Selection of Immature Bovine Oocytes Using Brilliant Cresyl Blue Enhances Nuclear Maturity after Vitrification

Hajarian, Hadi; Wahid, H.; Rosnina, Y.; Daliri, Morteza; Dashtizad, Mojtaba; Karamishabankareh, Hamed; Faizah, A.; Iswadi, M. I.; Mazni, O. Abas

doi:10.3923/javaa.2010.2710.2713

Cited by 5 publications

(2 citation statements)

References 18 publications

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“…Our study demonstrated that oocytes that show BCB staining at the time of isolation resulted in higher nuclear maturation (Fig. 3), as has been reported by other groups 15,16 . We also showed that aspiration of antral follicles resulted in a higher proportion of BCB + oocytes in comparison to slicing (Fig.…”

Section: Discussionsupporting

confidence: 89%

“…During their growth phase, oocytes have high G6PDH activity and this enzyme degrades the BCB compound, resulting in a reduction in blue colour 15 . On the other hand, oocytes nearing the end of their growth phase have reduced G6PDH activity, and therefore have more intense blue cytoplasmic staining 14–16 . BCB staining can therefore be used to determine maturation potential of COCs isolated by different selection methods.…”

Section: Introductionmentioning

confidence: 99%

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Amino acid supplementation of a simple inorganic salt solution supports efficient in vitro maturation (IVM) of bovine oocytes

Bahrami

Morris

Day

2019

Sci Rep

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Defining oocyte in vitro maturation (IVM) conditions allows for improved reproducibility and efficiency of bovine embryo production. IVM conditions for bovine oocytes have been extensively studied, but beneficial effects of individual supplements remain controversial. This study compared methods of cumulus oocyte complex (COC) isolation, and culture medium requirements, for IVM in order to define optimal conditions. Antral follicles in ovaries were sliced or aspirated to isolate COCs. Brilliant cresyl blue staining of COCs was used to determine the most effective collection technique and the effect of hormones and groups of amino acids in the culture medium was investigated. Our results showed COCs isolated through aspiration had greater meiotic competency to reach MII. Oocyte maturation was achieved with the addition of 1 µg/mL FSH, while estrogen and human chorionic gonadotrophin did not increase the number of MII oocytes. We also provide novel data, that supplementation of a simple inorganic salt solution with L-proline, L-glutamine and essential amino acids in combination, but not individually, resulted in nuclear maturation comparable to TCM199, a more complex medium containing all 20 common amino acids, vitamins, inorganic salts and FBS. Replacement of FBS with BSA in this simplified medium creates a defined medium which provides conditions for IVM that enable reproducible maturation rates.

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Section: Discussionsupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Amino acid supplementation of a simple inorganic salt solution supports efficient in vitro maturation (IVM) of bovine oocytes

Bahrami

Morris

Day

2019

Sci Rep

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Culture conditions for in vitro maturation of oocytes – A review

Bahrami

Cottee²

2022

Reproduction and Breeding

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

Oliveira

Fisch

Gomes

et al. 2023

Zygote

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Summary 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.

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Selection of Immature Bovine Oocytes Using Brilliant Cresyl Blue Enhances Nuclear Maturity after Vitrification

Cited by 5 publications

References 18 publications

Amino acid supplementation of a simple inorganic salt solution supports efficient in vitro maturation (IVM) of bovine oocytes

Amino acid supplementation of a simple inorganic salt solution supports efficient in vitro maturation (IVM) of bovine oocytes

Culture conditions for in vitro maturation of oocytes – A review

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

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