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C57BL/6J mouse superovulation: schedule and age optimization to increase oocyte yield and reduce animal use

Published online by Cambridge University Press:  15 January 2021

Sofia Lamas Open the ORCID record for Sofia Lamas [Opens in a new window] ,
Júlio Carvalheira ,
Fátima Gartner  and
Irina Amorim
Sofia Lamas*
Affiliation:
i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal Instituto de Biologia Molecular e Celular – IBMC Rua Alfredo Allen 208, 4200-135 Porto, Portugal
Júlio Carvalheira
Affiliation:
Institute of Biomedical Science Abel Salazar, University of Porto, Portugal Research Center in Biodiversity and Genetic Resources (CIBIO-InBio), University of Porto, Vairão, Portugal
Fátima Gartner
Affiliation:
Glycobiology in Cancer, IPATIMUP/ICBAS/i3S, Porto, Portugal Institute of Biomedical Science Abel Salazar, University of Porto, Portugal
Irina Amorim
Affiliation:
Glycobiology in Cancer, IPATIMUP/ICBAS/i3S, Porto, Portugal Institute of Biomedical Science Abel Salazar, University of Porto, Portugal
*
Author for correspondence: Sofia Lamas. i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal. Tel: +351 226074979 E-mail: sofia.lamas@ibmc.up.pt
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Abstract

Superovulation protocols have been described for different mouse strains, however the numbers of animals used are still high and still little information is known about hormone administration schedules and estrous cycle phases. In this study, we aimed to optimize a superovulation protocol by injecting 5 IU of pregnant mare serum gonadotropin followed by 5 IU of hCG 48 h later, using three different schedules related to the beginning of the dark cycle (3, 5 and 7 pm) in a light cycle of 7 am to 7 pm, with light on at 7 am. C57BL/6J mice at 3, 4 and 5 weeks of age were used and the estrous cycle phase for times of PMSG and hCG injections was also analyzed. Total oocyte number was counted in the morning after hCG injection. Hormones given at 3 weeks of age at 3 pm (59 ± 15 oocytes) and 7 pm (61 ± 10 oocytes) produced a significantly higher oocyte number compared with oocytes numbers collected from females at the same age at 5 pm (P = 0.0004 and <0.0001 respectively). Females at 4 and 5 weeks of age produced higher numbers of oocytes when superovulated at 7 pm. No statistical differences between females at different phases of the estrous cycle were found. These results showed that in C57BL/6J mice, hormones should be given at 3 or 7 pm for females at 3 weeks of age, however older females should be superovulated closer to the beginning of the dark cycle to reduce female mouse use and increase the numbers of oocytes produced per female.

Keywords

AgeC57BL/6JEstrous cycleScheduleSuperovulation
Type
Research Article
Information
Zygote , Volume 29 , Issue 3 , June 2021 , pp. 199 - 203
Copyright
© The Author(s), 2021. Published by Cambridge University Press

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