Journal of Veterinary Clinics (한국임상수의학회지)
- Volume 12 Issue 1
- /
- Pages.877-886
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- 1995
- /
- 1598-298X(pISSN)
- /
- 2384-0749(eISSN)
Study on Production of Cloned Animals by Recycling Nuclear Transplantation III. Production of Third Generation Cloned Embryos in Rabbits
반복핵이식에 의한 복제동물 생산에 관한 연구 III. 토끼에서 제3세대 복제수정란의 생산
- Lee Hyo-jong ;
-
Jeon Byeong-gyun
;
- Yin Xi-jun ;
-
Park Choong-saeng
;
-
Choe Sang-yong
;
- Yun Chang-hyun ;
- Kang Dae-jin
- 이효종 (경상대학교 축산진흥연구소) ;
-
전병균
(경상대학교 축산진흥연구소) ;
- 윤희준 (경상대학교 축산진흥연구소) ;
-
박충생
(경상대학교 축산진흥연구소) ;
-
최상용
(경상대학교 축산진흥연구소) ;
- 윤창현 (경상대학교 축산진흥연구소) ;
- 강대진 (경상대학교 축산진흥연구소)
- Published : 1995.06.01
Abstract
The recycling nuclear transplantation(NT) technique has the powerful potential of producing a large number of genetically identical embryos and offsprings from one embryo. Multiple generational cloning by this technique utilizes the NT embryo itself as the donor for the next generation of cloning. In this experiment, we have produced the third generational cloned embryos by recycling NT. Further we examined comparatively the electrofusion rate and in vitro developmental potential in the cloned embryos of the first second and third generations. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulberco's phosphate buffered saline containing 10 % fetal calf serum(FCS) at 47 hours after hCG injection. In the first generation NT, the nuclear donor embryos were synchronized in the phase of Gl/S transition of 32-cell stage. The first and second generation NT embryos developed to 16-cell were used as donor nuclei for second and third generation. The recipient cytoplasms were utilized the oocytes collected at 14 hours after hCG injection, following revoming the nucleus and the first polar body by micromanipulation. The separated blastomeres were injected into the enucleated recipient oocytes by micromanipulation and were fused by electrical stimulation. The electrofusion rate was seen to be 78.0, 88.0 and 90.3 % in the first second and third generation NT rabbit embryos, respectively. The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10 % FCS for 120 hours at 39