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Procedure for rapid oocyte selection based on quantitative analysis of cumulus cell gene expression

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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To develop a procedure for the analysis of gene expression in cumulus cells during the interval between ovum pick up and insemination to select the best oocytes for fertilization.

Methods

Five RNA extraction methods, three reverse transcription procedures followed by Real-time quantitative PCR and one single-step mRNA quantification kit were tested to measure the expression of five genes in cumulus cells.

Results

Two RNA extraction kits gave the best combination of efficiency and purity. One reverse transcription procedure gave the best speed and efficiency. The single-step kit required more biological material than would be available from single cumulus oocyte complexes (COCs).

Conclusions

Our test identified a combination of RNA extraction and reverse transcription procedures that enables the level measurement of 5 selected cumulus cell transcripts within 4 h. Using this combination it was possible to obtain a reliable quantification of gene expression in 44 out of 46 individual COCs collected from seven patients.

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Acknowledgements

This work was supported by Regione Lombardia “Studi e progetti innovativi per la procreazione medicalmente assistita”.

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Authors and Affiliations

  1. Servizi Sforza srl, Medicina della Riproduzione, Milan, Italy

    Stefania Ferrari

  2. Department of Mother and Infant Sciences, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, University of Milan, Milan, Italy

    Debora Lattuada & Giorgio Bolis

  3. Infertility Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy

    Stefania Ferrari, Alessio Paffoni, Claudia Scarduelli & Guido Ragni

  4. UniSTEM, Laboratory of Biomedical Embriology, Università degli Studi di Milano, via Celoria 10, 20133, Milan, Italy

    Tiziana A. L. Brevini & Fulvio Gandolfi

Authors
  1. Stefania Ferrari
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  2. Debora Lattuada
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  3. Alessio Paffoni
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  4. Tiziana A. L. Brevini
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  5. Claudia Scarduelli
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  6. Giorgio Bolis
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  7. Guido Ragni
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  8. Fulvio Gandolfi
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Corresponding author

Correspondence to Stefania Ferrari.

Additional information

Capsule Development of a procedure for oocyte selection during the interval between ovum pick-up and insemination, based on gene expression levels in cumulus cells.

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Ferrari, S., Lattuada, D., Paffoni, A. et al. Procedure for rapid oocyte selection based on quantitative analysis of cumulus cell gene expression. J Assist Reprod Genet 27, 429–434 (2010). https://doi.org/10.1007/s10815-010-9428-1

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  • DOI: https://doi.org/10.1007/s10815-010-9428-1

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