Effect of the in vitro culture system on the kinetics of blastocyst development and sex ratio of bovine embryos.
Bovine blastocysts were produced using 6 different systems: 5 commonly used in vitro culture systems (synthetic oviduct fluid medium - SOF- without fetal calf serum, SOF supplemented with 10% serum for the entire culture period, SOF supplemented with 10% serum from Day 4 of culture, M199 coculture with bovine oviduct epithelial cells, M199 coculture with granulosa cell monolayer) and 1 in vivo culture system involving collection of blastocysts from superovulated bovine donors at Day 7. Zygotes obtained from IVM/IVF were assigned randomly to 1 of the 5 systems tested and were cultured for 9 d (Day 0= day of insemination). Cleavage, development to the blastocyst stage and blastocyst sex ratio were assessed in all treatments. In addition, the effect of the IVC system on the kinetics of blastocyst development and sex ratio was assessed on Days 6, 7, 8, and 9. The presence of fetal calf serum in SOF not only resulted in faster development (19.1% of blastocysts in SOF supplemented with serum vs 7.1% in absence of serum at Day 6; P < 0.05) and increased blastocyst production (47.5% of blastocysts in SOF supplemented with serum vs 34.4% in absence of serum; P < 0.05) but it also enhanced overall male survival. The coculture systems produced fewer blastocysts than culture in SOF (27.6 to 28.3% in coculture vs 47.5% in SOF supplemented with serum; P < 0.05), but similar to SOF without fetal calf serum, they had no effect on blastocyst sex ratio.
Gutiérrez-Adán
A
A
Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Madrid, Spain.
Lonergan
P
P
Rizos
D
D
Ward
F A
FA
Boland
M P
MP
Pintado
B
B
de la Fuente
J
J
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002417:Cattle / Q000502:physiology*; D018920:Coculture Techniques; D003470:Culture Media; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D005312:Fetal Blood; D008297:Male; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011247:Pregnancy; D011897:Random Allocation; D012732:Sex Determination Analysis / Q000662:veterinary; D012744:Sex Ratio
D003470:Culture Media