• Title/Summary/Keyword: 체외 수정

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넙치, Paralichthys olivaceus의 정자형성에 관한 미세구조적 연구

  • Kim, Jae-Won;Lee, Jung-Sik;Jin, Pyung
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.10a
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    • pp.252-253
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    • 2000
  • 어류에서 정자의 형태 및 구조에 관한 연구는 진화 유연관계 추적을 위해 그들의 변이성을 토대로 행해지고 있다. 어류 정자의 형태 및 미세구조는 종간 또는 그들의 생식생태와 관련하여 그 변이가 아주 다양하다. 일반적으로 어류의 정자는 수정 장소에 따라 체외수정형(aquasperm type)과 체내수정형(introsperm type)으로 나뉘고, 이는 다시 첨체의 유무에 따라 첨체형(acrosomal type)과 무첨체형(anacrosomal type)으로 나누어진다(Jamieson, 1991). (중략)

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Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.215-223
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    • 2003
  • This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Study on In Vitro Fertilization of Pig Oocytes Matured in In Vivo and In Vitro (생체(生體) 및 실험실(實驗室)에서 성숙(成熟)된 돼지 난모세포(卵母細胞)의 체외수정(體外受精)에 관(關)한 연구(硏究))

  • Park, Chang Sik;Lee, Kyu Seung;Seo, Kil Woong
    • Korean Journal of Agricultural Science
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    • v.18 no.2
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    • pp.114-118
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    • 1991
  • This study was conducted to find out the suitable in vitro media for fertilization and culture on the pig oocytes matured in vivo and in vitro. The results obtained were as follows; A comparison of fertilization media on the oocytes matured in vivo was made between M199 with 10% FCS versus TL Hepes with 1% BSA Fertilization rate was significantly higher in the TL Hepes medium. But polyspermic incidence did not favor in the TL Hepes medium. Embryos were cultured in TL Hepes wash and culture medium for 48h after insemination. A total of 53 embryos were cultured and 39(73.6%) cleaved. Of these 39 embryos, 31(79.5%) cleaved equally to the 2-8 cell stage. Immature oocytes cultured in Waymouth's maturation medium were much more able to induce sperm swelling than immature oocytes cultured in TL Hepes maturation medium, however, most ova were polyspermic and did not develope to the 4 cell stage during 48h culture in TL Hepes wash and culture medium.

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Outcome of in vitro Fertilization in Women $\geq$40 Years of Age (40세 이상의 여성에서 연령에 따른 체외수정시술 결과 분석)

  • Kwon, Cheo-Jin;Lee, Sang-Hee;Kang, Eun-Hee;Kim, Jong-Sik;Jung, Byeong-Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.285-291
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    • 2007
  • Objective: To evaluate the outcomes of in vitro fertilization and embryo transfer (IVF-ET) in women over 40 years of age. Methods: A total of 170 patients (271 cycles) over 40 years of age who underwent IVF-ET at Seoul Women's Hospital (Incheon, Korea) were analyzed in this study retrospectively. The patients were grouped into the women <44 years old group and the women $\geq$44 years old group. Statistical analysis was performed using Student's t-test and Fisher's exact test as appropriate. Results: An overall clinical pregnancy rate per retrieval was 11% (30/271). Of these, clinical miscarriage rate were 33% (10/30) and the overall delivery rate was 7.4% (20/271) per retrieval, respectively. The women $\geq$44 years old group had significantly higher cancellation rate (13% vs. 25%), lower number of retrieved oocytes (6.17$\times$4.62 vs. 4.13$\times$4.07), decreased number of 2PN (4.83$\times$3.61 vs. 3.46$\times$3.12), and reduced embryos for transfer (3.52$\times$1.72 vs. 2.81$\times$1.83) than the women <44 years old group. We found significantly lower clinical pregnancy rate (13.0% vs. 2.1%) and live birth rate (9.0% vs. 0.0%) in the women $\geq$44 years old group than the women <44 years old group. Conclusion: The present study has shown that IVF outcome is seriously impaired in the women $\geq$44 years old.

생명공학 기술 적용을 위한 닭 수정란의 체외 배양법

  • 전익수;박진기;이연근;노환국;최철환;김학규;이지현;김선화;이필헌
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.45-45
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    • 2002
  • 닭 수정란을 이용한 생명공학 기술은 일반적인 포유류 가축의 수정란에 적용되고 있는 기술과 비교 해 볼 때, 기본적인 배 발생 단계에서부터 개체 탄생에 이르기까지 상당 부분에 걸쳐 상이한 점이 많다. 왜냐하면, 조류는 발생학적 특성상 수정과 초기 배 발생을 제외한 거의 대부분의 개체 발생 과정이 난각 속에서 진행되기 때문이다. 또한, 포유류는 새끼를 낳는 반면, 조류는 몸밖에 알을 낳아 적당한 습도와 온도에서 일정기간 품은 다음 병아리를 부화시키는 동물이다. (중략)

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Effect of Testicular Histopathology on Pregnancy Outcomes in Non-Obstructive Azoospermia (비폐쇄성 무정자증 환자에서 고환의 조직병리학적 진단에 따른 체외수정시술 결과의 비교)

  • Park, Chan-Woo;Seo, Ju-Tae;Park, Yong-Seog;Kim, Hye-Ok;Yang, Kwang-Moon;Kim, Jin-Young;Koong, Mi-Kyoung;Kang, Inn-Soo;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.293-301
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    • 2008
  • Objective: To evaluate outcomes of patients with non-obstructive azoospermia (NOA) undergoing the testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection (ICSI) with different histopathologic subgroups. Method: A total of 122 embryo-transferred TESE/ICSI cycles were compared among NOA subgroups; Germ-cell aplasia (GA, 40 cycles), Maturation arrest (MA, 32 cycles) and severe hypospermatogenesis (S-HS, 50 cycles). Obstructive azoospermia (OA, 667 cycles) patients were served as a control. TESE/ICSI outcomes such as fertilization rate (FR), clinical pregnancy rate (CPR) and live birth rate (LBR) were evaluated. Results: The 2PN FR of embryo-transferred TESE/ICSI cycle was 58.1% in GA, 42.2% in MA and 48.0% in S-HS, which was significantly lower than that of OA (72.9 %, p<0.001). For ICSI-spermatozoa cycles, there were no significant differences in CPR (22.6%, 29.4% and 26.1%) and LBR (16.1%, 29.4% and 19.6%) among NOA subgroups. The CPR of ICSI-spermatid cycles was 0.0%, 9.1% and 0.0% without a live birth. For ICSI-spermatocyte cycles, no clinical pregnancies occurred in any group. Conclusion: There was no significant difference in the FR of embryo-transferred TESE/ICSI cycles among NOA subgroups. The FR among all NOA subgroups was significantly lower than that of OA. Testicular histopathology in NOA did not affect successful pregnancy if spermatozoa extraction from the testis is successful and embryo transfer is possible.

Effect of the Concentration of Ammonia in Maturation Medium on the Development and Cell Numbers of Korean Native Cow Embryos (한우 난포란의 체외성숙 배지 내의 암모니아 농도가 배 발생과 세포수에 미치는 영향)

  • Park Y. S.;Park H. D.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.31-36
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    • 2005
  • The purpose of this study was an improvement of efficiency and quality in the production of Korean Native Cow embryos. We investigated effects of concentration of ammonia in in vitro maturation (IVM) medium. In addition, we examined effects of addition or exchange of IVM medium on subsequent development and the cell numbers of blastocysts. The concentrations of ammonia in IVM medium was significantly increased by the increasement of IVM duration (p<0.05). The development rates to the 2 cell-, 8 cell- and blastocyst-stage embryos with the addition of IVM medium were similar among treatment groups. The number of inner cell mass (ICM) cells and the total cell number (TCN) of blastocysts were not differ among treatment groups, whereas the trophectoderm (TE) cell number was significantly lower in the group of 4.5 h addition. The ICM/TCN ratio was significantly higher in the group of 4.5 h addition than in the group of control and 9 h addition. The development rate to the 2-cell embryo with the exchange of IVM medium was significantly higher in the group of 4.5 h exchange and 9 h exchange than in control. The development rate to the blastocyst stage was the highest in the group of 9 h exchange. The number of ICM and ICM/TCN ratio were significantly higher in the group of 9 h exchange than the other groups. The numbers of TE and TCN was similar among treatment groups.