• Proceedings of KOSOMES biannual meeting
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• 2009.06a
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• pp.163-163
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• 2009

• The Journal of Korean Obstetrics and Gynecology
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• v.28 no.2
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• pp.109-119
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• 2015

Objectives : This study is to report the effect of herbal medicine and acupuncture on a clinical pregnancy. Methods : From february 2014 to july 2014, a prospective analysis study was performed in 20 patients after taking her medicine and acupuncture treatment. Results : After treatment, two patients naturally became pregnance and one patient became pregnance after in vitro fertilization-embryo transfer (IVF-ET). Conclusions : This study suggests that herbal medicine and acupuncture treatment are useful and shows possibility to increasing pregnancy rates.

• Journal of Bio-Environment Control
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• v.18 no.4
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• pp.454-459
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• 2009

The secondary growth of garlic depreciates the quality of a bulb and, in the worse cases, makes it difficult to be used as a seed bulb due to many insertions of small cloves in the seed bulb. Therefore, the effect of soil temperature, seedtime, and fertilization rate on the secondary growth of the big bulbils of Namdo garlic has been examined. When we analyzed into the growth characteristics of big bulbils of Namdo garlic under different fertilizer application levels (50% increased fertilization, experimental fertilization and 50% decreased fertilization), the plant height, numbers of leaf, sheathe diameter, leaf length, and leaf width were the best in 50% increase of the test rate of fertilizer. The occurrence rates of secondary growth were 34.2% in 50% decrease of the test rate of fertilizer, 44.3% in the test rate, and 54.1% in 50% increase of the test rate of fertilizer. In other words, the occurrence rate of secondary growth increased by increasing the fertilization rate. While the average harvested bulb weight were 34.1g in 50% decrease of the test rate of fertilizer, 35.1g in the test rate of fertilizer, and 33.9g in 50% increase of the test rate of fertilizer, there were no changes in the number of cloves under different fertilization rates. As the correlation diagram between the soil temperature and occurrence of the secondary growth showed very high relation with 0.892~0.997, the secondary growth in cultivation of Namdo garlic big bulbils had considerably close correlation with the soil temperature. As the earlier the seedtime was, the growth of the above-groundparts including the plant height, numbers of leaf, and sheath diameter were the better. The averages of bulb weight were 36g in the mid September sowed seed bulb, 29.6g in the late September sowed seed bulb, and 27.9g in the early October sowed seed bulb. Overall, our results showed that the bulb size is dependent on the seedtime and the seedtime has no particular effects on the secondary growth.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.1
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• pp.51-59
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• 1996

Controlled Ovarian hyperstimulation(COH) is generally used to obtain synchronous high quality oocytes in in vitro fertilization-embryo transfer(IVF-ET). Many investigators have studied the relationship between serum hormone levels and outcomes of IVF-ET because there is no accurate estimation method of oocyte quality. Early premature luteinization of follicles before oocyte retrieval is the most troublesome problem in COH for IVF-ET. Gonadotropin-releasing hormone agonists(GnRH-a) are used as adjuncts with gonadotropins for COH in patients undergoing in IVF. The possible benefits of GnRH-a pretreatment include improving oocyte quality, allowing a more synchronous cohort of follicles to be recruited, and preventing premature lueinization hormone surges. In COH of IVF cycles, we investigated whether an elevated progesterone(P4) level on the day of human chorionic gonadotropin(hCG) administration indicates premature luteinization and is associated with a lower fertilization rate. Many investigators have studied that the lower fertilization rates seen in patients with elevated P4 levels might result from an adverse effect of P4 on the oocytes. We hypothesizes that serum P4 levels around the day of hCG may be helpful prediction of out come in IVF-ET cycles. Success rates after COH of IVF-ET cycles are dependent upon many variable factors. Follicular factors including the number of follicles, follicular diameters and especially serum estradiol(E2) levels as an indirect measurement of follicular function and guality have been thought to influence the outcomes of IVF-ET. To assess whether serum P4 and E2 levels affect the fertilization and pregnancy rate, we reviewed the stimulation cycles of 113 patients (119 cycles) undergoing IVF-ET with short protocol with GnRH-a, from March 1993 to August 1994 retrospectively. The serum P4 and E2 levels were compared on the day of hCG in the pregnant group, 45 patients(47 cycles) and in the non-pregnant group, 68 patients (72 cycles) respectively. The serum E2 level in non-pregnant group was $1367{\pm}875.8$ pg/ml which was significantly lower than that of pregnant group, $1643{\pm}987.9$ pg/ml( p< 0.01 ). And the serum P4 level in non-pregnant group was $2.1{\pm}1.4$ ng/ml which was significantly higher than that of pregnant group, $1.0{\pm}0.7$ ng/ml( p< 0.001 ). The fertilization rate was $61.3{\pm}21.3%$ in pregnant group which was higher than that of non-pregnant group, $41.1{\pm}20.2%$ (p< 0.01). We suggest that the serum levels of P4 and E2 on the day of hCG administration are additional parameters that predict the outcomes of IVF-ET cycles.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.207-212
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• 2009

The aim of present experiment was to examine commercial synthetic extender(AndroMed) for semen cryopreservation of Korean Black Bull. Semen was collected from a Korean Black Bull using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by AndroMed. The pellect was diluted to final sperm concentration of $5{\times}10^5/ml$ by doubling in every 10 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 1 hr at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 minutes and above 10 cm for 10 min. And then the frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Hanwoo semen was used as KPN (Korea Proven Bull Number) in this experiment. The survival rates was significantly higher in fresh semen than frozen semen ($80{\pm}14%\;and\;43{\pm}11%$). However, the motility rates was similar (80.7% and 66.4%). The survival and motility rates were higher in 5cm, 10 min treatment group than the other two groups in straw-located height and duration above $LN_2$ ($50{\pm}14%$ and 70.7% vs, 33.18% and $65{\pm}7%$ vs, 30.14% and 65.7%, respectively). The development rates to cleavage was higher in Black Cow than Hanwoo semen (62.2%, 64.4%), However, The development rates to blastocyst was higher in Hanwoo than Black cow semen (25.9%, 23.0%). In conclusion. The present results that acceptable fertilization and cryopreservation could be obtained by in vitro fertilization with frozen-thawed semen using a synthetic semen extender (AndroMed).

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.173-182
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• 2001

The present study was carried out to examine the effect of superoxide dismutase (SOD) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with SOD on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows : 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes and mean numbers of the penetrated sperm were not different in the NCSU-23 maturation media with 0, 100, 500 and 1,000 units/ml SOD. However. the pronucleus formation rates were significantly lower in oocytes matured with addition groups than those of no addition groups of SOD (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 addition groups than those of the no addition groups of SOD (P>0.05). 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 culture medium with 100 units/ml SOD than in those cultured with 0, 500 and 1,000 units/ml SOD under the 5% and 20% $O_2$concentrations. However, no differences was found in the total cell numbers of blastocyst among the treatments. In conclusion, these results suggested that the addition of SOD was not adequate 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 different in the NCSU-23 culture medium under the 5% and 20% $O_2$concentrations.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.113-120
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• 1993

To provide the optimal culture conditions for the developm,ent of in vit개 produced embryos, we have been investigated various culture media as well as co-cultrue systems using porcine cumulus cells or mouse fetal fibroblast cells. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles(3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated(39$^{\circ}C$, 5% CO2 in air) in various maturation media for 42 hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were rpepared for fertilizing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${\mu}\ell$ fo capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three different media, m-KRB, BECM and TCM-HEPES were 0~1.0%, showing extremely lower rates. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. The rates of embryos developed to 2-, 4-, 8-, 16-, 32-cell and morula or blastocyst stage in co-culture with porcine cumulus cells and mouse fetal fibroblast cells were 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% and 20.4~21.0%, respectively. These development rates upto morula or blastocyst stages were significantly higher than those of the embryos cultured in the basic culture medium(P<0.01). These findings suggest that co-culture of in vitro fertilized eggs with porcine cumulus cells or mouse fetal fibroblast cells enhance the development of fertilized eggs to morula or blastocyst stage in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.166-172
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• 2019

Objective: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts. Methods: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined. Results: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p< 0.05). Conclusion: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.189-203
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• 1999

To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.113-113
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• 2003

This study was conducted to investigate the effect of vitrification solution(VS) on in vitro developmental competence of immature porcine oocytes. The immature porcine oocytes were exposed to the following vitrification solution, at RT. 1) EFS sol. : 20% ethylene glycol (EG) 3 min, 40% EG + 18%(w/v) Ficoll(MV70, 000) + 0.3 M sucrose 30 sec, 2) GE sol. : 10% glycerol 5 min, 10% G + 20% EG 5 min, 25% G +25% EG 30 sec, 3) EG sol : 1.5M EG 2.5 min, 5.5 M EG + 1.0 M sucrose 30 sec. Oocytes were immediately transferred into 1.0 M, 0.5 M, 0.25 M, 0125 M, 0 M sucrose solution for 2.5 min each at RT. After removal of VS, immature oocytes were matured in vitro and subsequently all oocytes were subjected to IVF followed in vitro culture for 7 days. Maturation rates of oocytes were 38.8%, 44.5%, 22.4% and 57.6%, in EFS, EG, GE and Control, respectively, maturation rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.01). Fertilization rates of oocytes in Control was significantly higher than other treated groups(P<0.05), but no difference were observed among treated groups. Polyspermic rates were no significant difference among four groups. Cleavage rates of oocytes were 21.9%, 47.1%, 19.0% and 65.9%, in EFS, EG, GE and Control, respectively, cleavage rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.05), but blastocyst formation rates were no significant difference among four groups. These results suggested that the use of EG solution could be a great challenge for reaching a successful vitrification of immature porcine oocytes.