title: Post-thaw culture in presence of insulin-like growth factor I improves the quality of cattle cryopreserved embryos.
abstract: The goal of this study was to examine the effect of insulin-like growth factor I (IGF-I; added during post-thaw culture (48 h)) on the preimplantation viability and quality of cryopreserved bovine in vivo recovered embryos. The morula stage embryos, non-surgically recovered from superovulated dairy cows of Czech Fleckvieh cattle breed, had previously been cryopreserved by a slow freezing technique and stored in liquid nitrogen since 1989-1990. Following thawing, the embryos were cultured for 48 h either alone (no IGF-I) or in the presence of IGF-I (10 or 100 ng/ml); non-cultured embryos served as a control. Thereafter, the embryos were analyzed for cleavage to the blastocyst stage, apoptosis (TUNEL), embryo cell number and quality of actin cytoskeleton. Following post-thaw culture 41% of embryos developed to advanced blastocysts. IGF-I increased this per cent and, at a higher dose, essentially reduced the per cent of degenerated embryos. In cultured embryos, IGF-I at both doses elevated the cell number compared with non-cultured embryos. However, in comparison with embryos cultured without IGF-I, only the higher IGF-I dose resulted in elevating the embryo cell number. The TUNEL index was significantly lowered by IGF-I treatment. Thawed embryos were mostly of the grade III actin type and fewer (12%) had grade II actin, whilst no grade I actin embryos were noted. The addition of IGF-I resulted in the appearance of grade I actin embryos (8.33 and 6.9% for 10 and 100 ng/ml, respectively). These observations indicate that the addition of IGF-I during post-thaw culture can improve the quality of bovine cryopreserved embryos.
authors.affiliation: Institute for Cattle Breeding Ltd Rapotin, Vyzkumniku 267, 788 13 Vikyrovice, Czech Republic. makarevic@cvzv.sk
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Makarevich
authors.forename: Elena
authors.initials: E
authors.lastname: Kubovičová
authors.forename: Zdena
authors.initials: Z
authors.lastname: Hegedušová
authors.forename: Juraj
authors.initials: J
authors.lastname: Pivko
authors.forename: František
authors.initials: F
authors.lastname: Louda
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002452:Cell Count; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods*; D047108:Embryonic Development; D005260:Female; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D009028:Morula / Q000187:drug effects
chemical_list: D000199:Actins; D003470:Culture Media; D007334:Insulin-Like Growth Factor I