• Title/Summary/Keyword: 체외 수정

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무혈청배양액으로 생산한 한우 체외수정동결란의 이식 결과

  • 정연길;임여정;임광택;설현석;송해범
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.281-281
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    • 2004
  • 최근 체외수정, 핵이식, 체세포 복제동물의 생산 등으로 수정란의 동결 보존기술의 개발이 매우 중요한 시기에 왔다. 본 실험에서는 한우의 체리수정란을 완전 무혈청 배양액으로 배양하여 수정란의 배발생율과 동결융해 한 후 생존성 및 수정란이식 한 후 수태율을 조사하였다. (중략)

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Effects of Kinds and Concentrations of Cryoprotectants, Trehalose, Sucrose-Addition in Cryoprotectants on the Survival Rates of Vitrification-Thawed Porcine IVM/IVF Embryos (돼지 난포란의 내동제의 종류와 농도 및 Trehalose, Sucrose등의 첨가가 Vitrification 동결 융해 후 생존율 및 수정율에 미치는 영향)

  • Lim J. G.;Quan J. H.;Lee K. S.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.123-128
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    • 2005
  • This study examines the effects of kinds and concentrations of cryoprotectants on the survival rate of vitrification-thawed porcine oocytes, together with the effects on survival, in vitro fertilization and development of immature oocytes. 1. The developmental rate of oocytes to MII and diploid stage when the vitrification-thawed of recovered immature oocytes cultured for 0, 15, 30 and 40h were cultured for 0, 15, 30 and 40h were $56.7\%,\;53.3\%,\;63.3\%,\;65.0\%\;and\;23.3\%,\;18.3\%,\;10.0\%,\; 3.3\%$, respectively. The in vitro development to MII stage were lower than the control group $(78.2\%)$, but higher fo. diploid stage $(5.5\%)$. 2. When the vitrification of immature oocytes after being culture for 0, 15, 30 and 40 hours, the survival rate were $34.0\%,\;26.0\%,\;18.0\%\;and\;10.0\%$ respectively. This result was lower than that of the control group $(60.0\%)$. 3. When the fertilization of the vitrified immature oocytes after being culture for 0, 15, 30 and 40 hours, the in vitro fertilization rate were $60.0\%,\;54.0\%,\;48.0\%,\;38.0\%$, and developmental rates were $26.0\%,\;18.0\%,\;8.0\%,\;4.0\%$, respectively. This results were lower than the control group $(78.0\%\;and\;38.0\%)$. 4. When the fertilization of the immature oocytes after being culture for $0\~15$ hours vitrified with EDS and ETS, the fertilization and developmental rates were $50.0\%,\; 22.0\%$ and $46.0\%,\;18.0\%$, respectively. This results were lower than the control group $(74.0\%\;and\;38.0\%)$.

Effect of Oxygen Concentrations with Catalase on In Vitro Maturation of Porcine Follicular Oocytes and In Vitro Development of Porcine IVM/IVF Embryos (산소조건 및 Catalase가 돼지난포란의 체외성숙과 배발달에 미치는 영향)

  • 한만희;이경본;천행수;박병권;이경광;이규승;서길웅
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.115-123
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    • 2003
  • The present study was carried out to examine the effect of catalase (CAT) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with CAT on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows - 1 . The rates of nuclear maturation, penetrated oocytes, pronucleus formation rates, polyspermic oocytes and mean numbers of the penetrated sperm were significantly lower in oocytes matured with 100, 500 and 1,000 units/ml CAT than those of central groups (P>0.05). 2. The rates of blastocyst formation and total cell numbers of blastocyst at day 7 after in vitro fertilization were significantly lower in CAT treatment groups than those of the central groups (P>0.05). 3. There were not significant difference in the blastocyst development and total cell numbers of blastocyst on in vitro culture of NCSU-23 media with 0, 100, 500 and 1000 units/ml CAT under the 5% and 20% $O_2$ concentrations. These results suggested that the addition of CAT was not helpful for porcine oocyte maturation and further development, also the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under the 5% and 20% $O_2$ concentrations.

Morphologic Parameters and in vitro Maturational Competence of Human Immature Oocyte Obtained from Stimulated IVF Cycle (미성숙난자의 형태학적 지표와 체외성숙능과의 관계)

  • An, So-Jung;Jee, Byung-Chul;Moon, Jeong-Hee;Lee, Jung-Ryeol;Suh, Chang-Suk;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.285-291
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    • 2008
  • Objective: This study was performed to investigate whether cumulus morphology and oocyte diameter influence on in vitro maturation (IVM) of human germinal vesicle (GV) stage oocytes obtained from stimulated in vitro fertilization (IVF) cycles. Methods: Forty-one GV stage oocytes were obtained from 21 patients who received ovarian hyperstimulation and IVF. According to cumulus morphology before denudation, GV oocytes were classified into oocytes with dispersed cumulus cells (CCs) or compacted CCs. The diameters of denuded oocytes, both including and excluding the zona pellucida, were measured. All oocytes were cultured in commercial IVM medium. Maturation was defined as extrusion of the first polar body and the matured oocytes were inseminated by ICSI method. Results: Overall maturation and fertilization rate were 56.1% and 73.9%. Matured oocytes had significantly higher proportion of oocytes with dispersed CCs compared to oocytes failed to mature (91.3% vs. 55.6%, p=0.023). There were no significant differences of oocytes outer ($155.7{\mu}m$ vs. $152.4{\mu}m$, NS), inner ($114.3{\mu}m$ vs. $113.4{\mu}m$, NS) diameters and zona thicknesses ($41.3{\mu}m$ vs. $39.1{\mu}m$, NS) between matured and not-matured oocytes. In-vitro maturation rate of oocytes with dispersed CCs was significantly higher than which of oocytes with compacted CCs (67.7% vs. 20.0%, p=0.044). Oocyte diameters (outer and inner) and thicknesses were not related with maturational competence. Conclusion: Our results suggest that in vitro maturational competence of GV stage oocytes obtained from stimulated IVF cycles is closely associated with the cumulus morphology but not oocyte diameter.

The Clinical Outcome of Twin Pregnancies after IVF (체외수정 후 출산한 쌍태아의 임상적 고잘)

  • Han, Myoung-Seok;Park, Eun-Ku-Seul
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.3
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    • pp.173-178
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    • 2007
  • Objective: To compare the outcome of twin pregnancies after in-vitro fertilization (IVF) with that of spontaneously conceived twins. Methods: We analyzed 146 twins retrospectively, who had been delivered between January 2000 and July 2005. After reviewing their obstetric medical records, 72 twins conceived with IVF (IVF group) and 51 spontaneously conceived twins (spontaneous group), whose gestational age was over 24 weeks, were recruited for this study. Results: There were no significant statistical differences of demographic features such as maternal age, gestational age and fetal weight between the two groups. However, the risk of pregnancy-induced hypertension (PIH) and gestational diabetes (G-DM) had significantly increased in the group of IVF (OR 2.59; 95% CI 1.01$\sim$6.66). The risk of fetal weight discordancy rate (>20%) and fetal-sex homology rate in IVF group were decreased (OR 0.37; 95% CI 0.14$\sim$0.96, OR 0.45; 95% CI 0.21$\sim$0.99). Conclusion: In this comparative study, maternal PIH and G-DM risks of IVF group are higher than that of spontaneous group. But, fetal-weight discordancy risk and fetal-sex homology rate were lower in IVF group.

In Vitro Fertilization and Development of Bovine Immature Oocytes by Ultra- Rapid Cooling (미성숙 소난자의 초급속 동결ㆍ융해 후 수정 및 체외 배발달)

  • Ryu, C. S.;K. S. Kil;Kim, N-H;D. H. Ko;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.23 no.3
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    • pp.221-227
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    • 1999
  • Successful cryopreservation of bovine immature oocytes can increase availably of oocytes for the in vitro fertilization or nuclear transfer. However, it was not reported successful development to the blastocyst stage following in vitro fertilization of cryopreserved bovine immature oocytes. The objective of this study was to determine the incidence of survival, meiotic maturation, fertilization and in vitro development of cryopreserved bovine immature by ultra rapid cooling methods. The oocytes were adversely affected by brief exposure to EFS40 solution in electron microscope grids and plunged directly into liquid nitrogen. After such ultra-rapid cooled immature oocytes were warmed, 78% of oocytes were matured to the metaphase II stage, 50% of oocytes were fertilized after insemination, and 5% of oocytes were developed to the blastocyst stage. Different sodium concentration of sodium ion in the freezing medium did not affect survival, maturation, fertilization and in vitro development of cryopreserved oocytes. These results suggested that immature bovine oocytes can be cryopreserved by ultra-rapid cooling methods.

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Effects of Protein Sources and Co-culture on In Vitro Culture of IVF-derived Porcine Embryos (단백질 공급원 및 체세포와의 공배양이 돼지 체외수정란의 체외발달에 미치는 영향)

  • 한선경;구덕본;이규승;황윤식;김정익;이경광;한용만
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.289-297
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    • 2000
  • This study was conducted to investigate whether various protein sources and co-culture affect in vitro development of porcine zygotes derived from In vitro maturation/fertilization (IVM/IVF). These results obtained in these experiments are summarized as follows 1. When porcine oocytes matured and fertilized In vitro were cultured in NCSU 23 medium supplemented with various BSA concentrations (0.4, 0.8 and 3.2%), In vitro developmental rates of porcine zygotes to blastocyst stage were 22.9, 18.4 and 14.6%, respectively. High concentration of BSA (3.2%) showed a smaller nuclei number (36.1$\pm$11.8) of blastocysts than 0.4 and 0.8% BSA groups (53.2$\pm$27.4 and 61.2$\pm$22.5, respectively) (P<0.05). This result indicates that high concentration of BSA is detrimental on preimplantation development of IVF-derived porcine embryos. 2. No differences were detected in the developmental rate and mean nuclei number of porcine embryos between 10 and 20% FBS concentrations in culture medium. 3. IVF-derived porcine embryos co-cultured with mouse or porcine embryonic fibroblast cells showed a lower development to the blastocyst stage than those without co-culture system. Consequently, the present study suggests that high concentration of BSA as a protein source in culture medium suppresses development potential of porcine embryos produced In vitro. In addition, co-culture with somatic cells is not effective on in vitro development of IVF-derived porcine embryos to blastocyst stage.

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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.