• Title/Summary/Keyword: 한우 체외수정란

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Production of In-Vitro Fertilized Embryos and Result of Transfer with Fresh or Frozen Embryos for Hanwoo and Holstein Cattle (한우 및 젖소에서 체외 수정란 생산과 신선 및 동결 수정란 이식 결과)

  • Kim Y. J.;Kim H. C.;Seo S. H.;Jeong K. N.;Kim Y. S.;Lee H. R.;Shin D. S.;Jo S. W.;Kim S. H.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.79-87
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    • 2005

  • The ovaries from Hanwoo and Holstein were collected from labattoir and transferred to laboratory. Oocytes were aspirated and incubated in $CO_2$ incubator for 24 hours for maturation. Oocytes were coincubated with the sperms for 5 hours. Cleaved oocytes were selected 48 hours after coincubation and half of the medium was changed newly every 48 hour until blastocyst formation. Cleavage rate and blastocyst rate were investigated according to different breeds and different status of cumulus cells surrounding the oocytes. Blastocysts were either transferred to the recipients or frozen until use. The result of embryo transfer with fresh or frozen embryos was investigated. The rate of male offspring following embryo transfer was also investigated. The rate of cleavage was $66.4\%$ for Hanwoo and $62.4\%$ for Holstein oocytes. The rate of cleavage according to status of oocyte was shown highest in the oocytes completely surrounded with cumulus cells and lowest in denuded oocytes for both Hanwoo and Holstein oocytes. The rate of blastocyst from cleaved oocytes was $40.6\%$ for Hanwoo and $36.9\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with fresh IVF embryos was $57.2\%$ for Hanwoo and $53.3\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with frozen IVF embryos was $40.9\%$ for Hanwoo and $36.4\%$ for Holstein. The rate of male calf produced by embryo transfer was $63.6\%$ for Hanwoo and $50.0\%$ for Holstein.

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Chromosomal Analysis of Hanwoo Embryos by In Vitro Culture Condition (한우 체외 수정란의 체외 배양 조건에 따른 염색체 분석)

  • Choi, S.H.;Cho, S.R.;Han, M.H.;Kim, H.J.;Choe, C.Y.;Son, D.S.;Chung, Y.G.;Kim, S.K.;Sohn, S.H.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.137-141
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    • 2007

  • Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

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무혈청 배양액을 이용한 한우의 체외수정란 생산

  • 정연길;석상현;박성백;임여정;최선호;송해범
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.137-137
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    • 2003

  • 난자의 체외성숙 및 체외배양에는 일반적으로 동물의 혈청을 기본배양액에 5-10% 정도 첨가한 배양액을 사용하고 있다. 그러나, 혈청으로부터 바이러스, 세균, 마이코 플라즈마 등에 오염될 가능성이 있기 때문에, 본 실험에서는 완전 무혈청 배양액에서 난자의 성숙, 배발생, 세포수, 동결성을 검토하였다. 도축된 한우의 난소로부터 채취한 난자는 선별하여 TCM199+10% FBS와 IVMD 101 배양액에서 22~24시간 동안 체외성숙시킨 후, IVF 100(일본, 펩티트연구소)으로 2회 세정한 후, 각각의 배양액 50${\mu}\ell$ 소적에 5개씩 5~6 시간 수정시켰다. 체외수정한 수정란은 TCM 199+10% FBS, IVMD 101, IVD 101 배양액에서 7~8일간 배양하여 배발생율을 조사하였다. 발생된 배반포의 일부는 세포수를 조사하였고 나머지 배반포는 1.8M EG로 동결하였다. (중략)

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흑염소의 체외수정란 생산

  • 최창용;한만희;권응기;최성복;최연호;최순호;손동수;노규진;최상용
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.143-143
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    • 2003

  • 흑염소는 우리나라 고유의 재래가축으로 다른 축종에 비해 아직까지 재래의 특성이 남아있는 가축이다. 또한 흑염소는 다른 경제동물에 비해 체구가 작고 온순하여 다루기 쉽고, 번식력이 좋으며, 임신기간이 짧은 장점을 가지고 있어 생명공학이나 형질전환동물의 생산과 같은 학문연구를 위한 실험용 동물로 많이 이용되고도 있다. 가축개량의 한 방법으로 오랫동안 수정란이식 기술이 연구ㆍ이용되고 있으나 한우, 젖소, 돼지 등의 한정된 동물에게만 연구ㆍ개발 되어왔으며, 흑염소에서는 이러한 연구가 미진한 실정이다. 이에 본 실험에서는 흑염소의 체외수정란 생산기술을 개발하여 생명공학실험을 위한 기초 동물로서 유용하게 활용하며 고유의 유전자원인 흑염소의 보존을 위한 기반을 마련하고자 한다. 체외수정란생산을 위해 사용된 난소는 도축장에서 도축된 재래 흑염소의 난소를 도축 즉시 적출하여 penicillin G와 streptomycin이 함유된 3$0^{\circ}C$내외의 생리식염수에 담아 1~2 시간내에 실험실로 운반하였다. 운반된 난소는 항생제가 첨가된 생리식염수로 3~4회 세척하여 실험에 공시하였다. (중략)

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난관상피세포의 체외배양시 IL-1$\alpha$ 및 $\beta$ 첨가에 의한 IGF-I의 생산

  • 최선호;조상래;한만희;김현종;손동수;류일선;김영곤;정영호;박춘근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.234-234
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    • 2004

  • 수정란이식 기술이 활성화되고, 한우의 가격이 상승하여, 젖소에 한우 체외수정란이식이 성행하고 있으나, 수정란이식에 의한 수태율은 수정란이식의 성과를 내고 있다고 할 정도는 아니다. 쥐, 돼지 및 사람에서 착상에 관련되는 인자에 대한 연구가 활발히 이루어지고 있으며, 가장 착상에 관여를 많이 하고 있는 인자는 LIF, IGF와 TGF-β인 것으로 보고되고 있다. 따라서 본 연구는 임신과 관련된 주요기관인 난관의 상피세포를 체외배양하고, 그때 착상을 유도하는 것으로 알려진 IL-1α와 β를 첨가하여 IGF-I의 생산을 조사하였다. (중략)

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Embryo Transfer with Sex-Determined Hanwoo Embryos Produced by In-vitro Fertilization (성감별된 한우 체외수정란의 수정란 이식)

  • 김용준;이창민;정구남;이해리;조성우;김용수;신동수;홍유미;유일정
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.97-108
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    • 2003

  • In-vitro fertilized Hanwoo embryos were biopsied for sex determination by PCR. Biopsied embryos were incubated for 1∼2 hours for the recovery. Those sexed Hanwoo embryos were transferred to 49 Hanwoo and 16 Holstein recipients from February 2000 to February 2001. Of 65 recipients, 14 cows(12 Hanwoo and 2 Holstein) delivered the same offspring as sex-determined by PCR, therefore the conception rate was 21.5%. 1. Total 65 embryos(male 35, female 30) were transferred to recipients, and 14 calves (male 6, female 8) were delivered. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex determination was 100.0%. 2. The conception rate after transfer with biopsied embryo between Hanwoo and Holstein was 24.5% and 12.5% 3. The conception rate after transfer with biopsied embryo between fresh and frozen-thawed embryos was 23.5% and 14.3%. 4. The conception rate according to the season when embryo was transferred was 11.8, 29.4, 23.5 and 20.0% for spring, summer, autumn and winter, respectively. 5. The conception rate according to embryo quality after biopsy was 41.7, 30.0 and 0.0% for excellent, good and fair quality. 6. The conception rate according to thickness of uterine horn was 71.4, 18.9, 11.8 and 0.0% for 0, +, ++ and +++ thickness. 7. The conception rate according to the site in the uterine hem where embryo was put was 30.0, 20.0 and 10.0% for cranial, mid, and caudal part of uterine horn. 8. The conception rate according to the quality of corpus luteum ipsilateral to the uterine horn where embryos was transferred was 41.2, 14.3 and 15.4% for excellent, good and fair quality. 9. The conception rate according to the time required for embryo transfer was 18.2, 30.0, 30.0, 0.0 and 25.0% for 10, 15, 20, 25 and 30 minutes. 10. The conception rate according to parity of recipients was 26.5, 19.1, 14.3 and 0.0% for the primiparous, the 2nd parous, the 3rd parous and the 4th parous recipients. These results indicated that fresh embryos are more demanded than frozen-thawed embryos for good conception rate in embryo transfer with biopsied-sexed embryo. Also, it was indicated that we should consider embryo-recovering condition, recipient's uterine thickness, transfer site in uterine horn, quality of corpus luteum, time required for transfer and parity of recipient to achieve good conception rate in ET with biopsied-sexed embryos.

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