• Clinical and Experimental Reproductive Medicine
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• 제41권2호
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• pp.41-46
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• 2014

IVM refers to the maturation of immature oocytes in culture after their recovery from small antral follicles at the stage prior to selection and dominance. IVM requires little or no FSH in vivo and has been proposed as an alternative to conventional IVF, since it reduces the primary adverse effects caused by controlled ovarian stimulation, including the ovarian hyperstimulation syndrome. Moreover, IVM is a promising option for cases for which no standard protocol is suitable, such as FSH resistance, contraindications for ovarian stimulatory drugs, and the need for urgent fertility preservation. Recently, IVM has been used in women with regular cycles and normal ovaries. However, the pregnancy rate following IVM is suboptimal compared with that of conventional IVF, indicating that further studies to optimize the protocol and the culture conditions are warranted.

• 한국수정란이식학회지
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• 제23권2호
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• pp.93-100
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• 2008

This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient macromolecule in the porcine in vitro production (IVP) technology. To choose the efficient macromolecules in the development of porcine embryos, the effects of 3 kinds of macromolecules (porcine serum; PS, porcine follicular fluid; pFF, and polyvinyl alcohol; PVA) supplemented in IVM media on the maturation, cleavage, and development rates to blastocyst of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were examined. The maturation rates of porcine oocytes in media supplemented with PS were significantly higher than those with pFF and PVA (92.4% vs. 85.4%, 77.1%; p<0.05). In the cleavage and development to blastocyst rates, supplement with PS or pFF in the IVM media was more effective than PA. However, there were no significant differences in cleavage and development to blastocyst between PS and pFF group. From the results of this study, it was demonstrated that PS was optimal macromolecule in the porcine IVM media.

• 한국가축번식학회지
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• 제18권2호
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• pp.105-111
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• 1994

The effects of human or mouse leukemia inhibitory factor(hLIF or mLIF) were examined as a means of increasing the development of in vitro matured(IVM) and in vitro fertilized (IVF) oocytes into morulae or blastocysts. Cell numbers of blastocysts were also counted using Hochest dye staining. Two-to 8-cell embryos derived from bovine IVM/IVF oocytes were cultured 5 to 6 days in CRI aa with or without mLIF or hLIF. All culture media were contained 3mg/ml bovine serum albumin. In experiment 1, the proportion of embryos developed to morulae and blastocysts in CRI aa containing 5,000U/ml mLIF(37.8%) was slightly higher than those of CRIaa containing 1,000U/ml mLIF(34.6%) and 0 U/ml mLIF(27.4%; P>0.05). In experiment 2, 0, 1,000 and 5,000U/ml of hLIF added to CR1aa media yielded 27.6%, 43.0% and 35.5% morulae and blastocysts, respectively(p>0.05). These were no significant increases in cell number among treatments(p>0.05). These results were indicating that mLIF or hLF can increase the proportion of embryos that develop into morulae and blastocysts without and increase in the cell number.

• 한국수정란이식학회지
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• 제10권3호
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• pp.229-236
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• 1995

The experiments reported here take advantage of the large number of in vitro matured and in vitro fertilized(IVM /IVF) bovine oocytes which can be produced, permitting the design of controlled experiments to establish a simple defined medium for the study of early embryo requirements. A total of 1,386 IVM /IVF oocytes were used to compare a simple defined medium(KSOM) with more complex culture conditions used successfully for culture of bovine embryos but do not permit study of specific requirements. All experiments were extensively replicated factorials. In Experiment 1, KSOM was superior to Menezo B$_2$ medium in producing morulae plus blastocysts from IVM /IVF oocytes(33 vs 20%, P<0.()5). The yield of morulae plus blastocysts with KSOM was 22% and with BRLC added was 30%. In Experiment 2, (a 2x2 factorial of KSOM with or without BRLC and 0, 1 ng /ml of platelet derived growth factor, PDGF) more morulae plus blastocysts (40%) were produced in KSOM-BRLC co-culture containing 1 ng /ml PDGF than in the control KSOM(12%). In Experiment 3, there was no dose response when 0, 1 and 5 ng /ml of PDGF were added. The results with simple defined KSOM medium are sufficiently promising to indicate that specific requirements of the embryo may be examined in future studies with KSOM as a base.

• 한국수정란이식학회지
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• 제9권1호
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• pp.85-93
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• 1994

This study was carried out to investigate the effects of gonadotropins added during maturation of porcine oocytes on the in vitro maturation(IVM), in vitro fertilization(IVF) and developmental potential of embryos. The follicular oocytes were cultured in TCM-199 medium containing different combination of gonadotropins(5$\mu$g /ml FSR or 1OIU /ml PMSG and 1O$\mu$g /ml LH or 1OIU /ml hCG), 10% FCS and 10% PFF for 36~48h in a incubator with 5% $CO_2$ in Air at 39$^{\circ}C$ and then matured oocytes were again cultured to 120h after IVF for 6~7h with heparin(100$\mu$g /m')-treated sperm. When the oocytes were matured for 42brs in the medium containing FSH+LH, FSH+hCG, PMSG+LH or PMSG+hCG, the JVF rate of each treatment was 50.0%, 52.9%, 66.7% and 70.0%, respectively. The highest CEI (cumulus cell expansion index) was obtained from PMSG+hCG-added medium and the highest polyspermic penetration resulted from FSH+LH-added medium. The cleavage of IVF oocytes derived from hormone added IVM was significantly(P<0.05) promoted by PMSG+hCG and the cleavage rate after 36-h, 42-h and 48-h maturation aws 53.0%, 56.7% and 45.6%, respectively. The highest developmental potential resulted from the oocytes derived from PMSG+LH -added IVM.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.79-79
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• 2002

본 연구에서는 in vitro에서 성숙된 난자의 핵성숙(Polar Body extrusion)에 소요되는 시간과 배반포 단계로의 발달능력 사이의 관계를 비교하여 조기에 발달능력을 가진 embryo를 선발할 수 있는 IVP 체계를 개발하고자 하였으며 in vitro maturation(IVM)에 따른 first polar body(PB) 형성, IVM과 IVF 시간이 oocyte의 발달에 미치는 영향과 생산된 배반포의 세포수를 평가하였다. IVM은 TCM199 배양액을 사용하였고 in vitro fertilization(IVF)은 Fer -TALP용액을 사용하였으며 in vitro culture(IVC)는 CRlaa 배양액을 사용하여 2일까지는 0.3% BSA를 3일 부터는 10%FBS와 bovine oviduct epithelial cell을 첨가하여 배양하였다. IVM 시간에 따른 PB의 출현율은 0hr(0%), 6hr(0%), 12hr(0%), 14hr(8.7%), 16hr(40.5%), 18hr(48.0%), 20hr(65%), 22(68%) 그리고 24hr(74.5%)을 보였으며 IVM 시간에 따른 cleavage 및 8cell 발달율 사이에는 유의적인 차이가 없었으나 배반포(BL) 및 8cell에서 배반포로 발달률은 18시간(BL 31$\pm$6, BL/8cell 82 $\pm$5%)에서 가장 높게 나타났으며 24시간(BL 17$\pm$2, BL/8cell 60$\pm$8%)과 유의적인 차이를 보였다(P<0.05). IVC 7일째 배반포의 총세포수와 trophoblast(TE) 세포수는 IVM 18시간(mean$\pm$S.E.; total: 131.1$\pm$34.0, TE: 97.6$\pm$29.6)에서 24시간(total: 112.2$\pm$17.5, TE: 80.1$\pm$15.6)보다 유의하게 많은 것으로 나왔으나(P<0.05) 7일째의 inner cell mass(ICM) 숫자(18hr 33.5$\pm$12.8 vs 24hr 32.1$\pm$12.0)와 8일째 ICM, TE 그리고 총 세포수에는 유의성 있는 차이가 없었다. IVM 18시간에서 PB 형성과 8cell 발달률 사이에 높은 상관성을 보였고 배반포 및 8cell에서 배반포 단계로 높은 발달률을 보였으며 생산된 배반포의 TE 숫자와 총 세포수가 유의하게 많은 것으로 나타났다. 따라서 IVM 18시간 실시하였을 경우 보다 많은 세포수를 가진 배반포 발달 가능성이 높은 embryo를 조기에 선발 가능할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제43권2호
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• pp.126-132
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• 2016

Objective: The purpose of this study was to identify useful clinical factors for the identification of patients with polycystic ovary syndrome (PCOS) who would benefit from in vitro maturation (IVM) treatment without exhibiting compromised pregnancy outcomes. Methods: A retrospective cohort study was performed of 186 consecutive patients with PCOS who underwent human chorionic gonadotropin-primed IVM treatment between March 2010 and March 2014. Only the first IVM cycle of each patient was included in this study. A retrospective case-control study was subsequently conducted to compare pregnancy outcomes between IVM and conventional in vitro fertilization (IVF) cycles. Results: Through logistic regression analyses, we arrived at the novel finding that serum $anti-M{\ddot{u}}llerian$ hormone (AMH) levels and the number of fertilized oocytes in IVM were independent predictive factors for live birth with unstandardized coefficients of 0.078 (95% confidence interval [CI], 1.005-1.164; p=0.037) and 0.113 (95% CI, 1.038-1.208; p=0.003), respectively. Furthermore, these two parameters were able to discriminate patients who experienced live births from non-pregnant IVM patients using cut-off levels of 8.5 ng/mL and five fertilized oocytes, respectively. A subsequent retrospective case-control study of patients with PCOS who had serum AMH levels ${\geq}8.5ng/mL$ showed that IVM had pregnancy outcomes comparable to conventional IVF, and that no cases of ovarian hyperstimulation syndrome were observed. Conclusion: Serum AMH levels are a useful factor for predicting pregnancy outcomes in PCOS patients before the beginning of an IVM cycle. IVM may be an alternative to conventional IVF for PCOS patients if the patients are properly selected according to predictive factors such as serum AMH levels.

• 한국수정란이식학회지
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• 제21권3호
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• pp.183-190
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• 2006

본 연구는 고급육을 생산한 한우의 유전자를 재활용하기 위해 기초 등록우와 혈통 등록우의 육질등급이 1 등급 이상인 암소의 난소에서 개체별로 채취한 난포란을 개체별로 각각 체외 성숙, 체외 수정 및 체외 배양하였다. 그 결과를 요약하면 다음과 같다. 기초 등록우의 육질 등급별로 회수된 난포란은 각각 1 등급 평균 28.9개, $1^+$등급 평균 28.8개, $1^{++}$ 등급 29.6개로 평균 29.0개였다. 기초 등록우의 육질 등급별 배반포배 형성율은 각각 1 등급 27.2%, $1^+$ 등급 28.7%, $1^{++}$ 등급 32.9%로 평균 8.4개의 배반포배가 생산되었으며, 배반포배 형성율은 육질등급간에 유의적 차이가 없었다. 혈통 등록우의 육질 등급별로 회수된 난포란은 각각 1 등급 평균 25.8개, $1^+$등급 평균 27.1개, $1^{++}$ 등급 27.0개로 평균 26.7개였다. 혈통 등록우의 육질 등급별 배반포배 형성율은 각각 1 등급 23.0%, $1^+$ 등급 33.7%, $1^{++}$등급 42.6%로 평균 8.6개의 배반포배가 생산되었으며, 배반포배 형성율은 육질등급간에 유의적 차이가 있었다(p<0.05).

• 한국가축번식학회지
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• 제20권3호
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• pp.279-288
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• 1996

본 연구는 EGF가 체외성숙과 수정에 의해 생산된 소 수정란의 발달과 inner cell mass (ICM)와 trophectoderm (TE) 세포수에 미치는 영향 및 공동배양시의 첨가효가를 조사하고 그와 더불어 간적면역 형광법을 이용하여 EGF-R 단백의 발현 유무를 조사하기 위해 실시하였다. 그 결과를 요약하면 다음과 같다. EGF 1, 10, 100 ng/ml의 농도로 처리되었던 4-세포기와 8-세포기 배는 대조군에 비하여 유의차는 인정되지 않았으나 양호한 배반포기 배 발달과 ICM과 TE 세포수 증가 양상을 나타내었다. 특히, 발달단계에 따른 EGF (10ng/ml) 효과를 조사하였던 바, 8-세포기 이후 배에서 대조군에 비하여 배반포기까지 유의한 배 발달을 유도하는 것을 확인할 수 있었지만 (p<0.05), ICM과 TE 세포수 증가에는 유의한 영향을 미치지 못하는 것을 알 수 있었다. 또한, 간접 면역 형광에 의한 EGF-R의 발현 유무를 조사한 결과, EGF-R는 4-세포기 이후에 발현되며 그 강도는 발달단계가 진행되면서 다양하게 나타난다는 것을 알 수 있었다. 한편, 수정란과 난구세포 공동배양 군은 EGF의 첨가 유무에 상관없이 대조군에 비하여 유의한 배 발달과 총세포수의 증가를 나타내며, 공동배양군에 대한 EGF 첨가는 수정란과 난구세포와의 공동배양 효과를 증진시키는 것으로 나타났다. 따라서, EGF는 착상전 소 수정란의 4-세포기 이후에 발현디는 EGF-R에 반응하여 배 발달을 유기하고, 공동배양시의 배 발달에 유용한 물질형성을 촉진시키지만, 배반포기 배의 ICM과 TE 세포수 증가에는 유의한 영향을 나타내지 못한다는 것을 알 수 있었다.

• 한국가축번식학회지
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• 제20권2호
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• pp.191-196
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• 1996

The objective of this study were to investigate the effects of culture media and additives on the development of bovine in vitro matured(IVM) and in vitro fertilized(IVF) oocytes. In experiment 1, bovine oocytes were cultured in droplets of TC 199 supplemented with 10% fetal bovine serum(FBS) with or without hormones (5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml E2). Cleavage rates of embryos cultured for 40~44hrs after IVF were higher when embryos were cultured in TC 199 supplemented hormones (68.1%, 921/35) than without hormones (52.7%, 77/146), but the percentages of development beyond morulae stage were not difference (20.7%, 19.4%). In experiment 2, the effects of various media such as TC 199, synthetic oviduct fluid(SOF), CR1aa with different energy source (fatal bovine serum, FBS; bovine serum albumin, BSA) on developmental capacity of IVM/IVF bovine embryos were investigated. The developmental rates into morulae and blastocysts were 27.1, 10.7, 6.3 and 0%, respecitvely, in CR1aa plus 3mg/ml BSA, SOF plus 10% FBS, TC 199 plus 10% FBS, SOF plus 3mg/ml BSA. In experiment 3, the comparisons of bovine embryos developed to morulae and blastocysts in different culture media (TC 199, SOF, CR1aa, Menezo's B2) were investigated. The developmental capacity beyond morulae stage were 32.9, 26.6, 11.1 and 7.1%, respectively, in Menezo's B2 plus BSA, CR1aa plus BSA, SOF plus BSA, TC 199 plus FBS medium. The cell numbers of the blastocyst were not different in different cultrue media. In experiment 4, bovine embryos were co-cultured with vobine oviduct epithelial cells(BOEC) in TC 199 plus FBS, SOF plus BSA, CR1aa plus BSA, Menezo's B2 plus BSA. The morula and blastocyst rates were 44.7, 32.9, 26.0 and 23.3%, respectively, in CR1aa TC 199, SOF, and Menezo's B2 medium. The cell numbers of the blastocyst were similar to those of blastocyst developed in different culture media.