Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test

Pujol, Marc; López‐Béjar, Manel; Paramio, M. T.

doi:10.1016/s0093-691x(03)00250-4

Cited by 112 publications

(112 citation statements)

References 33 publications

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“…In this study, we have observed significantly higher 2PN zygote and blastocyst production in BCBC than BCBK oocytes. This increase in 2PN zygote and blastocyst development has been shown in BCBC oocytes of buffalo (Manjunatha et al 2007), cattle (Pujol et al 2004, Alm et al 2005, Bhojwani et al 2007, Torner et al 2008, goats (Rodriguez-Gonzalez et al 2002), mice (Wu et al 2007) and pigs (Egerszegi et al 2010).…”

Section: Discussionsupporting

confidence: 58%

“…In cows (Pujol et al 2004, Alm et al 2005, goats (Rodriguez-Gonzalez et al 2002) and mice (Wu et al 2007) the concentration of BCB stain with the best results was 26 mM BCB for 90 min, while in pigs 13 mM BCB was sufficient to increase sperm penetration of BCBC oocytes (Roca et al 1998, Egerszegi et al 2010. In our work, we decided to work with 13 mM BCB for 60 min, as it was the most specific concentration, increasing blastocyst rate from 9% (BCBK) to 21% (BCBC).…”

Section: Discussionmentioning

confidence: 95%

“…However, growing oocytes are expected to have a high level of G6PDH activity and be able to reduce the blue compound, resulting in a colourless oocyte cytoplasm (BCBK). In our previous studies in prepubertal goats (Rodriguez-Gonzalez et al 2002) and cows (Pujol et al 2004), we have shown the usefulness of the BCB stain to select the larger and more competent oocytes for in vitro blastocyst production.…”

Section: Introductionmentioning

confidence: 98%

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Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep

Catalá¹,

Izquierdo²,

Uzbekova³

et al. 2011

Reproduction

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The aim of this study was to test the Brilliant Cresyl Blue (BCB) stain to select prepubertal sheep oocytes for in vitro blastocyst production. Oocyte diameter, mitochondrial activity, maturation-promoting factor (MPF) activity and mRNA relative expression (RE) of genes related to metabolism (ATPase Na C /K C transporting a 1 (ATP1A1) and cytochrome c oxidase subunit 1 (COX1)) and constitutive function of the cell (cytoplasmic polyadenylation-element-binding protein (CPEB) and S100A10) were assessed. Immature oocytes were exposed to different BCB concentrations (13, 26, 39 and 52 mM) and classified according to their cytoplasm colouration as grown BCBC (blue cytoplasm) and growing BCBK (colourless cytoplasm). Staining oocytes with 13 mM BCB during 60 min allows selection of (BCBC) the largest (123.66 mm) and most competent oocytes to develop to the blastocyst stage (21%) with a higher number of cells (69.71G6.19S.E.M.) compared with non-stained BCBK oocytes (106.82 mm, 9% and 45.91G3.35 S.E.M. respectively). Mitochondrial activity, assessed by MitoTracker Orange CMTMRos probe, was significantly higher in BCBC than in BCBK oocytes after in vitro maturation (3369 and 1565 AU respectively). MPF activity was assessed by CDC2 kinase activity assay showing significantly higher activity at metaphase II stage in BCBC than in BCBK oocytes (1.479G0.09 and 1.184G0.05 optical density respectively). The genes analysed in this work, ATP1A1, COX1, CPEB and S100A10, did not show significant effect in mRNA RE between BCB selected oocytes. In conclusion, BCB stains larger and more competent oocytes to develop to the blastocyst stage with more active mitochondria and MPF activity and higher blastocyst cell number.

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 98%

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Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep

Catalá¹,

Izquierdo²,

Uzbekova³

et al. 2011

Reproduction

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“…Previous studies in our laboratory [4] have shown that prepubertal goat oocytes stained with BCB show significantly higher rates of nuclear maturation and normal fertilisation (2PN oocytes) than unstained oocytes (BCB-), although the percentage of blastocysts is still low (4%). Pujol et al [3] in calf oocytes have also shown increases in the percentages of the blastocyst yield of BCB+ compared to BCB-oocytes (12.3 vs. 1.6% respectively).…”

Section: Discussionmentioning

confidence: 87%

“…Thus, oocytes that have finished their growth phase show a decreased G6PD activity and exhibit a cytoplasm with a blue coloration because they do not reduce BCB to a colourless compound. The BCB test has been used successfully to select oocytes for IVM-IVF in pigs [1,2] and cows [3]. In our earlier study with prepubertal goat oocytes matured without cysteamine [4], we showed that oocytes exposed and stained with BCB are larger (136.6 vs. 125.5 µm in diameter) and more able to develop to morula and blastocysts than unstained oocytes (12.0 vs. 3.6%).…”

Section: Introductionmentioning

confidence: 85%

Developmental competence of prepubertal goat oocytes selected with brilliant cresyl blue and matured with cysteamine supplementation

et al. 2003

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-The aim of this study was to assess the effect of oocyte selection using the brilliant cresyl blue (BCB) test plus the addition of cysteamine to the in vitro maturation (IVM) medium to improve the in vitro embryo development of prepubertal goat oocytes. The oocytes were exposed to 26 µM BCB and classified according to their cytoplasm coloration: BCB+ (oocytes with blue cytoplasm) and BCB-(unstained oocytes). The oocytes were matured in a conventional IVM medium supplemented with cysteamine 100 µM. The control group consisted of oocytes not exposed to BCB and matured without cysteamine. The IVM-oocytes were inseminated and cultured in synthetic oviductal fluid (SOF) for 7 days. The normal fertilisation rate (oocytes showing 2 pronuclei and 1 sperm tail) of BCB+ oocytes (40%) was higher than those of BCB-(21%) and control oocytes (22%). The percentage of morulae plus blastocysts was higher (P < 0.05) in the BCB+ group than in the BCB-group (23.8 vs. 5.1%, respectively). In conclusion, the integration of the BCB test and the addition of cysteamine in the protocol of in vitro embryo production from prepubertal goat oocytes has improved the developmental rates of embryo development. embryo / IVF / IVM / thiol

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Induced cumulus expansion of poor quality buffalo cumulus oocyte complexes by Interleukin‐1beta improves their developmental ability

Chaubey

Kumar

et al. 2018

J of Cellular Biochemistry

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The present study was conceived with the aim of modulating the cumulus expansion characteristics of poor quality (BCB-) buffalo oocyte complexes (COCs) in order to improve their fertilization outcomes. BCB- COCs were subjected to in vitro maturation (IVM) in presence of Interleukin-1 beta (IL-1β) along with BCB- (control) and good quality (BCB+) COCs. Results were assessed morphologically, by scanning electron microscopy (SEM) and by expression analysis of cumulus expansion related genes. Also, numbers of zona pellucida bound spermatozoa were counted and development rates of oocytes were monitored under different groups. Expression of versican isoforms and ADAMTS-1 was observed to be significantly different between cumulus cells of BCB+ and BCB- COCs. Upon IL-1β supplementation, ADAMTS-1 expression increased in BCB- COCs along with corresponding cumulus expansion rates. SEM analysis also revealed improved cumulus expansion in IL-1β supplemented BCB- COCs. HAS2 and TNFAIP-6 were significantly up-regulated after IL-1β supplementation while PTGS2 expression remained unaffected. Significantly more numbers of sperms crossed the cumulus barrier, especially in 100 ng/mL IL-1β supplemented COCs. Besides, cleavage and blastocyst development rates were also improved upon IL-1β addition. We concluded that IL-1β supplementation in IVM medium can improve cumulus expansion and development ability of poor quality buffalo oocytes.

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Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test

Cited by 112 publications

References 33 publications

Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep

Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep

Developmental competence of prepubertal goat oocytes selected with brilliant cresyl blue and matured with cysteamine supplementation

Induced cumulus expansion of poor quality buffalo cumulus oocyte complexes by Interleukin‐1beta improves their developmental ability

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