• Journal of Embryo Transfer
• /
• v.12 no.1
• /
• pp.91-102
• /
• 1997

This study was carried out to enhance the efficiency of Korean Native cattle embryos and establish the techniques for producing the twin calves. Bisected embryos without zona pellucida which were divided by simple method not using holding pipette or whole two embryos were transferred to recipients.The pedigrees of monozygotic twin calves produced by transfer of bisected pair embryos were identified. The results obtained were as follows ; The average successful bisection rate was 89.16%. The embryos of blastocyst stage (91.66%) were bisected successfully at significantly (P<0.05) higher rate, compared with the morula stage embryos (86.66%). The average survival rate of bisected embryos following 24 hours culture was 59.02%. The survival rate of morula stage embryos (62.50%) was significantly (P<0.05) higher than that of blastocyst stage embryos (55.5%). For the production of monozygotic twin calves, ten pairs of flesh or frozen demi-em- lymphocytes antigen, the twin calves produced by transfer of bisected pair embryos of Korean Native cattle were identified in pedigrees and confirmed as monozygotes.

• Journal of Embryo Transfer
• /
• v.12 no.3
• /
• pp.325-333
• /
• 1997

수정란이식의 주변기술인 초기배의 미세조작및 성 판정은 가축의 경우, 경제 형질의 유전적 개량에 크게 기여하였다. 본 연구는 미세조작 biopsy와 PCR에 의한 체외생산 소 초기배의 급속. 정확한 성 판정 기법을 확립하기 위해 실시하였다. 체외성숙 및 체외수정에 의해 생산된 소수정란은 소 난관상피세포와 공배양을 통해 8-세포부터 배반포시시까지 체외발생시킨 후 미세조작 biopsy에 이용되었다. 미세조작 biopsy 과정중에 약간의 형태적인 손상이 관찰되었지만 대부분의 demi-embryo는 정상적인 배반포와 나화배반포로 발생하였다. 8~16 세포, 상실배, 초기배반, 포 시기에서 미세조작 biopsy 후 확장 또는 나환배반포기시기까지의 발달율은 각각 62.8(27/43), 83.3(30/36) 및 80.9%(55/68)로 정상적인 초기배와 큰 차이가 없는 것으로 나타났다. 총 136개의 소 초기배로 부터 2~10개의 할구세포를 미세조작 biopsy에 의해 분리하여 소 특이와 Y-특이염기서열을 가진 두 쌍의 염기서열을 이용하여 PCR을 수해하였다. 이들중 112(82.4%)개를 성공적으로 성 판정하였으며, 암/수 비율은 각각 34.8(39/112)/65.2%(73/112)로 나타났다. 본 실험에서 얻은 결과는 확립된 소 초기배의 미세조작기술과 성 판정 방법을 통해 계획적인 암.수 송아지의 선별 생산뿐만 아니라 소 수정란 이식을 통한 유전적 개량을 촉진시키는데 효과적으로 이용될 수 있는 기술이 확립된 것을 보여주었다.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2007.05a
• /
• pp.39-50
• /
• 2007

Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.

• Journal of Veterinary Clinics
• /
• v.7 no.2
• /
• pp.511-515
• /
• 1990

Induction of twinning was attempted by transfer of two whole- or demi-embryos in Holstein cows and heifers. Cows were superovulated with follicle stimulating hormone(FSH) administered twice daily in intra-muscular injection. On day of 6.5 to 7 post-estrus, embryos were collected nonsurgically, Normal morulae and early balstocysts were obtained from superovulated cows. The embryos were hisected with a micro-blade made from a razor. Twenty seven pairs of half embryos were successfully produced and 23 pairs of half embryos were transferred to recipients( Group A). Twenty cows were treated with low unit of FSH (3mg${\times}$2,2mg${\times}$2 ; Group B1, 2mg${\times}$2, 1mg${\times}$2, 1mg${\times}$2 ; Group 32) to induce double or triple ovulations. The cows of Group B1 and B2 were Inseminated artificially(A1) at following estrus. Twenty four heifers were bred by A1 and received an additional embryo into the uterine horn contralateral to the corpus luteum(CL) 6 days later(Group C). One embryo was transferred into each uterine hem of 16 heifers 6 days after estrus(Group D). Fourteen heifers were received two embryos into the uterine horn ipsilateral to the CL at day 6 of estrous cycle(Group I). Pregnancy rates at 60 days in Group A, B1, B2, C, D and E were 34.8％, 70.0％, 60.0％, 66.7％, 62.5％ and 57.1％, respectively. Twinning rates were 8.7％, 20.0%, 10.0％, 16.7％, 18.7％ and 21.4％, respectively. The present experiments demonstrate that FSH treatment( Group 31) and ipsilateral transfer( Group I ) of two whole-embryos are more useful methods to produce twins than the others.