Maintenance of Bovine Oocytes: MATERIALS AND METHODS(4)

MATERIALS AND METHODS(4)

Aspiration and Selection of Oocytes

After dissecting and washing of the ovaries, COCs were aspirated from follicles of 3-8 mm in diameter using a 10ml syringe fitted with an 18-gauge hypodermic needle. The aspirated follicular fluid was placed into sterile plastic universal containers in a warmed chamber (35°C) and allowed to settle for 10-15 min. asthma inhalers

The majority of the fluid was then removed by surface aspiration, and the remaining follicular material was diluted with an equal volume of dissection medium (Tissue Culture Medium 199 [TCM199] with Earle’s salts [Gibco, Grand Island, NY], 75.0 mg/L kana-mycin monosulfate [Sigma], 7.08 g/L Hepes [pH 7.8, os-molarity 279 mOsmol/kg H2O]) supplemented with 10% FCS. The diluted follicular fluid was transferred into an 85mm petri dish and examined for COCs under a dissecting microscope (X40 magnification). Oocytes were selected on morphological criteria; good-quality oocytes with homogenous evenly distributed cytoplasm and 3-4 layers of compact cumulus investment were selected for maturation.

This entry was posted in Bovine Oocytes and tagged Antral Follicle Culture, Bovine Oocytes, Meiotic Arrest.