본 연구는 소 체외성숙, 체외수정 유래 배반포의 초자화 등결 시 동해방지제의 평형방법과 응 해 후 동해방지제의 희석방법이 수정란의 생존성에 미치는 영향을 검토하고자 실시하였다. 초자화 동결액은 20% FBS(Gibco)가 첨가된 D-PBS에 20% glycerol, 20% ethylene glycol, 3/8 M sucrose, 3/8 M deftrose 가 함유된 GESD를 사용하였다. 동결 전 평형방법은 3단계 (El), 2단계 (E2), 1단계(E

The purpose of this experiment was to determine the effects of thiol compounds, -mercaptoethanol(-ME) and cystearrone with buffalo rat liver cell(BRLC) co-culture on the development and intracellular glutathione(GSH) concentrations of bovine embryos produced by in vitro inaturation(IVM) and in vitro fertilization(IVF). Bovine IVM /IVF embryos developed to 2~8 cell stage were co-cultured with BRLC in GRlaa with or without thiol compounds. The developmental rate beyond morulae stage in CRlaa containing 0, 10,25 and 50M -ME with BRLG were 63.0, 74.0, 72.3 and 77.1%, respectively. And the developmental rate with 0, 25, 50 and 75M cystearnine with BRLC were 69.6, 77.6, 81.0 and 76.8%, respectively. The developmental rate beyond morulae stage of GRlaa containing thiol compound with BRLG group was higher than that of control group. The intracellular GSH concentrations of blastocysts cultured for 5 days in GRlaa containing 0 and 50M -ME or cysteamine with BRLG were 81.2 and 86.4, 83.2 and 84.2pM, respectively. The intracellular GSH concentrations of blastocysts in GRlaa containing thiol compounds with BRLG was slightly higher than that of control group The cell numbers of blastocysts were not difference in all experimental groups. These results indicate that thiol compounds with BRLG co-culture was increased the percentage of developed into morulae and blastocysts, and intracellular GSII concentrations of blastocysts embryos.

The objective of this study was to investigate the effects of thiol compounds with bovine oviduct epithlial crlls(BOEC) co culture on development and intracellular glutathione(GSH) concentrations of bovine embryos derived from IVM /IVF oocytes. In experiment 1 and 2, embryos developed to 2~8 cell stage after in vitro fertilization were co-cultured with BOEC in CRaa with or without -mercaptoethanol(-ME) and cysteamine. The percentage of embryos that developed to morulae and blastocysts in 0,10, 25 and 5OM -ME with BOEC was 48.1, 64.0, 72.9 and 75.9%, respectively. Twenty-five and 5OM -ME groups were significantly higher than in 0 and 1OM - -ME groups(PM cysteamine with BOEC was 50.0, 53.2, 72.0 and 66.7%, respectively. Fifty M cysteamine group was significantly higher than any other groups (Paa with 0 and 5OM -ME or cysteamine were 68.5, 77.8, 78.7 and 80.0pM, respectively. Fifty M -ME group was significantly higher than that of control(P<0.05), but cysteamine group was not. Cell numbers of blastocysts were not difference in all experimental groups. These experiments indicate that -ME and cysteamine with BOEC co-culture can affect the development and intracellular GSH concentrations of bovine embryos produced by IVM /IVF docytes.