title: Effect of vitrification medium composition and exposure time on post-thaw development of buffalo embryos produced in vitro.
abstract: This study examined the effects of different vitrification medium compositions and exposure times (2, 4 and 6min) on the post-thaw development of buffalo embryos produced in vitro (IVP). The compositions were (1) 40% ethylene glycol (EG); (2) 25% glycerol (G)+25% EG, and (3) 25% EG+25% dimethylsulfoxide (DMSO). The base medium was 25mM Hepes-buffered TCM-199+10% steer serum +50microg/mL gentamycin. The IVP embryos were cryopreserved by a two-step vitrification method at 24 degrees C. After warming, the embryos were cultured in vitro for 72h. The vitrification of morulae and blastocysts in 25% EG+25% DMSO with an exposure time of 2 and 4min, respectively, resulted in a better hatching rate than other combinations. The hatching rate of morulae vitrified in 25% EG+25% G, 25% EG+25% DMSO, and blastocysts vitrified in 40% EG, 25% EG+25% DMSO were negatively correlated with exposure time. However, the hatching rate of blastocysts vitrified in 25% EG+25% G was positively correlated with exposure time. The study demonstrated that the post-thaw in vitro development of IVP buffalo embryos was affected by the vitrification medium composition and exposure time.
authors.affiliation: Department of Animal Reproduction, Veterinary College, Karnataka Veterinary, Animal and Fishery Sciences University, Hebbal, Bangalore 560 024, India. drmanjunathvet@gmail.com
authors.forename: B M
authors.initials: BM
authors.lastname: Manjunatha
authors.forename: P S P
authors.initials: PS
authors.lastname: Gupta
authors.forename: J P
authors.initials: JP
authors.lastname: Ravindra
authors.forename: M
authors.initials: M
authors.lastname: Devaraj
authors.forename: S
authors.initials: S
authors.lastname: Nandi
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000196:embryology* / Q000502:physiology; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D003470:Culture Media / Q000737:chemistry; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D005990:Glycerol / Q000494:pharmacology; D008297:Male; D013997:Time Factors
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media; D019855:Ethylene Glycol; D005990:Glycerol; D004121:Dimethyl Sulfoxide