Culture of in vitro produced bovine zygotes in vitro vs in vivo: implications for early embryo development and quality.
The objectives of this study were to examine the effect of culture system on bovine blastocyst formation rates and quality. Presumptive IVM/IVF bovine zygotes were cultured either in vitro in synthetic oviduct fluid (SOF, 25 embryos/25 microL in 5% CO2, 5% O2, 90% N2 at 39 degrees C) or in vivo in the ewe oviduct (approximately 100 embryos per oviduct). The recovery rate after in vivo culture was 53% (813/1,530). The blastocyst rate on Day 7 was significantly higher for the in vitro system (28%, 362/1,278 vs 17%, 37/813; P< 0.0001). However, after culture in vitro for a further 24 h, there was no difference in Day 8 yields (36%, 457/1,278 vs 32%, 258/813, for in vitro and in vivo culture, respectively). There was no difference in blastocyst cell number between treatments (Day 7: 96 vs 103; Day 8: 78 vs 85 for in vitro and in vivo culture, respectively). Irrespective of culture system, Day 7 blastocysts had a significantly higher cell number than those appearing on Day 8. There was no difference in pregnancy rate at Day 35 after fresh transfer of a single Day 7 blastocyst (37.5%, 21/56 vs 45.3%/, 24/53 for in vitro and in vivo culture, respectively). After cryopreservation by freezing in 10% glycerol, VS3a vitrification or solid surface vitrification, the survival of in vitro cultured embryos was significantly lower than survival of embryos cultured in the ewe oviduct or those produced by superovulation of donors. In conclusion, these findings demonstrate that while bovine zygotes cultured in vitro are capable of rates of development similar to those of their in vivo cultured counterparts (in terms of Day 8 blastocyst yield, cell number and early pregnancy rate), there are significant differences in embryo cryosurvival. This suggests that current in vitro culture systems need to be improved to optimize embryo quality and pregnancy rates.
Enright
B P
BP
Department of Animal Science and Production, University College Dublin, Lyons Research Farm, Newcastle, Ireland.
Lonergan
P
P
Dinnyes
A
A
Fair
T
T
Ward
F A
FA
Yang
X
X
Boland
M P
MP
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology / Q000502:physiology*; D018929:Cell Culture Techniques; D003008:Cloprostenol / Q000008:administration & dosage; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004624:Embryo Transfer / Q000662:veterinary*; D005314:Embryonic and Fetal Development / Q000502:physiology; D005187:Fallopian Tubes / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D005990:Glycerol / Q000008:administration & dosage; D006062:Gonadotropins / Q000008:administration & dosage; D008297:Male; D011247:Pregnancy; D012756:Sheep; D015053:Zygote / Q000502:physiology*
D003451:Cryoprotective Agents; D006062:Gonadotropins; D003008:Cloprostenol; D005990:Glycerol