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a Departments of Obstetrics and Gynaecology and Physiology,
b The University of Western Ontario, London, Ontario, Canada N6A 5C1
c Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, Texas 77843-4466
The objectives were 1) to investigate the effects of oocyte maturation in serum-free and amino acid-supplemented defined media on oocyte transcript levels, blastocyst cell number, and apoptosis; 2) to investigate the influence of oocyte maturation culture atmosphere on blastocyst development, total cell number, and apoptosis; and 3) to examine the influence of epidermal growth factor (EGF) during oocyte maturation on blastocyst cell number and apoptosis. The results demonstrate that blastocysts derived from in vitro maturation, fertilization, and embryo culture protocols undergo apoptosis but that apoptotic levels are not greatly influenced by the oocyte maturation environment. Amino acid supplementation of oocyte maturation media was associated with enhanced developmental frequencies, increased blastocyst cell number, and elevated oocyte maternal mRNA levels. Oocyte maturation with supplemented synthetic oviduct fluid medium (cSOFMaa) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199 + newborn calf serum. Blastocyst development was reduced following oocyte maturation under a 5% CO2, 7% O2, 88% N2 culture atmosphere. EGF supplementation of oocyte maturation medium resulted in a concentration-dependent increase in blastocyst development but did not influence blastocyst total cell number or apoptosis. Our findings indicate that cSOFMaa medium is an effective base medium for bovine oocyte maturation.
1 This work was supported by the National Institute of Child Health and Human Development, National Cooperative Program on Non-Human In Vitro Fertilization and Embryo Development, USA. P.D. was supported by an Organon, Canada, Inc., Postdoctoral Fellowship in Reproductive Endocrinology; L.C.B. is the recipient of a NSERC Postgraduate Scholarship.
2 Correspondence. FAX: 519 661 3827;awatson{at}physiology.uwo.ca
3 Current address: Geron Bio-Med, Roslin Institute, Roslin, Midlothian, UK.
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