title: Prolonging bovine sperm-oocyte incubation in modified medium 199 improves embryo development rate and the viability of vitrified blastocysts.
abstract: This study was conducted to compare the efficacy of four in vitro fertilization (IVF) media: Bracket and Oliphant's medium (BO), modified medium 199 (IVF-M199), modified Tyrode's medium (MTM), and modified KSOM (m-KSOM) on fertilization efficiency and blastocyst formation rate. In addition, we wanted to investigate the benefit of prolonging the IVF period (from 6 to 18 h) using the two most effective IVF media determined in our initial experiment; subsequently, blastocyst viability was assessed following vitrification. A higher incidence of polyspermic fertilization was observed in the MTM (6%) and in BO, in both the 6 and 18 h (7% and 11%, respectively) groups, than in the m-KSOM (1%) or in the IVF-M199 6 or 18 h (1 and 3%, respectively) groups. Cleavage rates were similar in BO, IVF-M199, and MTM 48 h post-fertilization; however, the lowest cleavage rate was observed for m-KSOM. A greater proportion of zygotes developed into 8-cell embryos in IVF-M199 than in other IVF media. Subsequently, a greater proportion of blastocyst formation and hatching was achieved in IVF-M199 (40% and 79%, respectively) or BO (35% and 74%, respectively) than in m-KSOM (18% and 58%, respectively) or MTM (22% and 66%, respectively). Prolonging IVF to 18 h did not alter cleavage rates; however, the highest rate of overall blastocyst formation was achieved in the IVF-M199 18 h (49%), rather than in the BO 18 h (20%) group. Vitrified/thawed blastocysts from IVF-M199 groups re-expanded and developed better, as compared to the BO 18 h group, and hatching rate and total cell number in IVF-M199 18 h group was comparable to the control groups (non-vitrified). Vitrification reduced survival compared to controls. In conclusion, IVF-M199 was successfully used for IVF, compared favorably to BO medium, and offered the advantage of an extended IVF period for up to 18 h that requires only one-half a dose of semen, and resulted in better quality blastocysts that endured vitrification with a hatching rate comparable to that of control groups.
authors.affiliation: Center for Regenerative Biology/Department of Animal Science, University of Connecticut, 1392 Storrs Rd., U-4243, Storrs, CT 06269, USA.
authors.forename: T L
authors.initials: TL
authors.lastname: Nedambale
authors.forename: F
authors.initials: F
authors.lastname: Du
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: S A
authors.initials: SA
authors.lastname: Chaubal
authors.forename: A
authors.initials: A
authors.lastname: Dinnyes
authors.forename: W
authors.initials: W
authors.lastname: Groen
authors.forename: D
authors.initials: D
authors.lastname: Faber
authors.forename: J R
authors.initials: JR
authors.lastname: Dobrinsky
authors.forename: X
authors.initials: X
authors.lastname: Yang
authors.forename: X C
authors.initials: XC
authors.lastname: Tian
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D004624:Embryo Transfer / Q000662:veterinary; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000592:standards / Q000662:veterinary*; D008297:Male; D013997:Time Factors; D014022:Tissue Survival
chemical_list: D003470:Culture Media