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.
Procedure for rapid oocyte selection based on quantitative analysis of cumulus cell gene expression / S. Ferrari, D. Lattuada, A. Paffoni, T. Brevini, C. Scarduelli, G. Bolis, G. Ragni, F. Gandolfi. - In: JOURNAL OF ASSISTED REPRODUCTION AND GENETICS. - ISSN 1058-0468. - 27:7(2010 Jul), pp. 429-434.
Titolo: | Procedure for rapid oocyte selection based on quantitative analysis of cumulus cell gene expression |
Autori: | |
Settore Scientifico Disciplinare: | Settore VET/01 - Anatomia degli Animali Domestici |
Data di pubblicazione: | lug-2010 |
Rivista: | |
Tipologia: | Article (author) |
Digital Object Identifier (DOI): | http://dx.doi.org/10.1007/s10815-010-9428-1 |
Appare nelle tipologie: | 01 - Articolo su periodico |
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