Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Effects of Energy Substrates in Culture Media on Developmental Capacity of Mouse Embryos

배양액에 첨가하는 에너지원이 생쥐 배 발생 능력에 미치는 영향

  • 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)
  • 박기상 (경북대학교병원 산부인과) ;
  • 이현정 (대구파티마병원 산부인과) ;
  • 박성백 (대구대학교 축산학과) ;
  • 김지철 (대구대학교 축산학과) ;
  • 이택후 (경북대학교병원 산부인과) ;
  • 전상식 (경북대학교병원 산부인과)
  • Published : 2007.03.31
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Abstract

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.

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.

Keywords

References

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