Impact of bovine oocyte maturation media on oocyte transcript levels, blastocyst development, cell number, and apoptosis.
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% CO(2), 7% O(2), 88% N(2) 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.
Watson
A J
AJ
Departments of Obstetrics and Gynaecology and Physiology, The University of Western Ontario, London, Ontario, Canada N6A 5C1. awatson@physiology.uwo.ca
De Sousa
P
P
Caveney
A
A
Barcroft
L C
LC
Natale
D
D
Urquhart
J
J
Westhusin
M E
ME
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002417:Cattle; D002452:Cell Count; D004396:Coloring Agents; D016895:Culture Media, Serum-Free; D004247:DNA / Q000737:chemistry; D005314:Embryonic and Fetal Development / Q000502:physiology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro; D005819:Genetic Markers; D007091:Image Processing, Computer-Assisted; D020287:In Situ Nick-End Labeling; D066298:In Vitro Techniques; D018715:Microscopy, Video; D009865:Oocytes / Q000254:growth & development* / Q000378:metabolism*; D011419:Propidium; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000596:Amino Acids; D004396:Coloring Agents; D016895:Culture Media, Serum-Free; D005819:Genetic Markers; D012333:RNA, Messenger; D011419:Propidium; D004815:Epidermal Growth Factor; D004247:DNA