Category Archives: Bovine Oocytes - Part 2

Maintenance of Bovine Oocytes: MATERIALS AND METHODS(6)

Test of Oocyte Reversibility After 24-h culture, isolated COCs were washed twice in dissection medium and transferred into 500 ^l maturation medium (TCM199 with Earle’s salts [Gibco], 75 mg/L kanamycin, 4.75 g/L Hepes, 2.29 g/L NaHCO3 [pH 7.8, osmolarity 280 … Continue reading

Maintenance of Bovine Oocytes: MATERIALS AND METHODS(5)

Test of Oocyte Viability and Cell Cycle Stage After 24 h, oocytes from all culture procedures were recovered. A proportion of the oocytes were tested for viability by dye exclusion, using 0.5% trypan blue (Sigma; cell culture tested). Viable oocytes … Continue reading

Maintenance of Bovine Oocytes: 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 … Continue reading

Maintenance of Bovine Oocytes: MATERIALS AND METHODS(3)

Culture of Oocytes Attached to Part of the Follicle Wall Follicles were dissected from the ovaries as explained above. Selected follicles were cut according to De Loos et al.. Briefly, the COC was located using a stereomicro-scope. Directly opposite the … Continue reading

Maintenance of Bovine Oocytes: MATERIALS AND METHODS(2)

Fetal calf serum (FCS; 5%) was added prior to gassing. A gaseous atmosphere of 45% O2:50% N2:5% CO2 was used according to the method of Moor et al.. Selected follicles were transferred into 6-well culture dishes with or without netwell … Continue reading

Maintenance of Bovine Oocytes: MATERIALS AND METHODS(1)

Culture of Antral Follicles Ovaries were obtained from local abattoirs and maintained at 28-35°C during transport to the laboratory (1-2 h). Ovaries were dissected from the rest of the reproductive tract and transferred into a beaker containing 200-300 ml of … Continue reading

Maintenance of Bovine Oocytes in Meiotic Arrest and Subsequent Development In Vitro(3)

Other studies have attempted to establish physiological methods for the maintenance of meiotic arrest, including addition of follicular fluid to the culture medium and culture of oocytes on monolayers of granulosa or theca cells, within follicle hemisections, or attached to … Continue reading

Maintenance of Bovine Oocytes in Meiotic Arrest and Subsequent Development In Vitro(2)

In vivo, meiotic maturation of the oocytes occurs within the follicle. However, in vitro, when oocytes are removed from the follicle, meiotic maturation resumes spontaneously. The relationship between developmental competence and oocyte development is unclear. However, it is hypothesized that … Continue reading

Maintenance of Bovine Oocytes in Meiotic Arrest and Subsequent Development In Vitro(1)

The in vitro maturation of bovine oocytes coupled to fertilization and culture is a useful tool for both agricultural and research purposes (for review see ). Modifications to the culture conditions used for oocyte maturation, fertilization, and embryo development have … Continue reading

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