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Selection of Rattus norvegicus cumulus–oocyte complex for vitrification by brilliant cresyl blue

Published online by Cambridge University Press:  14 July 2023

Iaskara Oliveira
PPG – Ciências da Saúde − Universidade Federal de Ciências da Saúde de Porto Alegre –UFCSPA, Porto Alegre, RS, Brazil
Joana Fisch
PPG – Ciências da Saúde − Universidade Federal de Ciências da Saúde de Porto Alegre –UFCSPA, Porto Alegre, RS, Brazil
Juliana Gomes
Laboratório de Biotecnologia Animal Aplicada− Universidade Federal do Rio Grande do Sul−UFRGS, Porto Alegre, RS, Brazil
Rui Fernando Felix Lopes
Laboratório de Biotecnologia Animal Aplicada− Universidade Federal do Rio Grande do Sul−UFRGS, Porto Alegre, RS, Brazil
Alexandre Tavares Duarte de Oliveira*
PPG – Ciências da Saúde − Universidade Federal de Ciências da Saúde de Porto Alegre –UFCSPA, Porto Alegre, RS, Brazil Laboratório de Biotecnologia Animal Aplicada− Universidade Federal do Rio Grande do Sul−UFRGS, Porto Alegre, RS, Brazil
Corresponding author: Alexandre Tavares Duarte de Oliveira; Email:


The influence of the method of evaluating developmentally competent oocytes on their viability after cryopreservation still needs to be better understood. The objective of this study was to determine the cleavage and embryo developmental rates after parthenogenetic activation of cumulus–oocyte complexes (COCs) selected by different concentrations of brilliant cresyl blue (BCB) and cryopreservation. In the first experiment, COCs were separated into groups and incubated for 1 h in medium containing BCB (13 μM, 16 μM, or 20 μM). The control group was not exposed to BCB staining. In the second experiment, COCs were divided into four groups: 13 μM BCB(+), 13 μM BCB(−), fresh control (selected by morphologic observation and immediately in vitro matured) and vitrified control (selected by morphologic evaluation, vitrified, and in vitro matured). In the first experiment, the 13 μM BCB group displayed greater development rates at the morula stage (65.45%, 36/55) when compared with the other groups. In the second experiment, cleavage (47.05%, 72/153) and morula development (33.55%, 51/153) of the control group of fresh COCs were increased compared with the other groups. However, when comparing morula rates between vitrified COC control and BCB(+) groups, the BCB(+) group had better results (19.23%, 5/26 and 64.7%, 11/17, respectively). Our best result in rat COC selection by BCB staining was obtained using a concentration of 13 μM. This selection could be a valuable tool to improve vitrification outcomes, as observed by the BCB(+) group that demonstrated better results compared with the vitrified COC control.

Research Article
© The Author(s), 2023. Published by Cambridge University Press

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