Selection of ovine oocytes by brilliant cresyl blue staining.

Journal of biomedicine & biotechnology (2012-06-08)
Liqin Wang, Jiapeng Lin, Juncheng Huang, Jing Wang, Yuncheng Zhao, Tong Chen
ABSTRACT

Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB-) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Mineral oil, suitable for preparation of Nujol mulls for infrared spectroscopy, light oil
Sigma-Aldrich
Brilliant Cresyl Blue ALD, certified by the Biological Stain Commission