The outcome of in-vitro-maturation and in-vitro-fertilization of feline oocytes depends on the selection of high quality oocytes, and is often restricted to morphological criteria. The aim of this study was to test whether the Brilliant cresyl blue (BCB) staining is suitable for pre-selection of feline oocytes before in-vitro-maturation. Cumulus-oocytes-complexes (COC) were released from domestic cat ovaries obtained after ovariectomy and were subjected to BCB staining. BCB+ stained oocytes were characterized by a violet/pale blue staining of the ooplasma, BCB- oocytes remained unstained. Transmission electron microscopy indicated for a slightly advanced stage of BCB- oocytes within the maturation process. After 24 h in-vitro-maturation, almost 75% of BCB+ and 21.5% of BCB- oocytes were able to reach metaphase II. Also, after in-vitro-fertilization, significantly more oocytes developed to morulae (19.2%) if oocytes were preselected for BCB staining, although 8% of unstained COC still reached advanced embryo stages. Prolonged storage of ovaries before COC retrieval for 16-20 h at 4 °C was accompanied by reduced number of BCB+ oocytes (96 of 210, 45.7%) in comparison to freshly isolated COC (151 of 225, 67.1%), and impaired cleavage rate (19.8%) and morula rate (9.4%) of BCB+ oocytes but the rate of embryos which developed to advanced stages remained unchanged (∼50%). To conclude, BCB staining is a very useful tool to preselect immature COC of feline species ensuring higher developmental rates.
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http://dx.doi.org/10.1016/j.theriogenology.2018.12.021 | DOI Listing |
Zygote
October 2024
Animal Reproduction Division, ICAR- Indian Veterinary Research Institute (ICAR-IVRI), Izatnagar243122, India.
The study was conducted on indigenous Tharparkar cow () to evaluate FSH stimulation on follicular attributes, oocyte recovery and morpho-molecular developmental competence parameters concerning oocyte quality. A total of 20 OPU sessions were performed, which included 10 sessions in each FSH stimulated at the dose of 130 µg divided into four sub-doses and non-stimulated. Findings on the size of follicles having ≥6 mm showed a significantly higher, however an opposite trend was observed in the case of smaller sized follicle (<6 mm) between stimulated and non-stimulated respectively.
View Article and Find Full Text PDFReprod Biol
September 2024
Department of Animal Reproduction, Anatomy and Genomics, University of Agriculture in Kraków, Al. Mickiewicza 24/28, 30-059 Kraków, Poland.
The aim of the study was to investigate the relationship between ooplasm morphology, lipid content, glucose-6-phosphate dehydrogenase activity (G6PDH) and maturation potential of domestic cat oocytes. Cumulus-oocyte complexes were classified according to ooplasm morphology: evenly dark (dCOC), heterogeneous/mosaic (hCOC), or light/transparent (lCOC), however only dCOCs are thought to be the best-quality, the remaining ones are usually rejected, therefore little is known about their intracellular properties. Lipid droplets (LDs) were visualized and quantified using Oil Red O.
View Article and Find Full Text PDFReprod Domest Anim
August 2024
Guangxi Key Laboratory of Buffalo Genetics, Reproduction and Breeding, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, China.
This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB-) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress-related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH-DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant-related genes SOD2 and GPX1.
View Article and Find Full Text PDFTheriogenology
September 2024
Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan. Electronic address:
Objectives of the present study were to investigate the characteristics including glucose-6-phosphate dehydrogenase activity, as determined by Brilliant Cresyl Blue (BCB) staining, of suboptimal porcine oocytes and to enhance the meiotic competence of those through pre-culture with cumulus cell masses (CCMs). Percentage of oocyte-cumulus complexes (OCCs) derived from small follicles (SF; <3 mm in diameter) containing the oocytes that were assessed as BCB-negative (BCB-) was significantly higher than those derived from medium follicles (MF; 3-6 mm in diameter). Degrees of dead cumulus cells were significantly higher in OCCs containing BCB- oocytes, regardless of the origin of OCCs (MF vs.
View Article and Find Full Text PDFNPJ Breast Cancer
April 2024
Laboratory for Translational Breast Cancer Research, Department of Oncology, KU Leuven, Leuven, Belgium.
Research on metastatic cancer has been hampered by limited sample availability. Here we present the breast cancer post-mortem tissue donation program UPTIDER and show how it enabled sampling of a median of 31 (range: 5-90) metastases and 5-8 liquids per patient from its first 20 patients. In a dedicated experiment, we show the mild impact of increasing time after death on RNA quality, transcriptional profiles and immunohistochemical staining in tumor tissue samples.
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