hum Reprod. 2008 Aug;23(8):1748-59. doi: 10.1093/humrep/den123. Epub 2008 May 13.

Abstract

BACKGROUND

Currently there are no markers fully predictive of developmental competence of human IVF embryos. The present study investigated a novel strategy involving blastocyst biopsy and DNA fingerprinting to link developmental competence with gene expression patterns.

METHODS

Patient's blastocysts were biopsied to remove 8-20 trophectoderm (TE) cells for molecular analysis prior to transfer. Biopsy samples were amplified and gene expression was evaluated using microarrays. Sibling TE biopsies and cells from resulting offspring were subjected to DNA fingerprinting to identify which blastocyst(s) in the transfer cohort developed to term.

RESULTS

Blastocyst biopsy did not appear to impair developmental competence. Comparative microarray analysis of cDNA from pooled 'viable' and 'non-viable' TE samples identified over 7000 transcripts expressed exclusively in 'viable' blastocysts. The most significant of these included transcripts involved in cell adhesion and cell communication, key processes that have been associated with mammalian implantation. DNA fingerprinting of three cohorts of sibling blastocysts identified those blastocyst(s) that produced term pregnancies.

CONCLUSIONS

The combination of blastocyst biopsy, microarray gene expression profiling and DNA fingerprinting is a powerful tool to identify diagnostic markers of competence to develop to term. This strategy may be used to develop a rapid diagnostic assay or for refining existing criteria for the selection of the single most viable blastocyst among a cohort developing in vitro.

摘要

背景

目前尚无能够完全预测人类体外受精(IVF)胚胎发育能力的标志物。本研究调查了一种新策略,该策略涉及囊胚活检和DNA指纹识别,以将发育能力与基因表达模式联系起来。

方法

在移植前,对患者的囊胚进行活检,去除8 - 20个滋养外胚层(TE)细胞用于分子分析。对活检样本进行扩增,并使用微阵列评估基因表达。对同胞TE活检样本和由此产生的后代的细胞进行DNA指纹识别,以确定移植队列中的哪些囊胚发育至足月。

结果

囊胚活检似乎并未损害发育能力。对汇集的“存活”和“非存活”TE样本的cDNA进行比较微阵列分析,鉴定出7000多个仅在“存活”囊胚中表达的转录本。其中最显著的包括参与细胞黏附和细胞通讯的转录本,这些关键过程与哺乳动物着床有关。对三组同胞囊胚进行DNA指纹识别,确定了那些产生足月妊娠的囊胚。

结论

囊胚活检、微阵列基因表达谱分析和DNA指纹识别相结合是一种强大的工具,可用于识别发育至足月能力的诊断标志物。该策略可用于开发快速诊断检测方法,或用于完善现有标准,以在体外发育的一组囊胚中选择最具活力的单个囊胚。

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