• Korean Journal of Animal Reproduction
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• v.16 no.1
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• pp.7-14
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• 1992

These experiments were designed to investigate the effects of glycosaminoglycans(GAGs), which maybe produced by cumulus cells, on in vitro maturation process of bovine follicular oocytes. The rates of in vitro maturation of bovine oocytes were examined after incubating with the various concentrations of a hyaluronic acid, chondroitin sulfate, or heparin for 26 hours. The results obatained in these experiments were summarized as follows : 1. When the cumulus or removed oocytes treated with hyaluronic acid(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml), the maturation rates of cumulus-removed oocytes were 48.6, 59.4, 68.4 and 61.3% respectively. Especially, at a concentration of 800$\mu\textrm{g}$/ml, the maturation rates(68.8%) of cumulus-removed oocytes were slightly lower than that(78.3%) of cumulus-enclosed oocytes. However, the treatment of hyaluronic acid showed significantly higher maturation rates compared to that(48.4%) of control group of cumulus-removed oocytes(p<0.05). Therefore, these results suggest thate hyaluronic acid had a beneficial effect on maturation of bovine follicular oocytes. 2. The maturation rate of cumulus-free oocytes treated with chondroitin sulfate(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml) was 52.3, 56.1, 55.9 and 52.2% respectively. These rates were not different from those(52.9%) of control group. However, these rates in cumulus-free oocytes were significantly lower than that(79.0%) in cumulus-enclosed oocytes(p<0.01). Therefore, these results suggest that chondroitin sulfate do not affect on the maturation of oocytes. 3. In cumulus-free oocytes cultured with different heparin concentration, the maturation rates were ranged from 48.5 to 52.1%, showing no differencies from that(50.7%) of control group. However, these rates were significantly lower than 80.0% in cumulus-enlosed oocytes(p<0.01). 4. The nuclear maturaton of oocytes was increased by treatment of each of three glycosaminoglycans(hyaluronic acid, chondroitin sulfate, and heparin). In addition, the treatment of mixed together showed the significant additive effect. Hyaluronic acid was more effective than chondroitin sulfate and heparin were.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.162-167
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• 2021

Camel (camelus dromedarius) is a unique large mammalian species that can survive harsh environmental conditions and produce milk, meat, and wool. Camel reproduction is inferior when compared to other farm animal species such as cattle and sheep. Several trials have been reported to increase camel reproduction and production through assisted reproductive techniques (ARTs) such as in vitro fertilization and cloning. For these reasons, obtaining enough mature oocytes is a cornerstone for ARTs. This demand would be improved by the oocyte in vitro maturation (IVM) systems. In this review, the current approaches and views from different laboratories using ARTs and the IVM to produce embryos in vitro in camel species. For the last two decades, conventional IVM system was the common approach, however, recently the bi-phasic IVM system has been introduced and showed promising improvement in IVM of camel oocytes. Detailed studies are needed to understand camel meiosis and IVM to efficiently increase the production of this species.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.242-242
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• 2004

This study was conducted to investigate the effects of various supplements added into maturation medium of immature porcine oocytes on quantity of cytoplasmic lipid droplets(LD), subsequent fertilization and development to the blastocyst stage in vitro. The basic maturation medium was TCM 199 + 1 ㎍/㎖ FSH, 0.57 mM cystein, 10 ng/㎖ EGF and was supplemented various supplements(10% FBS, 10% pFF, 0.4% BSA, 1.0% BSA, 0.4% PVP, 1.0% PVP). (omitted)

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.139-146
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• 2017

The objective of this study was to determine the effect of fructose that was supplemented to a chemically defined in vitro maturation (IVM) medium on oocyte maturation and embryonic development after parthenogenesis in pigs. The base medium for in vitro maturation (IVM) was porcine zygote medium (PZM) that was supplemented with 0.05% (w/v) polyvinyl alcohol (PVA) or 10% (v/v) porcine follicular fluid (pFF). In the first experiment, when immature pig oocytes were matured in a chemically defined medium that was supplemented with 5.5 mM glucose or with 1.5, 3.0 and 5.5 mM fructose, 3.0 mM fructose resulted in a higher nuclear maturation (91.5%) than 1.5 and 5.5 mM fructose (81.9 and 81.9%, respectively) but showed a similar result with 5.5 mM glucose (94.2%). However, there was no significant differences among groups in the embryo cleavage (89.4-92.4%), blastocyst formation (37.5-41.1%), and mean cell number of blastocyst (30.8-34.2 cells). Fructose at the concentration of 3.0 mM (1.08 pixels/oocyte) resulted in a higher intra-oocyte glutathione (GSH) content than 1.5 and 5.5 mM fructose (1.00 and 0.87 pixels/oocytes, respectively) while the cumulus cell expansion was not influenced. In the second experiment, effect of individual and combined supplementation of a chemically defined maturation medium with 5.5 mM glucose or 3.0 mM fructose was examined. No significant effect was found in the nuclear maturation (86.3-92.6%). Embryo cleavage was significantly increased by the combined supplementation with glucose and fructose (95.2%) compared to that with 3.0 mM fructose only (85.7%) while blastocyst formation (37.3-42.8%) and embryonic cell number (33.3-34.1 cells) were not altered. Effect of supplementation of pFF-containing medium with glucose and fructose + glucose was examined in the third experiment. No significant effect by the supplementation with glucose and fructose or glucose alone was observed in the nuclear maturation of oocytes (90.7-94.1%) and blastocyst formation (51.0-56.5%). Our results demonstrate that 3.0 mM fructose was comparable to 5.5 mM glucose in supporting in vitro oocyte maturation and embryonic development after parthenogenesis and could be used as an alternative energy source to glucose for in vitro maturation of pig oocytes.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.125-131
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• 2009

The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.309-317
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• 2017

Melatonin (N-acetyl-5-methoxytryptamine) is an indole synthesized from tryptophan by the pineal gland in animal. The major function of melatonin is to modulate circadian and circannual rhythms in photoperiodic mammals. Importantly, however, melatonin is also a free radical scavenger, anti-oxidant, and anti-apoptotic agent. Recently, the beneficial effects of melatonin on oocyte maturation and embryonic development in vitro have been reported in many species such as pig, cattle, sheep, mouse, and human. In this review, we will discuss recent studies about the role of melatonin in the production of porcine and bovine oocytes and embryos in vitro in order to provide useful information of melatonin in oocyte maturation and embryo culture in vitro.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.75-80
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• 2003

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.39-46
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• 2000

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50${\mu}{\textrm}{m}$ $\alpha$-tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100${\mu}{\textrm}{m}$ cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

• Development and Reproduction
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• v.12 no.2
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• pp.125-132
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• 2008

Melatonin (N-acetyl-5-methoxytryptamine), a major hormone of pineal gland in vertebrates, is known to be associated with regulation of the dynamic physiological functions in general and has some functions on reproduction in the ovarian follicles in particular. And its antioxidant properties as a scavenger are also reported. The aim of this study was to investigate the effect of melatonin on the in vitro maturation of mouse germinal vesicle (GV)-stage oocytes. Oocyte maturation, apoptosis, and mRNA expression of melatonin receptor were analyzed in the cumulus cell-enclosed oocytes (CEOs) cultured with melatonin for 18 h. The CEOs were obtained from 3 wk-old ICR female mice cultured in media with 0, 0.1 nM, 10 nM, or 1,000 nM melatonin for 18 h. And then the extrusion of the first polar body was assessed to evaluate the maturation rate. The apoptosis and mRNA expression of melatonin receptor (Mtnr1-a and Mtnr1-b) in cumulus cells of each group were measured by TUNEL assay, ELISA, and real time RT-PCR after in vitro maturation(IVM). The addition of melatonin in the IVM medium significantly improved nuclear maturation of the mouse GV oocytes and the highest maturation rate were obtained from the group treated with 1,000 nM melatonin. Apoptosis was not detected in IVM oocytes, but detected in cumulus cells. And cumulus cells treated with 1,000 nM melatonin exhibited significantly lower apoptosis. In the group treated with 1,000 nM melatonin, the expression of melatonin receptor mRNA was decreased in CEOs. In conclusion, melatonin has a potentially important role for regulating oocyte maturation and reduces the apoptosis of cumulus cells in vitro.

• Korean Journal of Veterinary Research
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• v.32 no.1
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• pp.135-142
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• 1992

This experiment was carried out to establish an effective technique of in vitro maturation of porcine follicular oocytes. Porcine ovaries were collected from an abbatoir and delivered to the laboratory in phosphate buffered saline in an hour. Immatured follicular oocytes were collected from the ovaries and divided into groups by the size of follicles and by the attachment of granulosa cells. The follicular oocytes were cultured in m-KRB solution supplemented with FCS(10%), follicular fluid(10%) or hormones of PMSG(10IU/ml), hCG(10IU/ml ) and $estradiol-17{\beta}(1{\mu}g/ml)$ for 48 hours at $39^{\circ}C$ under an atmosphere of 5% $CO_2$ in air. The results are as follows ; 1. The mean recoveration rate of follicular oocytes was 61.8%. 2. The maturation rate was significantly(p<0.05) higher when the oocytes were collected from large-sized follicles and under good state of granulosa cell attachment. 3. The maturation rate was significantly(p<0.01) promoted when the follicular oocytes were cultured in m-KRB solution supplemented with follicular fluid(74.8%) or hormones and fetal calf serum(70.6%).