• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.65-69
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• 1992

The sucess of intrauterine inseminations with washed human spermatozoa was evaluated in 92 patients. In according to indication of insemination, intrauterine inseminations of women with hostile cervical mucus yield a 35.3% and 12.8% with male factors and 27.3% with unexplained infertility. In ovulation induction group with variable agents including clomiphen, human menopausal gonadotropin (HMG) and GnRH analog, the pregnancy rate was 27.8% (22/79) and in natural cycle group, 15.4% (2/13). The fetal loss rate in insemination group was 12.5% (3/24). Multiple pregnancies were 7 cases and ovulation induction were performed in 6 cases among them. Intrauterine insemination with washed human spermatozoa therefore represents an effective and safe procedure selected infertile couples.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.2
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• pp.596-602
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• 2016

Experiments were carried out examine short-term effects of human chorionic gonadotropin (hCG) and long-term effects of luteinising hormone releasing hormone analogue (LHRHa) on milt production of the tongue sole Cynoglossus semilaevis. In the first experiment, each fish was implanted with a blank cholesterol pellet (control), 100 and $200{\mu}g$ LHRHa per kg body weight. In the second experiment, fish were injected with either 100, 200, 400 and 800 IU hCG per kg body weight or same volume of marine fish Ringer's solution. In the first experiment, milt volume was increased in male implanted with $200{\mu}g$ LHRHa pellet compared with other groups at day 10. Injection of 400 and 800 IU hCG resulted in an increase in the milt volume at hour 96 after the treatment. Although statistical difference is unable to confirm because of small milt volume, compared with the control group, hormone pellet-treated groups had a reduction in the mean spermatocrit (Sct) and sperm concentration (Sc). The results suggest that the increase in milt volume is at least partially gonadotropin (GtH)-dependent and increased milt volume has a relationship with milt hydration.

• 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.

• Journal of Embryo Transfer
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• v.23 no.1
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• pp.19-24
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• 2008

The purpose of this study was to compare the efficiency of slow freezing with that of vitrification method for the cryopreservation of human embryos. Human embryos were derived from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and the mixed solution of propanedial (1.5, 1.0, 0.5M PROH) and sucrose (0.1M), ethylene glycol (7.5, 15%), dimethyl sulfoxide (7.5, 15% DMSO), sucrose (0.5, 1.0M) and SPS (Serum Protein Substitute) was used for a cryoprotectant for slow freezing and vitrification solution, respectively. Rates of recovery after thawing, morphological normality, post-thaw viability, arrest, morphological abnormality and preimplantation development were compared between two protocols. After freezing-thawing, recovery and survial rate of slow freezing was (88.6% and 73.4%), whereas vitrification was (99.2% and 96.2%) (p<0.05). The arrest rate of slow freezing was significantly lower compared with those of vitrification(8.7% vs 29.9%) (p<0.05). Preimplantation development to the 2-cell (83.8% vs 67.7%), 4-cell (69.0% vs 47.2%) and 8-cell (62.4% vs 37.8%) stages 24, 48 and 72 h after thawing, respectively, were higher in the slow freezing than the vitrification. After slow freezing and vitrification of human embryo at 2-8cell stage, the rate of recovery rate, survival rate and partial damage rate were 92.0% vs 100%, 80.4% vs 96.2% and 52.2% vs 19.0%, respectively. And partial damage rate was significantly lower than those of slow freezing method (p<0.05). These results demonstrate that a slow freezing using PROH is more efficient than a vitrification for cryopreserving the human zygotes, although the vitrification yielded better recovery, survival and partial damage of frozen-thawed 2-8 cell stage embryos than slow freezing method.

• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.91-96
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• 2013

Transgenic chickens have been spotlighted as an highly potent bioreactor for their fecundity, short generation time, and eggs associated with mass production of protein. In this study, we generated transgenic chickens exhibiting oviduct specific expression of human growth hormone fused to human transferrin for oral administration. Gene of the modified growth hormone located at downstream ovalbumin promoter (~3.6 kb) was introduced to stage X blastodermal cell employing retrovirus vector system. Several transgenic chickens were successfully generated. However, genomic analyses showed unexpected deletion within the transgene. The modification of the transgene seemed to occur during germ cell formation because the deletion was detected only from the sperm DNA of the G0 founder animal. There was no evidence of deletion in the somatic cell DNA samples of the same chicken. Consequently, same pattern of the deletion was confirmed in both somatic and germ cells of the G1 progeny.

• Toxicological Research
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• v.17
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• pp.309-312
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• 2001

It has recently been suggested that the release of "endocrine disrupters (EDs)" into the environment has resulted in adverse health effects on wild life populations and humans (Golden et al., 1998; Tyler et al., 1998; Kang et al., 2000). Human sperm counts have declined significantly throughout the world during the past fifty years, and which is a significant public health concern (Carlsen et al., 1992; Carlsen et al.. 1995). In addition, the EDs persisting in the environment are known to disrupt the normal endocrine systems of wildlife (Colborn, 1995; Crewet al., 1995; Folmer et at, 1996; Sumpter, 1995; Tyler, 1998). Some estrogenic chemicals bind to estrogen receptors (Bolger et al.. 1998), interfere with the binding of physiological ligands to steroid hormone-binding proteins (Danjo, 1997; Milligan et al., 1998). and show immunotoxicity (Sakae et al., 1998). (omitted) (omitted)

• Clinical and Experimental Reproductive Medicine
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• v.45 no.4
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• pp.189-194
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• 2018

Objective: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. Methods: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. Results: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p<0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. Conclusion: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.6
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• pp.754-758
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• 2001

Bovine oocytes with compact and complete cumulus cells were cultured for up to 24h in TCM199 buffered with 25mmol/l hepes and supplemented with 10% FBS (fetal bovine serum), 1mg/ml $17{\beta}$-estradiol, 20IU/ml hCG(human chorionic gonadotropin). All of the oocytes were divided into at 6 groups depending upon incubation times (control, 0 hour, 6 hours, 12 hours, 16 hours, 18 hours). To all experimental media, $200{\mu}g/ml$ puromycin was added at different incubation times mentioned above. Following these culture times, in vitro insemination was conducted with frozen-thawed bovine spermatozoa in medium BO (Brackett and Oliphant medium for in vitro insemination) with $10{\mu}g/ml$ BSA(bovine serum albumin) and 10 mg/ml heparin added. After 22h culture, the oocytes were fixed with acetic alcohol solution and stained with orcein acetic solution to evaluate sperm nuclear progression. Addition of puromycin after 0, 6 and 12 h of culture resulted in near of oocyte maturation at the M1 stage. Contrariwise, puromycin addition after 12 h of culture led to restoration of nuclear progression to M2 stage. On the one hand, puromycin affected the synthesis of Cyclin B protein that may be involved in the oocyte maturation and sperm capacitation for in vitro fertilization. The present study suggests the participation of protein synthesis, cyclin B, in the oocyte development from M1 to M2 stages in vitro.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.40 no.6
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• pp.374-379
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• 2007

Two experiments were designed to examine short-term effects of human chorionic gonadotropin (hCG), and long-term effects of gonadotropin-releasing hormone agonist (GnRHa), $17{\alpha}-hydroxyprogesterone$ (17P), and $17{\alpha},20{\beta}-dihydroxy-4-pregnen-3-one\;(17,20{\beta}P)$, alone or in combination, on milt production of the starry flounder Platichthys stellatus. In the first experiment, fish were injected with either 200 IU hCG/kg body weight or the same volume of marine fish Ringer's solution (MFRS). In the second experiment, each fish was implanted with a blank cholesterol pellet (control), $200\;{\mu}g$ GnRHa, $500\;{\mu}g$ 17P, or $100\;{\mu}g\;17,20{\beta}P/kg$ body weight alone or in combination. In the first experiment, hCG injection resulted in an increase in the expressible milt volume and a decrease in the spermatocrit (Sct). After pellet implantation in the second experiment, the milt volume was increased in males treated with GnRHa, GnRHa+17P, or $GnRHa+17,20{\beta}P$. On day 7 after hormone pellet implantation, the milt volume began to increase, and on day 14, the milt volume in the $GnRHa+500\;{\mu}g$ 17P group was significantly higher than that in the control group. Compared with the control group, the hormone pellet-treated groups had a significant reduction in the mean Sct and sperm concentration (Sc) at day 7 after pellet implantation, while there were no differences in total sperm number. The results suggest that increases in milt volume are generally associated with decreases in Sct and SC, suggesting that the main mechanism for the increase in milt volume was milt hydration.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.319-324
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• 1997

Despite the direct placement of sperm within the oocyte, fertilization failure still occurs after ICSI. This study was accomplished to analyze the chromosomes in oocytes failed to fertilize after ICSI comparing to oocytes failed to fertilize by conventional in vitro insemination. Seventy-four ICSI cycles and 122 conventional IVF cycles were included in analysis. Included unfertilized oocytes were from 74 patients (mean age = $32.7{\pm}3.7$). Ninety-three oocytes were informative and 83 oocytes were legible for cytogenetic analysis. Sixty-two oocytes out of 83 (74.7%) had normal chroruosomes, while 15 (18.1%) were hypoploidy, 6 (7.2%) were hyperploidy. Eighteen oocytes out of 93 (17.6%) were premature chromosome condensation (PCC). Two hundred ninety-four unfertilized oocytes after conventional insemination were subjected to chromosomal analysis and 180 oocytes were legible for analysis. One hundred thirty-two oocytes out of 180 (73.3%) were normal, while 22 (12.2%) were hypoploidy, 20 (11.1%) were hyperploidy, and 6 (3.3%) were polyploidy. Twenty-two oocytes (12.2%) were PCC. There was no difference in chromosomes between oocytes that failed to fertilize after ICSI or conventional insemination. High PCC rates in fertilization-failed oocytes suggest that oocytes maturity is another important factor in achieving successful fertilization.