This study assessed the impact of various cryoprotectant (CPA) exposures on nuclear and cytoplasmic maturation in the immature cat oocyte as a prerequisite to formulating a successful cryopreservation protocol. In experiment 1, immature oocytes were exposed to 0, 0.75, 1.5, or 3.0 M of 1,2-propanediol (PrOH) or 1,2-ethanediol (EG) at room temperature (25°C) or 0°C for 30 min. After CPA removal and in vitro maturation, percentage of oocytes reaching metaphase II (MII) was reduced after exposure to 3.0 M PrOH at 0°C or 3.0 M EG at both temperatures. All CPA exposures increased MII spindle abnormalities compared to control, except 1.5 M PrOH at 25°C. In experiments 2 and 3, immature oocytes were exposed to CPA conditions yielding optimal nuclear maturation that either had caused spindle damage (0.75 M PrOH, 1.5 M EG, and 3.0 M PrOH at 25°C) or not (1.5 M PrOH at 25°C). After maturation and insemination in vitro, oocytes were cultured for 7 days to assess treatment influence on developmental competence. CPA exposure did not affect fertilization, but the high incidence of MII spindle abnormalities resulted in a low percentage of cleaved embryos. Blastocyst formation and quality were influenced by both CPA types (EG was more detrimental than PrOH) and concentration (3.0 M was more detrimental than 1.5 M). Overall, cat oocytes appear to be highly sensitive to CPA except after exposure to 1.5 M PrOH at 25°C, a treatment that still allowed ∼60% of the oocytes to reach MII and ∼20% to form blastocysts.
How to translate text using browser tools
1 August 2004
Effect of 1,2-Propanediol Versus 1,2-Ethanediol on Subsequent Oocyte Maturation, Spindle Integrity, Fertilization, and Embryo Development In Vitro in the Domestic Cat
Pierre Comizzoli,
David E. Wildt,
Budhan S. Pukazhenthi
ACCESS THE FULL ARTICLE
assisted reproductive technology
early development
in vitro fertilization
meiosis
toxicology