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돼지체외수정 시 Methyl-beta-cyclodextrin이 정자 첨체반응과 분할율에 미치는 영향

Effect of Methyl-beta-cyclodextrin on Sperm Acrosome Reaction and Embryo Cleavage after in vitro Fertilization in Pig

이용승|박춘근|정희태|양부근|박솔
동물생명과학연구소 (Institute of Animal Life Science)|2009.12|동물자원연구 (ANNALS OF ANIMAL RESOURCE SCIENCES)|제20권 |pp. 6-12

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참고문헌

키워드

CaffeineIn vitro fertilizationPigMethly-beta-cyclodextrinAcrosome reactionEmbryo cleavage
주제분류축산학

초록

This study investigated the effect of methly-beta-cyclodextrin (MBCD) on sperm acrosome reaction and embryo cleavage after in vitro fertilization in pig. The acrosome reaction test of pig sperm were evaluated by Coomassie Brilliant Blue (CBB) stain after incubatation (39℃, 5% CO2) in mTBM. Control group was fertilized without treatment reagent. Treatment groups were fertilized with caffeine, MBCD and caffeine with MBCD for 0, 30, 60, 180 and 360 min. Porcine oocyte-cumulus complexes were cultured in TCM-199 medium containing 10% porcine follicular fluid (pFF) and hormonal supplement with FSH, LH, hCG and EGF for 21~22 hrs. They were then cultured in the same medium without hormonal supplement for additional 21~22 hrs. The matured oocytes were denuded by 0.1% hyaluronidase treatment. The oocytes were in vitro fertilized with 1.5×105/㎖ sperm concentration in four different mTBM medium not contained(control) and contained caffeine, MBCD or caffeine+MBCD for 6 hrs. After in vitro fertilization, embryos were transferred into PZM-3 with 0.03% BSA medium. The cleavage of embryo was evaluated at 48 and 96 hrs. Results showed that when the sperm was incubated in four different mTBM medium, the integrity of acrosome was significantly (p<0.05) lower treated with MBCD than other treatment groups at 0, 30 and 360min. The cleavage rates of pig embryos was significantly (p<0.05) lower in control than other treatment groups, However, It was not significantly different between groups. In conclusion, the MBCD treatment in pig sperm showed effect on acrosome reaction, but cleavage rate of embryo was not affected.

Abstract

This study investigated the effect of methly-beta-cyclodextrin (MBCD) on sperm acrosome reaction and embryo cleavage after in vitro fertilization in pig. The acrosome reaction test of pig sperm were evaluated by Coomassie Brilliant Blue (CBB) stain after incubatation (39℃, 5% CO2) in mTBM. Control group was fertilized without treatment reagent. Treatment groups were fertilized with caffeine, MBCD and caffeine with MBCD for 0, 30, 60, 180 and 360 min. Porcine oocyte-cumulus complexes were cultured in TCM-199 medium containing 10% porcine follicular fluid (pFF) and hormonal supplement with FSH, LH, hCG and EGF for 21~22 hrs. They were then cultured in the same medium without hormonal supplement for additional 21~22 hrs. The matured oocytes were denuded by 0.1% hyaluronidase treatment. The oocytes were in vitro fertilized with 1.5×105/㎖ sperm concentration in four different mTBM medium not contained(control) and contained caffeine, MBCD or caffeine+MBCD for 6 hrs. After in vitro fertilization, embryos were transferred into PZM-3 with 0.03% BSA medium. The cleavage of embryo was evaluated at 48 and 96 hrs. Results showed that when the sperm was incubated in four different mTBM medium, the integrity of acrosome was significantly (p<0.05) lower treated with MBCD than other treatment groups at 0, 30 and 360min. The cleavage rates of pig embryos was significantly (p<0.05) lower in control than other treatment groups, However, It was not significantly different between groups. In conclusion, the MBCD treatment in pig sperm showed effect on acrosome reaction, but cleavage rate of embryo was not affected.

참고문헌 (39)

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