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

Stefania Ferrari, Debora Lattuada, Alessio Paffoni, Tiziana A L Brevini, Claudia Scarduelli, Giorgio Bolis, Guido Ragni, Fulvio Gandolfi

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)429-434
Number of pages6
JournalJournal of Assisted Reproduction and Genetics
Volume27
Issue number7
DOIs
Publication statusPublished - Jul 2010

Keywords

  • Marker genes
  • Oocyte selection
  • Prospective study
  • Real-time PCR

ASJC Scopus subject areas

  • Obstetrics and Gynaecology
  • Reproductive Medicine
  • Developmental Biology
  • Genetics
  • Genetics(clinical)

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