Unknown,Transcriptomics,Genomics,Proteomics

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Molecular signature in human cumulus cells related to embryonic developmental potential

ABSTRACT: The objective of the present study was to investigate gene expression profiles of human Cumulus cells isolated from oocytes at metaphase II stage, under controlled ovarian stimulation (COS) cycle and to evaluate the relationship of embryo quality with molecular signatures in Cumulus cells. This study has been performed by microarray analysis in order to identify potential biomarkers related to oocyte developmental potential. For each patient, the CCs were classified into two groups (good quality embryo group and bad quality embryo group) and pooled. From the total of 14 samples, total RNA was purified. After removal of rRNA, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3’ bias utilizing a random priming method. The amplified and labelled cRNA were pooled as good quality and bad quality group followed by hybridization, scanning and data analysis.Three independent repetitions were carried out, thus a total of 6 chips were employed in this study. the objective of the present study was to investigate gene expression profiles of human CCs isolated from oocytes at metaphase II stage, under controlled ovarian stimulation (COS) cycle and to evaluate the relation of following embryo quality to molecular signatures in CCs.

ORGANISM(S): Homo sapiens

SUBMITTER: Dawei Chen

PROVIDER: E-GEOD-54135 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Purpose: The goal of this study was to identify differentially expressed genes and pathways between the cumulus of compact/unstimulated cumulus-oocyte-complex (COC) and the cumulus of expanded/stimulated COC. Methods: mRNA profiles of Compact/unstimulated cumulus cells (CCs) from germinal vesicle (GV) COC obtained from two patients undergoing unstimulated IVM procedure and expanded/stimulated CCs from metaphase 2 (MII) COC obtained from three patients undergoing IVF/ICSI were generated by deep sequencing using Illumina HiSeq 2000. The sequence reads that passed quality filters were mapped to the human genome (hg19) using Tophat software. Differential expression analysis was done using DESeq bioconductor package. qRT–PCR validation was performed using SYBR Green assays. Results: A total of 40-80 million sequence reads per sample were mapped to the human genome (hg19). A total of 1746 differentially expressed genes between compact and expanded CCs with fold change > 2, and adjusted p value < 0.05 were identified. Gene ontology analysis of differentially expressed genes revealed a number of cellular processes regulated during the periovulatory interval including cellular movement, inflammatory response, immune cell trafficking, tissue development, lipid metabolism, tissue morphology, DNA replication and cell cycle. A total of 116 of the differentially expressed genes were annotated as long non-coding RNAs, 10 of them coded from introns of genes known to be involved in granulosa cell processes suggesting that unique non coding RNA transcripts may contribute to the regulation of cumulus expansion and oocyte maturation. Results were validated using qRT-PCR. Conclusions: Using global transcriptome sequencing we identified new important genes and non coding RNAs involved in COC maturation and cumulus expansion, which may contribute to improve the process of in vitro maturation of immature oocytes utilized in IVM cycles mRNA profiles of Compact/unstimulated cumulus cells (CCs) from germinal vesicle (GV) COC obtained from two patients undergoing unstimulated IVM procedure and expanded/stimulated CCs from metaphase 2 (MII) COC obtained from three patients undergoing IVF/ICSI were generated by deep sequencing using Illumina HiSeq 2000.

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Molecular signature in human cumulus cells related to embryonic developmental potential

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