• 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.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.265-273
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• 2004

This study was carried out to examine the effects of fertilization media and sperm concentration on in vitro fertilization (IVF) and development (IVD) of porcine oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF). After the fertilization by experimental scheme, putative embryos were developed in vitro in NCSU-23. The results are as follows. When the oocytes were fertilized in vitro in modified TBM or modified TLP-PVA by 1 ${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were not significantly different between two media. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were not different between two media, either. When the oocytes were fertilized in vitro in mTBM by 5${\times}$10$^4$, 1${\times}$10$^{5}$ or 5${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were significantly (P<0.05 or P<0.01) increased as sperm concentration was elevated. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were significantly boosted (P<0.01) as sperm concentration at fertilization was elevated from 5${\times}$10$^4$ to 1${\times}$10$^{5}$ sperm/$m\ell$, but were not different between 1${\times}$10$^{5}$ and 5${\times}$10$^{5}$ sperm/$m\ell$.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.249-254
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• 2009

Objective: The aim of this study was to compare the fertilization and cleavage rates of human in vitro matured oocytes after fertilized by conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Methods: A total of 135 GV stage oocytes were obtained from 59 women who received ovarian stimulation and IVF during Jan 2007 to Oct 2008. Ovarian hyperstimulation was performed using hMG or recombinant FSH with GnRH antagonist and then ovulation triggered by recombinant hCG. The immature oocytes obtained from stimulation cycles were cultured in IVM medium up to 48 hrs; commercial medium supplemented with rFSH 75 mIU/mL, rhCG 0.5 IU/mL and rEGF 10 ng/mL. The in vitro matured oocytes were fertilized by conventional IVF (41 GV oocytes) or ICSI method (94 GV oocytes). Results: Maturation rate were 51.2% and 59.6% in conventional IVF group and ICSI group, respectively. There was no significant difference in fertilization rates between two groups; 71.4% and 80.4%, respectively. The cleavage rate was also similar in two groups. Conclusion: The presented data suggest that conventional IVF has comparable fertilization and cleavage potential compared with ICSI as the insemination method of immature human oocytes obtained from stimulated cycle.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.72-72
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• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.97-97
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• 2004

• Journal of Embryo Transfer
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• v.19 no.1
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• pp.19-25
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• 2004

The objective of this study was to optimize the selection of sperm, optimal culture system of in vitro derived porcine embryos. The results obtained were summarized as follows: 1. When oocytes were inseminated with liquid sperm and frozen-thaw sperm, the cleavaged rate of liquid sperm (46.2%) was higher than that of frozen-thaw sperm (39.7%), however there were not show significant different each other. The blastocyst rates of liquid sperm (15.8%) was significantly higher than that of frozen-thawed sperm (9.3%)(P< 0.05). 2. When oocytes were inseminated with epididymal sperm after 1, 2 and 3 day storage, the cleavaged rate of epididymal sperm after 1, 2 and 3 day storage was 60.5, 61.0 and 56.8% respectively. The morulae (17.4, 19.9 or 17.3%) and blastocyst (8.7, 15.4, 11.3%) rate of epididymal sperm after 1, 2 and 3 day storage was no significantly respectively(P< 0.05). 3. In vitro developed to cleavaged rate of G1.3/G2.3 media used for culture was significantly(P< 0.05) higher as 62.1% compared with the results using the media NCSU23(52.8), however in vitro developed to blastocyst rate of NCSU23(11.6%) media was significantly(P< 0.05) higher than that'of G1.3/G2.3(4.7%). 4. When the fertilized oocytes were cultured with NCSU23 in addition to 1 mM glutathione(GSH), the cleavaged rate of treated groups of GSH(62.3%) was significantly higher than that of control(53.5%) respectively(P< 0.05). And in vitro developed to blastocyst rates of treated groups of GSH(15.6%) was higher than that of control(12.6%) however, there was no significant difference(P< 0.05).

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.311-317
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• 2000

This study was carried out to compare the temperature and time of heat shock, and the effect of heat shock on development of embryos after in vitro maturation and fertilization in bovine oocytes. The results obtained were as follows. 1. The optimum temperature and time of heat shock were 41$^{\circ}C$ and 30sec on in vitro development of embryos from 4~8 cell to blastocyst. 2. The rates of cleavage on zygotes produced on in vitro were significantly increased by heat shock after IVM than before IVM(P<0.05). 3. When the oocytes were treated heat shock after IVM and 5 days cultured, developmental rates to blastocyst were increased than other experimental treatments.

• Korean Journal of Animal Reproduction
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• v.22 no.2
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• pp.163-169
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• 1998

본 실험은 체외 생산된 소 배반포기배에 대한 laser drilling 처리가 배의 부화율에 미치는 영향을 조사하고자 실시하였다. 그 결과는 다음과 같다. 소 수정란의 체외 발달율을 조사하였던 바, 82.3%의 난할율( 2-세포기)과 체외수정 후 배양 7일째에 32.6%의 배반포 발달율을 나타내었다. 이렇게 생산된 배반포기배에 laser drilling 효과를 조사하였던 바, 처리 후 24시간째의 부화진행율(90.0%)은 대조군(44.4%)보다 유의하게 높게 나타났다(p<0.0001). 또한, 처리 후 48시간째의 부화율(68.0%)도 대조군(33.3%)보다 유의하게 높게 나타났다. 이러한 결과는 laser drilling이 체외 생산된 소 배반포기배의 부화진행율과 부화율을 유의하게 증가시킬 수 있다는 것을 알 수 있었다(p<0.001).

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.133-139
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• 1999

This study was carried out to investigate in vitro/in vivo development of vitrified-thawed immature mouse oocytes. Immature mouse oocytes were vitrified with EFS40 (40% ethylene glycol, 18% ficoll and 0.5 M sucrose). Thawed oocytes were matured for 16 hr in vitro. Matured oocytes with the first polar body were fertilized with the concentration of 1~2$\times$10$^{6}$ $m\ell$ of epididymal sperm. After fertilization, cleavage ($\geq$ 2-cell) and in vitro/in vivo development rates were examined. $\pi$ Ie results were summarized as follows: in vitro maturation rate of immature mouse oocytes in vitrified-thawed group was similar to that in exposed group (67.5%) and control (66.3%), but cleavage rate of vitrified-thawed oocytes (64.9 %) and blastocyst formation rate (59.0%) were significantly different compared to those of exposed group (83.7 and 74.7%) and control (90.7 and 83.7%) (p＜0.05). However, when the blastocysts derived from immature mouse oocytes vitrified-thawed were transferred to pseudopregnant mouse, total implantation (31.3%) was slightly lower than that in control (40.8%), but live fetus formation rate (66.7%)was slightly higher than that in control (58.1%), there was not significantly different. Therefore, when the blastocyts produced in vitro were transferred into recipients, although the development in vitro of oocytes vitrified-thawed was decreased, live fetus formation rate was similar to that of control group. The present results indicate that immature mouse oocytes can be frozen successfully by vitrification with EFS40.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.58-58
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• 2003

포유동물 체외 수정란의 체외 발육율을 향상시키기 위해서는 배양액내에 생성되는 free radical을 제거하는 것이 한가지 요인으로 지적되고 있다. 체외 배양액내에 생성되는 free radical을 제거하기 위하여 여러 종류의 항산화제 첨가 배양이 많이 이용되어 체외발육성적을 향상시키고 있다. 포유동물의 번식에 있어서 Melatonin은 광주기 조절 작용에 기여하는 호르몬으로서 혈중 Melatonin의 농도는 번식 작용에 중요한 역할을 하며, 또한 수정란의 세포분열과 세포주기에 영향을 미치며 배양액내의 free radical을 제거하여 수정과 초기 수정란 발달에 영향을 미쳐 항산화 기능이 있는 것으로 보고되고 있다.