In vitro maturation, fertilization and early cleavage rates of bovine fetal oocytes.
The in vitro developmental competence of oocytes harvested from 3 to 6 mm follicles from ovaries of 7.5 months to term fetuses and adult cows was compared. Cumulus oocyte complexes (COCs) were washed and placed in 200 microl droplets of maturation medium 199, supplemented with 10 microg/ml FSH, 10 microg/ml LH, 1.5 microg/ml estradiol, 75 microg/ml streptomycin, 100 IU/ml penicillin, 10 mM Hepes, and 10% fetal bovine serum (FBS) under oil and incubated for 24 h at 39 degrees C and 5% CO2. Matured oocytes were exposed to frozen-thawed TALP swim-up, heparin-capacitated sperm (20 h, 39 degrees C, 5% CO2). Presumptive zygotes were cultured in medium 199 containing 8 mg/ml BSA-V, 100 IU/ml penicillin G, 75 microg/ml streptomycin, and 10 mM Hepes (48 h, 39 degrees C, 5% CO2). Oocytes/embryos were fixed, stained with DAPI, and evaluated under fluorescent microscopy to assess maturation, fertilization, and subsequent embryonic development. There was a difference (P<0.05) between fetal and adult cow oocytes for in vitro maturation (IVM; 80.1% versus 92.0%), fertilization (69.3% versus 79.9%), and cleavage rates (36.7% versus 49.9%), respectively. Poor IVM, fertilization and embryonic development of fetal oocytes may be due to a higher incidence of blockage at germinal vesicle (GV) and metaphase-I (M-I) stage after IVM (12.0% versus 2.3% for fetal versus adult oocytes, respectively, P<0.05). Although the IVF results with fetal oocytes are poorer than with adult cow oocytes, they were still high enough to be considered for use in research and when death of the dam and/or fetus is pre-mature or sudden.
Chohan
Kazim R
KR
Division of Theriogenology, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108, USA. kazimchohan@hotmail.com
Hunter
Alan G
AG
D000818:Animals; D002417:Cattle / Q000196:embryology*; D002970:Cleavage Stage, Ovum; D003470:Culture Media; D046508:Culture Techniques; D005314:Embryonic and Fetal Development; D004958:Estradiol / Q000008:administration & dosage; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D005865:Gestational Age; D008297:Male; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000254:growth & development / Q000502:physiology*; D010053:Ovary / Q000166:cytology / Q000196:embryology*; D010406:Penicillins / Q000008:administration & dosage; D011247:Pregnancy; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000502:physiology; D013307:Streptomycin / Q000008:administration & dosage; D015053:Zygote / Q000502:physiology
D003470:Culture Media; D010406:Penicillins; D004958:Estradiol; D013307:Streptomycin