Abstract
Three experiments were performed to develop an optimal culture system of in-vitro derived porcine embryos. In experiment 1, embryos were cultured in two different basic culture media (NCSU-23 and PZH) supplemented with BSA (4mg/ml), FBS (10%), PVA (3mg/ml). Although no difference on the percentage of embryos developed to blastocysts was found (P>0.05), more 2-cell embryos (59.5% vs. 44.7 to 52.0%) were obtained when embryos were cultured in NCSU-23 added with BSA. In experiment 2, identical treatment was conducted using PZM medium. More (P<0.05) 2-cell embryos (54.4% vs. 40.7 to 44.5%) and blastocysts (12.5% vs. 5.8 to 9.0) were produced when embryos were cultured in PZM added with BSA (4mg/ml). In experiment 3, embryos were cultured in media as follows; 1) NCSU-23 without BSA for 192 hours; 2) NCSU-23 with BSA for 192 hours; 3) NCSU-23 with BSA for 96 h and then subsequently injected with FBS into culture drops. Although no difference on the percentage of embryos developed to blastocysts was found (P>0.05), more 2-cell embryos (63.8 to 69.2% vs. 52.0%) were obtained when embryos were cultured both in NCSU-23 added with BSA throughout culture duration and in FBS supplemented medium. In conclusion, no difference was found when porcine preimplantation embryos were cultured in NCSU-23 added with various macromolecules. However, when PZM medium was used, PZM medium supplemented with BSA showed more potential to support embryo development. Finally, FBS supplemented into culture drop 96h post-insemination did not show any benefits on embryo development.
본 연구에서 배양액 내 첨가된 macromolecule이 돼지 체외수정란의 체외 발달율에 미치는 영향과 배양소적 내 첨가된 FBS가 후기배발달에 미치는 영향에 대하여 알아보았다. 이에 대한 결과를 요약 하면 다음과 같다. 1. NCSU-23을 기본 배양액으로 사용시 BSA (4mg/ml), FBS (10%), PVA (3 mg/ml)가 각각 첨가되었을 경우 배발달에 큰 차이를 보이지 않았다. 2. PZM을 기본 배양액으로 사용시 BSA (4mg/ml)가 첨가하였을 경우 2-세포 수정란 발달율과 배반포 발달율이 가장 높았으며, 이것은 NCSU-23에 비하여 PZM 배양액에는 후기배의 에너지원으로 사용하는 글루코스가 포함되어 있지 않기 때문으로 사료 된다. 본 연구를 통하여 돼지 수정란의 체외배양에 사용되는 배양액은 종류와 조성에 따라 적절한 macromolecule이 첨가되어 초기배와 후기배의 발달을 모두 촉진시킬 수 있어야 함을 알 수 있었다.