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Selection of Rattus norvegicus oocytes for in vitro maturation by brilliant cresyl blue staining

Published online by Cambridge University Press:  27 July 2011

Diego Duarte Alcoba ,
Bianca Letícia da Rosa Braga ,
Nathallie Louise Sandi-Monroy ,
Letícia Auler Proença ,
Rui Fernando Felix Lopes  and
Alexandre Tavares Duarte de Oliveira
Diego Duarte Alcoba
Affiliation:
Laboratório de Biologia Celular–Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA)–Porto Alegre, Brazil.
Bianca Letícia da Rosa Braga
Affiliation:
Laboratório de Biologia Celular–Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA)–Porto Alegre, Brazil.
Nathallie Louise Sandi-Monroy
Affiliation:
Laboratório de Biologia Celular–Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA)–Porto Alegre, Brazil.
Letícia Auler Proença
Affiliation:
Laboratório de Biologia Celular–Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA)–Porto Alegre, Brazil.
Rui Fernando Felix Lopes
Affiliation:
Laboratório de Biotecnologia Animal Aplicada–Universidade Federal de Rio Grande do Sul (UFRGS)–Porto Alegre, Brazil.
Alexandre Tavares Duarte de Oliveira*
Affiliation:
Universidade Federal de Ciências da Saúde de Porto Alegre – UFCSPA, Rua Sarmento Leite, 245 – sala 06, Porto Alegre – Rio Grande do Sul, 90050-170, Brazil.
*
All correspondence to: Alexandre Tavares Duarte de Oliveira. Universidade Federal de Ciências da Saúde de Porto Alegre – UFCSPA, Rua Sarmento Leite, 245 – sala 06, Porto Alegre – Rio Grande do Sul, 90050-170, Brazil. Tel: +55 51 33 03 8823. Fax: +55 51 33 03 8810. e-mail: atdo@ufcspa.edu.br
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Summary

The objective of this work was to evaluate the rate of meiosis resumption and nuclear maturation of rat (Rattus norvegicus) oocytes selected for in vitro maturation (IVM) after staining of cumulus–oocyte complexes (COCs) with blue cresyl brilliant (BCB) using different protocols: exposure for 30, 60 or 90 min at 26 μM BCB (Experiment 1), and exposure for 60 min at 13, 20 or 26 μM BCB (Experiment 2). In Experiment 1, the selection of oocytes exposed to BCB for 60 min was found to be the most suitable, as meiosis resumption rates in the BCB+ group (n = 35/61; 57.37%) were the closest to the observed in the control (not exposed) group (n = 70/90; 77.77%) and statistically higher than the values observed for the BCB− group (n = 3/41; 7.32%). Additionally, the more effective evaluation of diagnostic tests (sensitivity and negative predictive value 100%) was observed in COCs exposed for 60 min. In Experiment 2, the 13 μM BCB+ group presented rates of meiosis resumption (n = 57/72; 72.22%) similar to the control group (n = 87/105; 82.86%) and higher than other concentration groups. However, this results of the analysis between BCB− oocytes was also higher in the 13 μM BCB group (n = 28/91; 30.78%) when compared with BCB− COCs exposed to 20 μM (n = 3/62; 4.84%) or 26 μM (n = 3/61; 4.92%) BCB. The nuclear maturation rate in the 13 μM BCB group was similar between BCB+ or BCB− oocytes. The 20 μM BCB group had a lower rate of nuclear maturation of BCB− oocytes than other groups. Thus, our best results in the selection of Rattus norvegicus oocytes by staining with BCB were obtained using the concentration of 13 μM and 20 μM, and an incubation period of 60 min.

Keywords

Brilliant cresyl blueIn vitro maturationOocytesRattus norvegicus
Type
Research Article
Information
Zygote , Volume 21 , Issue 3 , August 2013 , pp. 238 - 245
Copyright
Copyright © Cambridge University Press 2011 

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