• Journal of Embryo Transfer
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• v.24 no.3
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• pp.145-150
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• 2009

This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.55-62
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• 2010

Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, DC and CDCs. Activity of MMP-2 in the DC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the CDCs 36 hr. Expression of TIMP-3 protein in the CDCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

• Archives of Pharmacal Research
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• v.27 no.11
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• pp.1168-1176
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• 2004

Exposure to stress is known to precipitate or exacerbate many reproductive dysfunctions such as dysmenorrhea and infertility. Abnormalities of the reproductive system, as shown by reduced ovulation, fertilization and early embryonic development, are frequently seen in dysmenorrhea and infertility. It has been generally accepted that Gamisoyosan (GSS) is a useful prescription for treating insomnia, dysmenorrhea and infertility induced by a stress. Also GSS has been used traditionally to improve systemic circulation and biological energy production. Based on these, this study investigates whether GSS improved ovarian dysfunction caused by stress in mice. Mice were subjected to stress by electric shock on the foot for 30 min daily for a week and treated with GSS at 500 / body weight per day for one week. Thereafter, changes body weight, adrenal weight, ovulation rate, in vitro and in vivo fertilization, embryonic development and estradiol concentrations were measured. GSS markedly increased the body weight of mice with stress, but not normal mice. The administration of GSS caused a reduction in adrenal weight in stressed mice. GSS also had significant positive effects on ovulation rate, estradiol production, in vivo and in vitro fertilization rates and embryonic development. These results indicate that GSS can improve the reproductive dysfunctions caused by stress, and these may production biological energy.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.40-46
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• 2017

Objective: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. Methods: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. Results: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. Conclusion: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

• Korean journal of applied entomology
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• v.34 no.2
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• pp.140-146
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• 1995

Changes in protein content during oocyte development was measured and egg-specific protein was characterized from the eggs in Lucilia illustris. During normal development ovarian protein was rapidly increased at 72hr and reached maximum at 96hr after a protein meal, when the eggs were fully matured. Purified protein from the ovaries by gel filtration of DEAE-cellulose an Sephacryl S-200 was loaded on 7.5% native polyacrylamide gel electrophoresis and identified at ${R}_{f}$ 0.4 as egg-specific protein, which has a mol. wt of 110,000. A total of 13 amino acids in th egg-specific protein was identified and expecially asparagine, glutamic acid, and tyrosine were highly concentrated. Five fatty acids were also identified. It is suggested that there is a specific protein in the eggs of L. illustris except yolk protein synthesized and secreted by fat body.

• Development and Reproduction
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• v.2 no.1
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• pp.69-74
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• 1998

The early gonadal development and sexual differentiation of Rhynchocypris oxycephalus are described from the stage of hatching to 150 days after hatching. During this peroid, the average length of the body grew from 0.64 cm to 5.96 cm. the primordial germ cells (PGCs), which could be recognized at the time of hatching, began to protrude into peritoneal cavity at a standard total length of 1.91 cm. At a standard total length of 2.29cm, initial ovarian differentiation wasidentified by the transformation of PGCs to meiotic oocytes. Finally, at the standard total length of 5.96 cm, the female gonads gradually developed towards migratory nucleus oocytes, characterisiing the maturation. Oocytes proliferated rapidly after sex differentiation while the testis entered a period of quiescence, as they continued to multiply but did not undergo growth until the standard total length of 4.00 cm. At a standard total length of 4.00 cm, spermatocytes arrested in thephase of interkinesis, Sertoli-like cells and sperm duct formation, with signs of meiotic activity, were observed. Therefore it may be concluded that R. oxycephalus belongs to the differentiated type of gonochoristic teleosts.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.473-479
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• 1997

The yolk protein of spotted flounder, Verasper variegatus was purified by precipitation wit cold distilled water, followed by Sepharose CL-6B column chromatography. The purified protein was identified as vitellin by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The purified vitellin from ovarian crude extracts has same antigenic determinants with the female specific serum protein, vitellogenin. The molecular weight of purified vitellin was estimated about 550 kD by gel filfration. The vitellin was composed of three major subunits with molecular weight of about 108, 85 and 31 kD, and two minor subunits. The vitellin was identified by western blot analysis using anti-vitellin antibody.

• Journal of Apiculture
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• v.34 no.3
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• pp.245-254
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• 2019

We investigated the anti-tumor effects and molecular mechanism of Brazil, China and Korean regional propolis on HeLa ovarian cancer cell line. Each propolis extracts was prepared by ethanol extraction method. Cytotoxicity of propolis extracts was determinated by EZ-cytox cell viability assay. To necessity of anti-tumor effect and molecular mechanism of propolis, we must be adjusting propolis concentration. Due to 100 ㎍/mL of propolis extract were reduced cell viability to less than 50%, we adjusted all of propolis concentration to 100 ㎍/mL. By Western blotting analysis, we confirmed that anti-tumor mechanism of Brazil, China and Korea regional propolis has significantly difference. All of propolis was activated apoptosis related molecules such as PARP, caspase-3. However, cell proliferation signaling molecules including Akt1, ERK and Bcl-2 were reduced the protein expression level. Especially, the expression of tumor suppressor protein p53 was significantly increased in propolis-treated group such as Gyeonggi, Chungbuk, Chungnam, Jeonbuk, Gyeongnam and China. The phosphorylation of Bax which as apoptosis indicator was appeared in propolis-treated group such as Gyeonggi, Gangwon, Chungnam, Gyeongbuk, China. In this results showed that the regional propolis has completely different mechanism in anti-tumor. Thus, propolis extracts may be useful source of functional materials on anti-cancer and it will be able to choose the suitable propolis for cancer therapy by analyzing individual characteristics.

• Development and Reproduction
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• v.4 no.2
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• pp.187-193
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• 2000

Gonad and blood samples were taken from the cultured female Korean sea bass, Lateolabrax japonicus from October to February between 1997 and 1999. Gonadosomatic index began to increase in November and reached the highest value in December (12.8$\pm$1.5) and January (14.8$\pm$3.5), and then decreased sharply in February (2.6$\pm$1.5, p<0.05). The ovarian oocytes developed to tertiary yolk stage and reached fully-Brown stage in December and January, and then underwent atresia without maturation and ovulation in February. The plasma estradio3-17 $\beta$ level increased from November, and reached the highest value in December (1,152.3$\pm$107.2 pg/ml) and January (1,315.4$\pm$99.5 pg/ml), after then decreased in February (P<0.05). The concentration of plasma 17 $\alpha$ ,20 $\beta$-dihydroxy-4-pregnen-3-one was not significantly changed at low levels (86.6$\pm$6.5∼93.8$\pm$2.8 pg/ml) during the experimental period (P<0.05). All the fish with fully-grown oocytes in the ovary were matured and ovulated by HCG injection. The number of floating eggs were 325,000$\pm$26,000 at HCG 1,000 luhg and 195,000$\pm$35,000 at 2,000 lUikg. There was no difference in fertilization rate and hatching rate of the eggs (P<0.05). Considering these results, we could infer that the ovarian oocyte of the cultured Korean sea bass were not matured and ovulated because of the lack of gonadotropin surge. Moreover, HCG injection could induce oocyte maturation and ovulation in the cultured fish, and the effective dose was 1,000 IU/kg.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.13 no.4
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• pp.125-134
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• 1980

The dovelopment of the gonads, gametogenesis and the reproductive cycle of the topshell, Turbo cornutus Solander, which is one of valuable food animals fom Korean waters were studied by photomicroscophy. The materials were monthly collected from Bangeojin, Jeongjari and Dangweol, all these places being located in the south-eastern part of Korea, for one year from March 1979 to February 1980. Topshell is dioecious and oviparous. Gonad is situated on the surface of liver, which lies posteriorly. The surface of ovary and testis is covered with a fibrous membrane, membrane of connective and muscular fibers and then an outermost layer of simple-columnar epithelial cells which are composed of cuboidal and columnar mucous gland cells. Primordial germ cells develop on the germinal epithelium of ovarian and testicular lobuli which are originated from the fibrous membrane and extend toward hepatic gland. Undifferentiated mesenchymal tissue and pigment granular cells are abundantly distributed between the growing oocytes and spermatocytes in the early development stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and pigment granular cells are considered to be related to the growing of the oocytes and spermatocytes. Early multiplicating oogonium is ca. $10\mu$ in diameter and nucleushaving a central nucleolus is ra. $8\mu$. As the oocytea grow to ca. $50-60\mu$ by the increase of cytoplasm, the oocytes become look like bunches of grapes which are attached to ovarian lobuli. Mature eggs are ca. $180-210\mu$ in diameter and it is surrounded by a gelatinous membrane of ca. $10\mu$ in thickness. After spawning, undischarged ripe eggs and spermatozoa remain in the ovary and testis respectively for some time. Then they finally degenerate, and proliferation of new oogonia and spermatogonia occur along the germinal epithelia of newly developed ovarian and testicular lobuli. Reprocuctive cycle of Turbo cornutus could be classified into five successive stages: multiplicative, growing, maturer spent and recovery stages. Spawning occurs from August to November with Peak spawning from early September to late October.