• Development and Reproduction
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• v.20 no.2
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• pp.127-133
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• 2016

Purine metabolism is known factor for nuclear maturation of oocytes through both follicle cells and oocyte itself. However, it is largely unknown the roles of purine metabolism in the oocyte competence for fertilization and early development. In this study, the effects of adenosine in oocyte competence for development were examined using adenosine and its synthetic inhibitors. Adenosine treatment from GV intact stage for 18 hr (fGV) caused of decrease the fertilization rate but of increase the cleavage rate compared from the other stage treatment groups. Hadacidin did not effect on fertilization rate but increased cleavage rate without stage specificity. Adenosine did not block the effects of hadacidin with the exception of fGV group. Inhibition of purine synthetic pathways the fertilization rate was decreased in the fGV and fGVB groups but increased in the fMII group. Exogenous adenosine caused of decrease fertilization rate in the fGVB group but increase in the fMII group. Cleavage rate was dramatically increased in the adenosine treatment with synthetic inhibitors. It means that the metabolism of purine has stage specific effects on fertilization and cleavage. Exogenous adenosine had only can improve oocyte developmental competence when it treated at GV intact stage. On the other hand, endogenous synthesis in all maturation stage cause of increase the cleavage rate without effects on fertilization. These data suggest that adenosine at GV stage as a paracrine fashion and inhibitions of endogenous adenosine in all stage improve oocyte developmental competence.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.225-231
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• 1996

This study was conducted to investigate the effects of preincubation time, concentration and exposure time of sperm on in vitro fertilization of porcine follicular oocytes rnatured in in vitro. The results obtained are as follows ; 1. Effect of preincuhation time for porcine sperm capacitation on in vitro fertilization in medium with heparin was investigated. Normal fertilization rate was highest in 15 min(26.4%). However, there were no significant differences among preincuhation times of 5~90 min, 2. Normal fertilization rates of sperm concentrations were 17.0~26.5%, and normal fertilization rate from l$\times$ l05cell /ml concentration was also higher than those of other sperm concentration. 3. Normal fertilization rates of sperm exposure time of 4, 8, 12, 16 and 20 hours were 6.1, 20.8, 27.8, 25.0 and 26.7%, respectively. Normal fertilization rate from sperm exposure time of 12 hours was also higher than that of other sperm exposure times.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.1
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• pp.50-56
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• 2015

This research was performed to test the hypothesis that the optimal fertilization rate for lettuce is various with soil moisture conditions. The experiment was conducted under a rainfall-intercepted facility in Suwon, South Korea from 2002 to 2003. Soil was irrigated at 30, 50, or 80 kPa of soil moisture tension at 15 cm soil depth in 2002 spring and fall and 20, 30, 50, or 80 kPa in 2003 spring. Fertilization was performed with four levels in spring for both years: none, 0.5, 1.0, and 1.5 times of the recommended N, P, and K fertilization rate. The irrigation amount increased with decreased irrigation starting point as soil moisture tension. The maximum yield was found at the lowest soil moisture tension in spring while irrigation at 50 kPa resulted in the greatest yield in fall. The yield responses of lettuce to fertilization rates were various with soil moisture condition. In spring, maximum yield was found at 1.0 or 1.5 times of the recommended fertilization rate at 20, 30, and 50 kPa irrigation while 0.5 or 1.0 times of fertilization rate resulted in the maximum yield in fall. Especially for 80 kPa irrigation in 2003 spring, yield was decreased by fertilization. It suggested that the optimum fertilization rate for lettuce is affected by soil moisture condition and that lower fertilization rate should be suggested when soil is managed in drier condition.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.65-72
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• 2001

Objectives: This study was to investigate the fertilization rate after intracytoplasmic sperm injection (ICSI) or partial zona dissection (PZD) of human and hamster sperm into hamster oocyte in in vitro fertilization (IVF). In addition, the possibility of clinical application was evaluated by the comparison of usefulness and difference of these method. Materials and Methods: Hamster immature oocytes were obtained from oviducts superovulated by PMSG and hCG, and hamster sperms were obtained from epididymis. The freezed human sperms were thawed before use. Fertilization were confirmed by two pronuclei, one pronucleus, swollen sperm head or/and two polar bodies at $7{\sim}8$ hour after ICSI or PZD. Results: The fertilization rates after ICSI and PZD of human sperm to hamster oocyte were 3.6%, 64.2%,73.6%, and 55.6% for negative control, positive control, ICSI, and PZD respectively, suggesting that ICSI only showed improved fertilization rate (p<0.01). The fertilization rates after ICSI and PZD of hamster sperm to hamster oocyte were 11.1%, 51.2%, 39.6%, and 72.7% for negative control, positive control, ICSI, and PZD respectively, suggesting that PZD only showed improved fertilization rate (p<0.01). PZD showed significantly higher fertilization rate than ICSI (p<0.05). Conclusions: As for the fertilization rate by ICSI and PZD using hamster oocyte in IVF, ICSI technique was considered to be more useful for human sperm and PZD technique for hamster sperm. Therefore, ICSI technique was considered more appropriate for experimental application using human sperm and hamster oocyte.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.5
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• pp.332-339
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• 2014

This research was performed to test the hypothesis that the optimal fertilization rate for red pepper is changed by soil moisture condition. The experiment was conducted in rainfall-intercepted fields in Suwon, South Korea from 2002 to 2003. Soil was irrigated at 30, 50, or 80 kPa of soil moisture tension at 20 cm soil depth in 2002 and 30, 50, 100, or 150 kPa in 2003. For both years, fertilization was performed with four levels: none, 0.5, 1, and 1.5 times of the recommended N, P, and K fertilization rate. The irrigation amount was the greatest at 30 kPa irrigation while the water use efficiency increased with decrease of irrigation amount. The Irrigation amount was 508 mm at 30 kPa irrigation and ranged from 355 mm to 435 mm at 50 kPa irrigation. The maximum yield was found at 30 kPa irrigation and 1.5 times of the recommend fertilization rate in 2002 and 2003. The yield index of red pepper increased linearly with the fertilization rate at 30 kPa which implied that excess irrigation induced nutrient leaching and reduced nutrient availability. The maximum yield in 50 kPa and 80 kPa was found at the recommend fertilization rate while the yield decreased by fertilization at 100 kPa and 150 kPa irrigation. It implies that reduction of fertilization is the feasible practice to mitigate drought stress in fields without stable irrigation resources.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.2
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• pp.123-127
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• 1995

In vitro embryo production (IVP) is affected by various factors during in vitro maturation, fertilization, and development. In this experiment, the effect of ovary type, quality of follicular oocyte, medium used for fertilization, presence of hormone in medium, sperm concentration on in vitro maturation and fertilization were examined for effective IVP. In vitro maturation was carried out using TCM-199 supplemented with 15% FCS and hormones in 5% $CO_2$ incubator for 24h. In vitro fertilization was performed with frozen-thawed sperm in modified mTALP medium containing 0.3% BSA, $10{\mu}g/ml$ heparin, and 5mM/ml caffeine for 24h. The fertilized embryos were co-cultured on monolayer of cumulus cells in TCM-199. When oocytes were collected from functionally active and inactive ovaries, maturation rate was 76.9 and 7.7%, respectively. When oocytes were classified morphologically to good and poor grades, maturation rate was 75 and 58.8%, respectively. FSH + LH + $E_2$ (86.4%) showed higher maturation rate than control (53.0%) and FSH (73%). The fertilization rate was 28.2, 100 and 91.7% in $1.6{\times}10^5$, $5.0{\times}10^5$ and $10.0{\times}10^5$ sperm concentration per ml. When oocytes were fertilized in mTALP and BO media, fertilization and cleavage rates of oocytes in mTALP were higher (84.3 and 56.9%) than those (67.4 and 23.3%) in BO medium. In this experiment, in vitro maturation, fertilization and development of oocytes were affected by type of ovary, grade of oocyte, hormones, sperm concentration and fertilization medium.

• Korean Journal of Animal Reproduction
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• v.15 no.2
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• pp.125-132
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• 1991

The studies on the carried out to investigate the effects of capacitation method of spermatozoa on the in vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and FCS for 24~48hrs in an incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18hrs with motile capacitated sperm by preincubation of mKRB, treatment of HIS(high strength ion), Ca-IA(Inophore A), BFF(bovine follicular fluids) and heparin. The results obtained in these experiments were summarized as follows : 1. The in vitro fertilizatin and cleavage rate offollicular oocytes fertilized with capacitated spermatozoas in BO solution by preincubation of mKRB, treatment of HIS, Ca-IA, BFF and heparin method were 53.1%, 33.9%, 50.8%, 48.1%, 58.8% and 28.1%, 17.7%, 26.2%, 22.8%, 32.8%, respectively. And the fertilization and cleavage rate of heparin method was of highest of all. 2. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solutin by both caffeine, BSA and heparin methods were 65.8%, 70.3% and 40.8%, 47.3%, respectively, and those rates were higher treatment of heparin＋BSA, heparin＋caffeine than treatment of heparin. 3. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoa in BO solution with heparin concentrations of 2, 5, 10, 20, 40$\mu\textrm{g}$/ml were 50.0%, 54.7%, 58.1%, 51.7% and 27.9%, 32.8%, 37.1%, 30.0%, respectively. And the fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution with 10$\mu\textrm{g}$/ml of heparin was the highest of all. 4. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with caffeine concentraton of 10, 20, 30, 40$\mu\textrm{g}$/ml were 71.4%, 74.3% and 70.6%, 70.0% and 45.7%, 47.3%, 44.1%, 41.4%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with caffeine and heparin together(70.3~74.3%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%). 5. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with BSA concentration of 5, 10, 20, 30$\mu\textrm{g}$/ml were 63.6%, 62.9%, 66.7%, 60.3% and 44.1%, 43.5%, 48.5%, 42.7%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with BSA and heparin together(60.3~66.73%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%).

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.83-93
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• 1997

Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization (MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after partial zona dissection (PZD), subzonal insertion (SUZI), and intracytoplasmic sperm injection (ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not undergo standard IVF-ET repetitively because of failure in fertilization or extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles. From March, 1995 to May, 1996, 27 cycles of IVF-ET with ICSI in 19 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $10.50{\pm}6.13$ in 30 previous cycles, and $10.57{\pm}5.53$ in 27 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was $7.89{\pm}4.30$, and the fertilization rate of $67.9{\pm}20.2%$ could be obtained after ICSI. The number of embryos transferred was $1.43{\pm}2.40$ in previous cycles, and $4.36{\pm}1.77$ with the mean CES of $41.8{\pm}27.4$ in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 29.6% (8/27) per cycle and 42.1% (8/19) per patient with the clinical pregnancy rate of 22.2% (6/27) per cycle and 31.6% (6/19) per patient. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with the past history of failure in fertilization or low fertilization rate of oocytes in the previous IVF-ET cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to effective management of infertile couples.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.105-109
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• 2003

Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

• Korean Journal of Animal Reproduction
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• v.12 no.2
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• pp.112-119
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• 1988

These studies were conducted to investigate the effects of culture temperature and time on the in vitro maturation and semen type and media on the in vitro fertilization of bovine follicular oocytes, and to asses in vitro fertilization rate of oocytes cultured by extraffollicular method following fertilization in vitro, or transfer into the pseudopregnant rabbit oviduct or uterus. The bovine oocytes recovered from follicles were cultrued for 18 hrs or 72hrs at 38$^{\circ}C$ with 5% CO2 in moist air. Flesh-diluted(2 folds) and frozen-thawed semen in 0.5ml straw from a fertile bull were used. In order to obtain capacitation of spermatozoa were treated with bovine follicular fluids(BFF) and Inophore A(IA). The results obtained were summarized as follows: 1. The oocytes were classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 62.0%, 12.0%, 17.2%, 5.9% and 3.0% of the total oocytes harvested, respectively. 2. The oocytes matured to metaphase II were significantly increased between 24-48hrs of incubation and at 37-39$^{\circ}C$ with 5% CO2 in moist air. 3. The in vitro fertilization rate following transferred into rabbit oviduct or uterus with bull semen and in vitro matured oocytes were higher ligation than non-ligation of oviduct or uterus. 4. The in vitro fertilization rate of oocytes matured in vitro were higher neat than frozen semen and treatment of IA than BFF on the capacitation of spermatozoa. 5. The effects of semen types and media on in vitro fertilization of oocytes matured in vitro were higher fertilization rate of neat than friozen semen, and media was not significant.