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Vitrification of oocytes and in vitro produced bovine embryos exposed to cytochalasin B

Vitrificação de oócitos e embriões bovinos produzidos in vitro e expostos à citocalasina B

In this study, the influence of cytochalasin B on the survival and development after vitrification of oocytes and in vitro produced (IVP) bovine blastocysts was evaluated. In the Experiment I, 956 oocytes were matured for 22h and were immediately vitrified (Vitri treatment) or exposed for 15-20 minutes, to 7.5µg/ml (CB7.5Vitri treatment) or 45µg/ml (CB45Vitri treatment) cytochalasin B solutions, before vitrification. After 30 seconds of exposure to SV1 [400µl TCM-Hepes with 10% fetal serum (SF), 50µl ethylene glycol (EG) and 50µl DMSO], and 20 seconds to SV2 (300µl trehalose 1,0M + 20% SF, 100µl EG and 100µl DMSO) solutions, the oocytes were vitrified in Open Pulled Straws (OPS). The rewarming was performed at 37-38ºC in two steps of 5 minutes each, into 0.3 and 0.15M trehalose solutions, respectively. There was no difference (P>0.05) in the cleavage and embryo rates between Vitri, Cito7.5Vitri and Cito45Vitri treatments, which were inferior to control group (P<0.05). In the Experiment II, 269 expanded blastocysts were distributed into three treatments. In the Vitri treatment blastocysts were exposed for 1 minute to SV1 and for 20 seconds to SV2 solution, the later one modified using TCM-Hepes instead of trehalose. In the CB45Vitri treatment, embryos were vitrified after being exposed to a 45µg/ml cytochalasin B solution for 10 minutes. The re-expansion and hatching rates were similar to Vitri and CB45Vitri treatments (P>0.05), which were lower than those observed in the Control group (P<0.05). The results show that, independently of dose studied (7.5 or 45mg/ml), cytochalasin B has not a beneficial effect to the vitrification of oocytes and IVP bovine blastocysts.

Bovine; Cytochalasin B; Embryos


Faculdade de Medicina Veterinária e Zootecnia / Universidade de São Paulo Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária Armando de Salles Oliveira, 05508-270 São Paulo SP Brazil, Tel.: +55 11 3091-7636, Fax: +55 11 3031-3074 / 3091-7672 / 3091-7678 - São Paulo - SP - Brazil
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