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한우 난포란의 채란방법에 따른 체외수정란의 생산효율
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  • 한우 난포란의 채란방법에 따른 체외수정란의 생산효율
  • Comparison of In Vitro Embryo Production with Follicular Oocytes Collected by Aspiration and Slicing in Korean Native Cows
저자명
이경미,곽대오,송상현,최양석,김윤연,강다원,하란조,윤창현,박충생
간행물명
韓國受精卵移植學會誌
권/호정보
1996년|11권 3호|pp.249-258 (10 pages)
발행정보
한국수정란이식학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

To improve the efficiency of in vitro production of embryos with follicular oocytes in Korean Native cows, the recovery rates, in vitro maturation, fertilization and development, and the time required for collecting and processing oocytes by aspiration with or without slicing were evaluated comparatively. The ovaries were obtained from a local abattoir and placed in physiological saline at 25~28$^{circ}C$ and brought to the laboratory within 3 hrs. The oocytes were collected by aspiration of follicles(2~6mm) with or without slicing ovaries after aspiration, and classified into Grade I, Grade II, Denuded, Expanded oocytes by the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules. Also the time required for each step of collecting and processing oocytes were measured. The cumulus cells were removed in some Grade I oocytes to measure their size and nuclear configuration before and after in vitro maturation. The Grade I oocytes were matured in vitro(IVM) for 24 hrs. in TGM-199 supplemented with 35$mu$g /ml FSH, 10$mu$g /ml LH, 1 $mu$g /ml at 39$^{circ}C$ under 5% C02 in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24hrs. and then the zygotes were cocultured in vitro (IVC) with bovine oviductal epithelial cells for 10 days. The results obtained were as follows: The number of oocytes recovered per ovary was averaged 6.6 by aspiration and 11.2 by slicing post aspiration, which summed to 17.8. The number of Grade I oocytes recovered per ovary was averaged 3.1 by aspiration and 3.6 by slicing, which summed to 6.7. The percentage of Grade I to total oocytes recovered was significantly(P<0.05) higher as 48.0 % in aspiration than 31.6% in slicing post aspiration. The time requlred for recovering a Grade I oocyte by aspiration and slicing was 1.1 and 2.5 min, respectively. The mean diameter of Grade I oocytes by aspiration and slicing was similar as 148.7 and 151.5$mu$m, respectively. The percentage of Metaphase II stage oocytes after IVM for 24 hours was significantly (P<o.05) lower in slicing(64.0%) than in aspiration(81.5%). Of the cleaved embryos, the percentage of embryos developed to blastocyst stage after culture for 10 days post insemination was similar as 23.3% and 20.5% in aspiration and slicing, respectively. These results suggest that the oocyte collection by slicing post aspiration could result in two fold increase in the yield of Grade I follicular oocytes and blastocysts from the bovine ovaries obtained at abattoirs, although the time required for collecting a Grade I oocyte was more than twice in slicing, compared with aspiration of oocytes from the visible follicles.