Skip to main content Accessibility help

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

  • Diego Duarte Alcoba (a1), Bianca Letícia da Rosa Braga (a1), Nathallie Louise Sandi-Monroy (a1), Letícia Auler Proença (a1), Rui Fernando Felix Lopes (a2) and Alexandre Tavares Duarte de Oliveira (a3)...


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.


Corresponding author

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:


Hide All
Adona, P.R., Pires, P.R., Quetglas, M.D., Schwarz, K.R. & Leal, C.L. (2008). Prematuration of bovine oocytes with butyrolactone I: effects on meiosis progression, cytoskeleton, organelle distribution and embryo development. Anim. Reprod. Sci. 108, 4965.
Alm, H., Torner, H., Lohrke, B., Viergutz, T., Ghoneim, I.M. & Kanitz, W. (2005). Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brillant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. Theriogenology 63, 2194–205.
Bhojwani, S., Alm, H., Torner, H., Kanitz, W. & Poehland, R. (2007). Selection of developmentally competent oocytes through brilliant cresyl blue stain enhances blastocyst development rate after bovine nuclear transfer. Theriogenology 67, 341–5.
Dekel, N. (1995). Molecular control of meiosis. Trends Endocrinol. Metab. 6, 165–9.
Dekel, N. & Beers, W.H. (1978). Rat oocyte maturation in vitro: relief of cyclic AMP inhibition by gonadotropins. Proc. Natl. Acad. Sci. USA 75, 4369–73.
Goovaerts, I.G., Leroy, J.L., Jorssen, E.P. & Bols, P.E. (2010). Noninvasive bovine oocyte quality assessment: possibilities of a single oocyte culture. Theriogenology 74, 1509–20.
Grøndahl, C. (2008). Oocyte maturation. Basic and clinical aspects of in vitro maturation (IVM) with special emphasis of the role of FF-MAS. Dan. Med. Bull. 55, 116.
Hyttel, P., Fair, T., Callesen, H. & Greve, T. (1997). Oocyte growth, capacitation and final maturation in cattle. Theriogenology 47, 2332.
Jiang, J.Y., Xiong, H., Cao, M., Xia, X., Sirard, M.A. & Tsang, B.K. (2010). Mural granulosa cell gene expression associated with oocyte developmental competence (abstract). J. Ovarian Res. 3, 6.
Mangia, F. & Epstein, C.J. (1975). Biochemical studies of growing mouse oocytes – preparation of oocytes and analysis of glucose-6-phosphate-dehydrogenase and lactate-dehydrogenase activities. Dev. Biol. 45, 211–20.
Manjunatha, B.M., Gupta, P.S., Devaraj, M., Ravindra, J.P. & Nandi, S. (2007). Selection of developmentally competent buffalo oocytes by brilliant cresyl blue staining before IVM. Theriogenology 68, 12991304.
Marcondes, F.K., Bianchi, F.J. & Tanno, A.P. (2002). Determination of the estrous cycle phases of rats: some helpful considerations. Braz. J. Biol. 62, 609–14.
Mota, G.B., Batista, R.I., Serapião, R.V., Boité, M.C., Viana, J.H., Torres, C.A. & de Almeida Camargo, L.S. (2010). Developmental competence and expression of the MATER and ZAR1 genes in immature bovine oocytes selected by brilliant cresyl blue. Zygote 18, 209–16.
Opiela, J., Katska-Ksiazkiewicz, L., Lipiński, D., Słomski, R., Bzowska, M. & Ryńska, B. (2008). Interactions among activity of glucose-6-phosphate dehydrogenase in immature oocytes, expression of apoptosis-related genes Bcl-2 and Bax, and developmental competence following IVP in cattle. Theriogenology 69, 546–55.
Opiela, J., Lipiński, D., Słomski, R. & Katska-Ksiazkiewicz, L. (2010). Transcript expression of mitochondria related genes is correlated with bovine oocyte selection by BCB test. Anim. Reprod. Sci. 118, 188–93.
Quinn, P., Barros, C. & Whittingham, D.G. (1982). Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J. Reprod. Fertil. 66, 161–8.
Rodrigues, B.A., Rodriguez, P., Silva, A.E., Cavalcante, L.F., Feltrin, C. & Rodrigues, J.L. (2009). Preliminary study in immature canine oocytes stained with brilliant cresyl blue and obtained from bitches with low and high progesterone serum profiles. Reprod. Domest. Anim. 44, 255–8.
Rodriguez-González, E., Lóopez-Béjar, M., Velilla, E. & Paramio, M.T. (2002). Selection of prepubertal goat oocytes using the brilliant cresyl blue test. Theriogenology 57, 13971409.
Torner, H., Ghanem, N., Ambros, C., Holker, M., Tomek, W., Phatsara, C., Alm, H., Sirard, M.A., Kanitz, W., Schellander, K. & Tesfaye, D. (2008). Molecular and subcellular characterisation of oocytes screened for their developmental competence based on glucose-6-phosphate dehydrogenase activity. Reproduction 135, 197212.
Tsutsumi, O., Satoh, K., Taketani, Y. & Kato, T. (1992). Determination of enzyme activities of energy metabolism in the maturing rat oocyte. Mol. Reprod. Dev. 33, 333–7.
Whittingham, D.G. (1971). Culture of mouse ova. J. Reprod. Fertil. Suppl. 14, 721.
Wongsrikeao, P., Otoi, T., Yamasaki, H., Agung, B., Taniguchi, M., Naoi, H., Shimu, R. & Nagai, T. (2006). Effects of single and double exposure to brilliant cresyl blue on the selection of porcine oocytes for in vitro production of embryos. Theriogenology 66, 366–72.
Wu, Y.G., Liu, Y., Zhou, P., Lan, G.C., Han, D., Miao, D.Q. & Tan, J.H. (2007). Selection of oocytes for in vitro maturation by brilliant cresyl blue staining: a study using the mouse model. Cell Res. 17, 722–31.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed