Mélanie Hamel

Université de Montréal, Montréal, Quebec, Canada

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Publications (4)17.78 Total impact

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    ABSTRACT: Multiple pregnancy represents an important health risk to both mother and child in fertility treatment. To reduce a high twin rate, restriction to one embryo transfer is needed. Morphological evaluation methods for predicting embryo viability has significant limitations. Tight communication exists between the follicular cells (FCs) and the oocyte; therefore, developmental competence may be determined by markers expressed in the surrounding FCs. In this study, cells were recovered on a per-follicle basis by individual follicle puncture. Hybridization analysis using a custom-made complementary DNA microarray containing FC transcripts was performed. Genes expressed in FCs associated with good morphological transferred embryos were identified from follicles associated with a pregnancy outcome (pregnancy group) or no pregnancy (non-pregnancy group). Ten candidates from the Pregnancy group and three from the Non-pregnancy group were validated by quantitative RT-PCR. The best predictors associated with pregnancy were UDP-glucose pyrophosphorylase-2 and pleckstrin homology-like domain, family A, member 1. Genes assessment showed no significant candidate genes associated with non-pregnancy outcome, but GA-binding protein transcription factor beta1 showed a tendency to be potentially more expressed in the non-pregnancy group. These markers could be related to granulosa luteinization process and could be used to improve embryo selection for successful single embryo transfer.
    Molecular Human Reproduction 08/2010; 16(8):548-56. · 4.54 Impact Factor
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    ABSTRACT: Embryo selection efficiency in human IVF procedure is still suboptimal as shown by low pregnancy rates with single embryo transfer (SET). Bidirectional communication between the oocyte and follicular cells (FC) is essential to achieve developmental competence of the oocyte. Differences in the gene expression profile of FCs from follicles leading to pregnancy could provide useful markers of oocyte developmental competence. FCs were recovered by individual follicle puncture. FC expression levels of potential markers were assessed by Q-PCR with an intra-patient and an inter-patient analysis approach. Using gene expression, a predictive model of ongoing pregnancy was investigated. Using intra-patient analysis, four candidate genes, phosphoglycerate kinase 1 (PGK1), regulator of G-protein signalling 2 (RGS2), regulator of G-protein signalling 3 (RGS3) and cell division cycle 42 (CDC42) showed a difference between FCs from follicles leading to a pregnancy or developmental failure. The best predictors for ongoing pregnancy were PGK1 and RGS2. Additionally, inter-patient analysis revealed differences in FC expression for PGK1 and CDC42 between follicles leading to a transferred embryo with positive pregnancy results and those with negative results. Both inter-patient and intra-patient approaches must be taken into consideration to delineate gene expression variations in the context of follicular competence. A predictor model using biomarkers could improve the efficiency of predicting developmental competence of oocytes. These new approaches provide useful tools in the context of embryo selection and in the improvement of pregnancy rates with SET.
    Molecular Human Reproduction 09/2009; 16(2):87-96. · 4.54 Impact Factor
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    ABSTRACT: The development of an accurate method for selection of high-quality embryos is essential to achieve high pregnancy rates with single embryo transfer in human IVF. The developmental competence of the oocyte is acquired during follicle maturation and strong communication also exists between the follicular cells (FCs) and the oocytes; thus oocyte developmental competence may be determined by markers expressed in the surrounding FCs. From consenting patients (n = 40), FCs were recovered on a per follicle basis by individual follicle puncture. Hybridization analyses using a custom-made complementary DNA microarray containing granulosa/cumulus expressed sequence tags (ESTs) from subtracted libraries and an Affymetrix GeneChip were performed to identify specific genes expressed in follicles leading to a pregnancy. The selected candidate genes were validated by quantitative-PCR (Q-PCR). Subtractive libraries prepared from pooled samples representing pregnant versus non-pregnant patients produced 1694 ESTs. Hybridization data analysis discriminated 115 genes associated with competent follicles. Selected candidates were confirmed by Q-PCR: 3-beta-hydroxysteroid dehydrogenase 1 (P = 0.0078), Ferredoxin 1 (P = 0.0203), Serine (or cysteine) proteinase inhibitor clade E member 2 (P = 0.0499), Cytochrome P450 aromatase (P = 0.0359) and Cell division cycle 42 (P = 0.0396). Microarray technologies are useful to mine the transcriptome of FCs expressed in follicles associated with competent oocytes and could be used to improve embryo selection with the objective of successful single embryo transfer.
    Human Reproduction 06/2008; 23(5):1118-27. · 4.67 Impact Factor
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    ABSTRACT: Oocyte competence is the ability of the oocyte to complete maturation, undergo successful fertilization, and reach the blastocyst stage. Cumulus cells are indispensable for this process. Their removal significantly affects the blastocyst rates. Moreover, the properties and functions of cumulus cells are regulated by the oocyte. They also reflect the oocyte's degree of maturation. Our study was aimed at identifying markers of oocyte competence that are expressed in bovine cumulus cells. In a previous study in our laboratory, the blastocyst yield following FSH or phorbol myristate acetate (PMA) treatment was 45%%. Therefore, we tested four sets of conditions during the first 6 h of in vitro maturation (IVM): FSH (0.1 microg/ml), PMA (0.1 microM), FSH ++ PMA, and negative control. Extracts from each IVM treatment were hybridized against the same negative control on a microarray containing a partial library of differentially expressed transcripts in the cumulus of competent oocytes collected at 6 h after LH in vivo. Common positive clones between diffrentially treated cells were selected, and 15 candidates were validated by real-time PCR. Based on this, the main candidates expressed in cumulus cells and that could be valuable and indirect markers of oocyte competence are hyaluronan synthase 2 (HAS2), inhibin betaA (INHBA), epidermal growth factor receptor (EGFR), gremlin 1 (GREM1), betacellulin (BTC), CD44, tumor necrosis factor-induced protein 6 (TNFAIP6), and prostaglandin-endoperoxide synthase 2 (PTGS2). These biomarkers could be potential candidates to predict oocyte competence and to select higher-quality embryos for transfer. Additionally, these indirect predictors of oocyte competence and follicular health could improve our knowledge of gene expression patterns in the cumulus and yield insights into the molecular pathways controlling oocyte competence.
    Biology of Reproduction 05/2008; 79(2):209-22. · 4.03 Impact Factor

Publication Stats

160 Citations
17.78 Total Impact Points

Institutions

  • 2010
    • Université de Montréal
      Montréal, Quebec, Canada
  • 2008–2009
    • Laval University
      • • Département des Sciences Animales
      • • Centre de Recherche en Biologie de la Reproduction (CRBR)
      Québec, Quebec, Canada
    • Université du Québec
      • Département des Sciences Animales
      Québec, Quebec, Canada