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체외 배양시 과립막세포와 공배양된 돼지 난포란의 성숙과 수정
박수봉,박항균,입곡명 ( S . B . Park,H . K . Park,Akira Iritani ) 한국축산학회 1990 한국축산학회지 Vol.32 No.1
The present study was undertaken to examine the maturation and fertilization of porcine oocytes cocultured with granulosa cells. The results obtained were as follows. The percentages of oocytes reaching metaphase Ⅱ by GC -free culture and coculture with MGC and LGC were 32, 28 and 24% at 33h, respectively. In the culture from 33h to 42h, rates of oocyte maturation showed no difference among experimental groups. The fertilization rates of oocytes matured by GC-free culture and coculture with MGC and LGC were 92, 97 and 94%, respectively, Male pronucleus formation rates of oocytes matured by GC-free culture and coculture with MGC and LGC were 49, 55 and 78% respectively and markedly increased by coculture with LGC. The present results suggest that coculture with the membrana granulosa cells does not affect the incidence of maturation, and that pronucleus formation was promoted through coculture with LGC.
박수봉,박항균,입곡명 ( S . B . Park,H . K . Park,Akira Iritani ) 한국축산학회 1990 한국축산학회지 Vol.32 No.1
The present study was conducted to investigate the optimum conditions of in vitro fertilization by ejaculated boar spermatiozoa capacitated in a defined medium. When fresh ejaculated boar spermatozoa were preincubated for 4h with elevated sperm concentration fertilization rates were markedly increased. The fertilization rates by spermatozoa preincubated at the concentrations of 2, 4, 10, 20 and 40×10^8 cells/ml were 12, 30, 50, 76 and 82%, respectively, However, when spermatozoa preserved for 10-12h at 20℃ were preincubated for 4h with elevated sperm concentration fertilization rates were markedly decreased. The fertilization rates by spermatozoa preincubated at the concentrations of 2, 4, 10, 20 and 40×10^8 cells/ml at preincubation were 91, 77, 68, 44 and 8%, respectively. When oocytes matured in vitro were inseminated with spermatozoa at the concentration 1 × I0^6 cells/ml, rates of fertilization and polyspermy were 97 and 87%, respectively, after culture for 20h without changing medium. When oocytes were inseminated with spermatozoa at the different concentrations of 1, 0.5, 0.25 and 0.1×10^6 cells /ml and the medium was changed 5h after insemination, the fertilization rates were 75, 73, 61 and 30%, respectively. The rates of polyspermic fertilization were 58, 47, 40 and 47%, respectively, when oocytes were inseminated with different concentrations of spermatozoa, and there was no difference in the rates of polyspermy fertilization among groups with changing medium. The present results suggested that both sperm preincubation with high concentration and sperm preservation at 20℃ for 10-12h were effective for capacitation ejaculated boar spermatozoa and that a complete block of polyspermy was not achieved in this in vitro fertilization conditions.
돼지 난포란의 성숙과 수정에 대한 성숙배지에 첨가된 난포액의 영향
박수봉,박항균,입곡명 ( S . B . Park,H . K . Park,Akira Iritani ) 한국축산학회 1990 한국축산학회지 Vol.32 No.2
The purpose of these experiments were to investigate the effect of follicular fluid supplemented to maturation medium on the maturation and fertilization of porcine follicular oocytes. The results obtained were as follows. In the media with MFF and LFF, the rates of maturation division of porcine oocytes in vitro were 82% and 80%, respectively, and the proportion of oocytes matured to metaphase Ⅱ become nearly constant after 39h of culture. However, the rates of maturation of porcine oocytes in medium with FCS 10% were 60% and 79% at 39 and 42h, respectively. The fertilization rate of oocyte cultured in the medium with FCS 10% is 99%. When the oocytes matured in medium with MFF and LFF were fertilized, fertilization rates were 42% and 54,%, respectively. The male pronucleus formation rates of oocytes cultured in the media with FCS 10% MFF 10% and LFF 10% concentrations were 54, 74 and 84%, respectively and were increased through culture in media supplemented with follicular fluid. The present results clearly demonstrate that follicular fluid promoted the maturation of oocytes and male pronucleus formation.