title: Procedure for rapid oocyte selection based on quantitative analysis of cumulus cell gene expression.
abstract: OBJECTIVE
  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.
authors.affiliation: Servizi Sforza srl, Medicina della Riproduzione, Milan, Italy. centrosterilita@policlinico.mi.it
authors.forename: Stefania
authors.initials: S
authors.lastname: Ferrari
authors.forename: Debora
authors.initials: D
authors.lastname: Lattuada
authors.forename: Alessio
authors.initials: A
authors.lastname: Paffoni
authors.forename: Tiziana A L
authors.initials: TA
authors.lastname: Brevini
authors.forename: Claudia
authors.initials: C
authors.lastname: Scarduelli
authors.forename: Giorgio
authors.initials: G
authors.lastname: Bolis
authors.forename: Guido
authors.initials: G
authors.lastname: Ragni
authors.forename: Fulvio
authors.initials: F
authors.lastname: Gandolfi
mesh_terms: D054885:Cumulus Cells / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression; D006801:Humans; D054315:Oocyte Retrieval; D009865:Oocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D012333:RNA, Messenger
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