Improved in vitro embryo development using in vivo matured oocytes from heifers superovulated with a controlled preovulatory LH surge.
In bovine in vitro embryo production, the IVM step is rather successful with 80% of the oocytes reaching the MII stage. However, the extent to which the process limits the yield of viable embryos is still largely unknown. Therefore, we compared embryonic developmental capacity during IVC of IVF oocytes which had been matured in vitro with those matured in vivo. In vitro maturation was carried out for 22 h using oocytes (n = 417) obtained from 2- to 8-mm follicles of ovaries collected from a slaughterhouse in M199 with 10% fetal calf serum (FCS), 0.01 IU/mL LH, and 0.01 IU/mL FSH. In vivo matured oocytes (n = 219) were aspirated from preovulatory follicles in eCG/PG/anti-eCG-superovulated heifers 22 h after a fixed time GnRH-induced LH surge; endogenous release of the LH surge was suppressed by a Norgestomet ear implant. This system allowed for the synchronization of the in vitro and in vivo maturation processes and thus for simultaneous IVF of both groups of oocytes. The in vitro developmental potential of in vivo matured oocytes was twice as high (P < 0.01) as that of in vitro matured oocytes, with blastocyst formation and hatching rates 11 d after IVC of 49.3 +/- 6.1 (SEM; n = 10 heifers) vs 26.4 +/- 1.0% (n = 2 replicates), and 39.1 +/- 5.1% vs 20.6 +/- 1.4%, respectively. It is concluded that IVM is a major factor limiting in the in vitro production of viable embryos, although factors such as the lack of normal preovulatory development of IVM oocytes contributed to the observed differences.
van de Leemput
E E
EE
Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, The Netherlands.
Vos
P L
PL
Zeinstra
E C
EC
Bevers
M M
MM
van der Weijden
G C
GC
Dieleman
S J
SJ
D000003:Abattoirs; D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D004343:Drug Implants; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000378:metabolism* / Q000494:pharmacology; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects; D006080:Ovarian Follicle / Q000166:cytology; D011282:Pregnenediones / Q000494:pharmacology; D013480:Superovulation / Q000187:drug effects
D006063:Chorionic Gonadotropin; D004343:Drug Implants; D011282:Pregnenediones; C100284:norgestomet; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone