• Journal of Embryo Transfer
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• v.28 no.3
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• pp.163-168
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• 2013

Embryo transfer (ET) technology is of high importance in modern cattle breeding programs. ET is one step in the process of removing one or more embryos from the reproductive tract of an outstanding donor female and transferring them to one or more recipient females. Embryos also can be produced in the laboratory via techniques such as in vitro fertilization (IVF). But the actual transfer of an embryo is only one step in a series of processes that may include some or all of the following: superovulation and insemination of donors, collection of embryos, isolation, evaluation and short-term storage of embryos, micromanipulation and genetic testing of embryos, freezing of embryos and embryo transfer. Cryopreservation and direct transfer of frozen-thawed embryos is common-place with pregnancy rates near that of fresh embryos. Polymerase chain reaction (PCR) technology is currently being used for sexing embryos, and this technology will be used for "embryo diagnostics" and "embryo genomics" in the future. Although, many limitations and problems remain to overcome, these and other new technologies promise to change livestock breeding drastically in the next decade.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.173-181
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• 1993

In order to demonstrate whether ovoperoxidase hardens the zona of oocytes activated by incubating in M-S buffer supplemented with 20$\mu$M of Ca-ionophore A 23187, the effect of peroxidase inhibitors(250mM pheylhydrazine, 28mM sodilum sulfite, 350mM glycine ethyl ester and 50mM sodium azide), tyrosine analogue(12.5mM tyramine) and exogeneous peroxidase(50$\mu\textrm{g}$/ml horseradishperoxidase ; HRP) on zona hardening in ionorphore-treated oocytes were investigated. The results obtained from thses experiments were summarized as follows : 1. The zona solubility (t50) of ionophore-activated and DMSO-treated oocytes at 1, 2 and 3 hr of culture were 25.0, 31.6 and 40.6min., and 9.7, 10.8 and 15.5 min., respectively. The longest time required for zona lysis of ionophore activated oocytes at 1 hr after onset of ionophore treatment. The diferences int50 for zona was significantly greater as compared to DMSO-treated controls(P<0.01). 2. The inhibition rates of hardening in the oocytes treated with the phenylhydrazine, sodium sulfite, glycine ethyl ester and sodium azide, were 23.8, 61.9, 95.2 and 23.8%l, respectively, and the tyramine, was 14.3%. Several known peroxidase inhibitors and tyrosine analogue were blocked zona hardening in ionophore activated oocytes. 3. The treatment of exogeneous peroxidase promoted the zona hardening of activated oocytes but not unactivated oocytes. These resuls indicate that the ovoperoxidase apparently catalyzes the hardening of the zona following ionophore activation of mouse oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.13 no.1
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• pp.67-75
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• 1986

Follicle monitoring in the normal and clomiphene·stimulated cycles were analyzed in the Seoul IVF and ET (In vitro fertilization and embryo transfer) program. Ovarian follicular diameters were measured by the real·time sector scanner and plasma estradiol levels were assayed by radioimmunoassay methods during periovulatory period. The maximum follicular sizes of the clomiphene-stimulated and normal cycles were 21.1+-3.4mm and 19.2+-0.8mm, respectively. The peak levels of plasma estradiol in the clomiphene-stimulated and normal cycles were 10538+-553.6ng/ml and 298.3+-39.6pg/ml, respectively. Daily growth rate of the follicular diameters of the clomiphene-stimulated and normal cycles were 2.1mm and 1.9mm, respectively. Mean follicular number of the clomiphene-simulated and normal cycles were 2.28+-1.12 and 1.12+-0.21, respectively. There was a good statistical correlation between the mean follicular diameters and the plasma estradiol levels in the normal ovulatory and c1omiphene-stimulated ovulatory menstrual cycles (p<0.05). Our data revealad that the mean follicular diameter and the plasma estradiol level prior to HCG administration in IVF and program should reach at the level of 17.8+-3.0mm and 949.4+-487.1 pg/ml, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.2
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• pp.87-92
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• 2011

Objective: This study aimed to determine the safety and clinical effect of artificial shrinkage (AS) in terms of assisted hatching of fresh blastocysts. Also, we evaluated the correlation between patient age and the effect of AS on clinical outcome. Methods: Two AS methods, using a 29-gauge needle and laser pulse, were compared. Seventy-three blastocysts were shrunk using a 29-gauge needle and the same number of other blastocysts were shrunk by a laser pulse. We evaluated the shrunken blastocysts hourly and considered them viable if they re-expanded >70%. Blastocyst transfer cycles (n=134) were divided into two groups: a control group consisted of the cycles whose intact embryos were transferred (n=100), while the AS group consisted of the cycles whose embryos were replaced following AS (n=34). The implantation and pregnancy rates of the control group and AS group were compared ($p$ <0.05). Results: The re-expansion rates of the 29-gauge needle and laser pulse AS groups were similar (56 [76.7%] vs. 62 [84.9%], respectively). All of the remaining shrunken blastocysts were re-expanded within 2 hours. There was no degeneration of shrunken blastocysts. The total and clinical pregnancy rate of the AS group (23 [67.6%]; 20 [58.8%], respectively) was significantly higher than that of the control group (47 [47.0%]; 39 [39.0%], respectively). In the older patient group, there was no difference in the clinical outcomes between the AS and control groups. Conclusion: These results suggest that AS of blastocoele cavity, followed by the transfer, would be a useful approach to improve the clinical outcome in cycles in which fresh blastocyst stage embryos are transferred.

• YAKHAK HOEJI
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• v.50 no.6
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• pp.409-414
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• 2006

Although mammalian sperms are cryopreserved for in vitro fertilization a process of cryopreservation decreases the fertility. Acrosome reaction requires depolarization-induced Ca$^{2+}$ influx and Ca$^{2+}$ releases from the Ca$^{2+}$ stores. To examine whether the cellular Ca$^{2+}$ mobilization is altered by a sperm cryopreservation we compared cytosolic Ca$^{2+}$ signals between fresh and cryopreserved pig sperms using confocal Ca$^{2+}$ imaging. The magnitudes of depolarization induced Ca$^{2+}$ increases were significantly smaller in cryopreserved sperms. Exposures to 10 mM caffeine or 5 ${\mu}$M thapsigargin elicited less Ca$^{2+}$ increases in the cryopreserved sperms compared to fresh sperms. In addition, progesterone-trig-gered Ca$^{2+}$ rises, that are thought to enhance acrosome reaction, were completely abolished in the cryopreserved sperms. These results suggest that storage and(/or) release of Ca$^{2+}$ from the intracellular Ca$^{2+}$ stores in pig sperms are significantly impaired by the process of cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.3
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• pp.268-272
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• 2021

Bone marrow-derived cell (BMDC) therapy has numerous applications as potential biological cells for use in regenerative medicine. Here, we present an original case of endometrial atrophy associated with genital tuberculosis in a woman who achieved a live birth with BMDC. This 27-year-old woman came to our center with endometrial atrophy and primary infertility. She had a past history of genital tuberculosis and amenorrhea. Her husband's semen quality was normal. The patient was counseled for hysteroscopy due to thin endometrium and advised in vitro fertilization (IVF) with donor eggs in lieu of poor ovarian reserve. Several attempts of IVF with hormone replacement therapy (HRT) were made, but the desired thickness of the endometrium was not achieved. Uterine artery injection of BMDC through interventional radiology was given, followed by HRT for three months, which resulted in improved endometrium. This was subsequently followed by IVF with donor egg. The treatment resulted in the conception and delivery of a 3.1-kg baby boy through lower segment caesarean section with no antenatal, intranatal or postnatal complications. Recently, there has been massive interest in stem cells as a novel treatment method for regenerative medicine, and more specifically for the regeneration of human endometrium disorders like Asherman syndrome and thin endometrium, which was the reason behind using this strategy for treatment.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.273-281
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• 2014

This study was carried out to establish the system of OPU derived embryo production, management of recipients as well as offspring production. OPU derived embryo production system was carried out of aspiration of immature oocytes 2 times per week, total 24 times for 3 months by an ultrasonographic guided follicular aspiration system and then produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture system. This work was collected total 13,866 oocytes, average $8.2{\pm}4.5$ oocytes per session and 8,170 G1 + G2 grade oocytes, average 4.8 oocytes per session by 1,692 times session of total 71 donors for 4 years from 2010 to 2013. The rate of cleavage and blastocyst developmental competence were obtained 11,825 (85.3%) and 5,032 (36.3%) that was $7.0{\pm}3.8$ cleaved embryos and $3.0{\pm}2.5$ blastocysts per session. OPU derived embryo transfer were taken place in 2, 4, 6 and 7 local governments at 2010, 2011, 2012 and 2013 for 4 years and pregnancy rate were obtained 41.2, 43.9, 46.5 and 49.7% in each years. It means that pregnancy rate was continuously improved according of every year for 4 years. Pregnancy rate was significantly different according to individual local government in which was 62.7% in B, but 24.2% in F at 2012. Paternity identification was carried out total 26 offspring in C local government of 2012 and then confirmed 100% agreement of its analysis. In conclusion, the results obtained the possibility of mass production of elite cow embryos as well as offspring by OPU derived embryo production system, of which could be decreased the required time of genetic improvement.

• Development and Reproduction
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• v.15 no.4
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• pp.291-299
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• 2011

The membranous adhesions could induce implantation failure despite transplantation of high quality of embryo. Clinically, of the patients who have membranous filmy adhesions, endometrial polyps have been found in not infrequently. Thus this study was tried to evaluate the features of endometrial polyps and the effect of endometrial polyps on formation and extents of membranous adhesions in uterine cavity of infertile patients under hysteroscopy. A retrospective study was conducted on 34 infertile patients who were diagnosed as endometrial polyps with membranous adhesions during hysteroscopy from July 2008 to July 2011. Number, size, location and morophologic type of endometrial polyps were investigated. If needed, methylene blue solution was instillated to endometrial cavity to identify membranous adhesions. Then, associations between membranous adhesions with features of endometrial polyps were evaluated. Mean size of endometrial polyp was $1.6{\pm}0.6$ cm, the bigger of endometrial polyps was, the larger of extents of membranous adhesions. (p<0.05). Endometrial polyps were locate evenly in endometrial cavity as follows: anterior uterine wall, 39.1%; posterior uterine wall, 34.8%; lateral uterine wall, 26.1%; upper: 29.4%, middle: 32.4%, lower segment, 35.3%. Mean number of endometrial polyps was $2.26{\pm}1.3$. The pedunculated type was 37.7% and sessile type was 32.4%. There was no statistically significant association of location, number and morphologic type of endometrial polyps with membranous adhesions. In conclusion, hysteroscopy before in vitro fertilization on infertile patients was worthy because of removing of endometrial polyps and membranous adhesions.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.123-130
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• 1997

Normal maturation of the mammalian oocytes is prerequisite for the fertilization and the early embryonic development. We have been tested the effects of purine and its de novo synthetic inhibitor, azaserine(Aza) on the maturation of germinal vesicle(GV) and germinal vesicle breakdown(GVBD) mouse oocytes. Denude-immature oocytes were cultivated in the media containing adenosine, guanosine, and/or azaserine, and checked the matruation stage by monitoring the prominent morphological changes. In GV stage oocytes, GV was arrested temporarily by the adenosine(1.0%) and protractedly by the guanosine(65.9%, P<0.001). The regression was increased significantly at the adenosine(90%, P<0.001) but decreased at the guanosine(1.6%, P<0.05). Inhibiting the de novo synthesis of purine, nuclear maturation rate was increase(90.4% : 96.7%), but GV arrest was significantly increased by cotreatment with guanosine(P<0.001). Polar body extraction significantly was increased at the Aza(P<0.05), but not in others. In GVBD oocytes, adenosine itself did not affect GVBD arrest. Guanosine, on the other hand, elevated GVBD arrest rate(P<0.001), but co-treated with Aza, decreased GVBD arrest(P<0.001). Aza increased GVBD arrest rate(20.2%, P<0.05) compared with control. From those results, we know that guanosine shows more prominent effect on the inhibition of nuclear maturation at the GV stage, and of the 1st polar body extrusion at the GVBD stage. Adenosine showed the cytoplasmic toxicity at GV stage oocyte. Our data speculate that cytoplasmic cAMP level is auto-regulated by endogenous adenylate cyclase while GVBD is inhibited by guanosine, since purine toxicity is not observed in the GVBD stage. And it is showed that purine metabolism is concerned with nuclear maturation, that the amounts of purine metabolism is not even during the oocyte maturation.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.7
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• pp.1068-1076
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• 2020

Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa. Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T₀). The diluted specimens were stored at 4℃ for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T₂₄, T₄₈). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system. Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa. Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.