배양액에 첨가하는 에너지원이 생쥐 배 발생 능력에 미치는 영향

Effects of Energy Substrates in Culture Media on Developmental Capacity of Mouse Embryos

  • 박기상 (경북대학교병원 산부인과) ;
  • 이현정 (대구파티마병원 산부인과) ;
  • 박성백 (대구대학교 축산학과) ;
  • 김지철 (대구대학교 축산학과) ;
  • 이택후 (경북대학교병원 산부인과) ;
  • 전상식 (경북대학교병원 산부인과)
  • Park, Kee-Sang (Department of OB/GYN, Kyungpook National University Hospital) ;
  • Lee, Hyun-Jung (Department of OB/GYN, Daegu Fatima Hospital) ;
  • Park, Sung-Baek (Department of Animal Science, Daegu University) ;
  • Kim, Ji-Chul (Department of Animal Science, Daegu University) ;
  • Lee, Taek-Hoo (Department of OB/GYN, Kyungpook National University Hospital) ;
  • Chun, Sang-Sik (Department of OB/GYN, Kyungpook National University Hospital)
  • 발행 : 2007.03.31

초록

본 연구는 인간 난관액 또는 자궁액 내에 존재하는 에너지원이 생쥐 2-세포기 배의 체외 발달에 미치는 영향을 조사하기 위하여 실시하였다. ICR 암 생쥐에 5 IU hCG 주사 후 $46{\sim}50$ 시간에 2-세포기 배를 회수하였다. 회수된 배는 3가지 배양 조건 [대조군: 0 mM Group A: glucose(G) 0.5 mM + pyruvate(P) 0.32 mM + lactate(L) 10.5 mM, Group B: G 3.15 mM + P 0.1 mM + L 5.83 mM]에서 72시간 배양하였다. 배양 24 시간에 상실배 출현율은 group A (72.3%)와 group B (56.6%)가 대조군(34.9%)보다 유의하게 높았다.(p<0.05). 그러나 48시간에 배반포기 배 출현율은 대조군(51.8%)이 group A (39.8%)와 group B (25.9%)보다 유의하게 (p<0.05) 높았다. 72시간에 투명대 부착 ($ZiB,\;41.0{\sim}51.8%$), 투명대 탈출 ($ZeB,\;18.1{\sim}32.5%$) 및 총 배반포기 배 출현율 ($68.7{\sim}73.5%$)은 실험군 간에 통계적인 차이가 없었다. 배반포기 배의 평균 세포수와 ICM 세포수는 group A (70.8, 13.4)와 group B (64.4, 11.8)가 대조군 (53.1, 5.7)보다 유의하게(p<0.05) 많았고, 통계적인 유의차는 없었으나 group A가 group B보다 많은 경향이었다. 총 세포수에 대한 ICM 비율은 group A(22.9%)와 group B(23.7%)가 대조군(14.2%)보다 유의하게(p<0.05) 높았다. 영양배엽(TE) 세포수($34.1{\sim}45.1$)는 실험군 간에 통계적인 차이가 없었다. ICM에 대한 TE 비율(ICM:TE ratio)은 대조군(1:6.0)이 group A(1:3.4)나 group B(1:3.4)보다 유의하게(p<0.05) 높았다. 생쥐 2-세포기 배를 배양하여 72시간까지의 배 발달율을 살펴보면 배양액에 에너지원을 첨가하는 것이 효과적이었으며, 자궁액 농도보다는 난관액 농도로 에너지원을 조절했을 때 배 발생 능력이 높은 경향을 보였다.

This study was conducted to examine the effects of energy substrates in different conoentration of carbohydrates in the human oviduct and uterus on the in vitro development of mouse 2-cell embryos. Two-cell embryos were collected from ICR female mice at $46{\sim}50\;hr$ after 5 IU hCG injection and cultured in three different media [control: 0 mM, Guoup A: glucose (G) 0.5 mM + pyruvate (P) 0.32 mM + lactate (L) 10.5 mM, Group B: G 3.15 mM + P 0.1 mM + L 5.83 mM] for 72 hr. Rates of morula formation of group A (72.3%) and B (56.6%) were significantly higher higher (p<0.05) than that of control (34.9%) at 24 hr. However, blastocyst rate was significantly higher (p<0.05) in control (51.8%) than group A (39.8%) and B (28.9%) at hr. At 72 hr, no differences were found in the number of zona-intact, zona-escape and total blastocysts among groups. Mean and ICM cell numbers were significantly higher (p<0.05) in group A (78.0, 13.4) and B (64.4, 11.8) than control (53.1, 5.7), respectively, The percent of ICM were significantly higher (p<0.05) in group A (22.9%) and B (23.7%) than control (14.2%). No differences were round in the TE cell numbers ($34.1{\sim}45.1$). The ICM:TE ratio was significantly higher $34.1{\sim}45.1$ in control (1:6.0) than group A (1:3.4) and B (1:3.4). This study shows that energy substrates added to culture media especially, the oviductal level of carbohydrates increase the developmental capacity of 2-cell mouse embryos.

키워드

참고문헌

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