• 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 Animal Reproduction
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• v.12 no.3
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• pp.132-140
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• 1988

These experiments were carried out to examine the development capacity of mouse blastomers separated from 2 to 8-cell stage mouse embryos. The female ICR and C3H mice were subjected to supervolution by intraperitoneal injection of PMSG and HCG and then mated with males of the same strain. Embryos were flushed from oviducts and uteri on a proper time after injection of HCG. After removal of zona pellucida with 0.5% pronase, each embryos were separated into 1/2, 1/4, 2/4, 1/8, 2/8 and 4/8 embryos by pipetting or a fine glass needle in Ca2＋$.$Mg-2＋ free Hoppe& Pitts medium containing 0.02% EDTA. Splitted embryos were cultured in Hoppe & Pitts medium for 48h to 72h. The embryos developed to blastocyst were transferred to recipients on 2 or 3 days of pseudopregnancy. On the other hand, a monozygotic pairs of 1/2 embryos developed to blastocyst after 48h in vitro culture were transferred to recipients on 2 days of pseudopregnancy or pregnancy. The results obtained were summarized as follows. 1. Success rates of separation of blastomeres from 2-, 4- and 8-cell embryos were 91.7%, 68.5-92.4% and 60.8-90.6%, respectively. 2. Development rates of various type of blastomeres to blastocyst after 72h in vitro culture were ranged 64.7-87.1%. 3. Blastocysts obtained after 48h in vitro culture were transferred to recipients on 2 or 3 days of pseudopregnancy. The production rates of live fetuses after transfer on 2 days, only 1/2, 2/4 and 4/8 embryos, were 13.2%, 13.5% and 17.2%, respectively and those of embryos transferred on 3 days were 11.8%, 9.6% and 11.5%, respectively. However, the production rates of live fetuses 1/2 embryos following 72h in vitro culture and transfer to recipients on 2 or 3 days of pseudopregnancy were 7.7% and 12.5%, respectively. 4. From 29 and 31 pairs of 1/2 embryos transferred to recipients on 2 days of pseudopregnancy or pregnancy, 4 sets of monozygotic twins were produced from only pregnant recipients.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.89-95
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• 2008

The present study was undertaken to identify changes of plasminogen activators (PAs) in porcine oviductal epithelial cells (POECs) during the estrous cycle classified with post-ovulatory stages (Post-Ov), early to mid-luteal stages (Early-mid L) and pre-ovulatory (Pre-Ov) stages. The urokinase-type plasminogen activator (uPA) was only observed on day 5 and day 7 of culture in the POECs on all the estrous cycles and gradually increased according to increasing culture times, but not Early-mid L. In POECs-conditioned medium, uPA, tissue-type (tPA) and tPA-PA inhibitor (tPA-PAI) activity were observed at all culture times during estrous cycles. The uPA activity of POECs-conditioned medium on Post-Ov stage were significantly (p<0.05) decreased during prolonged cultures. On the other hand, the tPA activity of POECs-conditioned medium at Post-Ov stage was significantly (p<0.05) higher on day 5 than compared to the other days. Although was higher on day 1 at Post-Ov stage, the tPA-PAI activity of POECs-conditioned medium was significantly (p<0.05) higher on day 7 at all stage than that of day 5 of the culture. Taken together, these results suggest that uPA, tPA and tPA-PAI are produced by POECs, and the variations of the PAs activity are regulated in the different stages of the estrous cycle.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• The Journal of the Korea Contents Association
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• v.18 no.9
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• pp.535-543
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• 2018

The traditional culture, which is a historical product possessed by each country, represents the competitiveness of the country and becomes a national resource. Raw lacquer has a very long history and tradition in China. Lacquer, which is the basic material of Raw lacquer. Also it is a natural paint which has the functions of preservation, insectproof and waterproof used from ancient times in Asia, not only in China but also in Korea and Japan. Such Raw lacquer using lacquer has a long history and it is becomes to a tradition. On the other hand, Its value and meaning are being faded by oilpaintings and watercolors. Much effort and research are needed to become more popular. I would like to introduce the lacquering technique in illustration as one way. There are a lots of expression techniques in the present illustration, but lacquer expression is rarely seen. In the illustration, lacquering technique is a modern reinterpretation of traditional culture by combining modern technology with traditional culture. In this paper, a Raw lacquer with a long tradition of soul is informed to the public and it is a way to bring attention to the beauty of lacquer and reproduce the original expression technique as an illustration to enhance the understanding of traditional culture, I hope that Raw lacquer will be used in the field.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1117-1123
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• 2009

This study investigated the effects of IGF-I and EGF on the development of blastocysts or hatched blastocysts during the in vitro culture of embryos from immature porcine oocytes. After the in vitro maturation and fertilization of cumulus-oocyte complexes (COCs) and their culture in vitro in PZM3 medium, we examined the embryo development rate for 168 h. When different concentrations of IGF-I (0, 1, 10, 20 ng/ml) were supplemented to fertilized porcine embryos in vitro, there were no significant differences in cleavage rate, blastocyst development rate or blastocyst hatching rate among the treated groups. On the other hand, when different concentrations of EGF (0, 1, 10, 20 ng/ml) were supplemented to the in vitro culture medium, blastocyst development rate was highest in the group in which EGF was not supplemented and, specifically, it was higher than in the 20 ng/ml treatment group (p<0.05). When 10 ng/ml IGF-I and 1 ng/ml EGF were supplemented separately or simultaneously, there were no significant differences among the treated groups in blastocyst hatching rate and the number of cells in each condition. This study demonstrated that the addition of IGF-I and EGF into PZM3 medium did not enhance development of the blastocyst stage and total cell number in blastocysts.

• Proceedings of the Korea Contents Association Conference
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• 2009.05a
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• pp.915-920
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• 2009

In terms of development of intelligent information devices, the most important element is the function which is able to deliver the information user wants to input on the device without complexities. By using the mean such as our voice, expression, gesture and physical contact we use in our daily routine, we are able to interact with device easily and simply. In this paper, we are initiated to present interactive surface system, in which allows users to use their hand gesture as a function of mouse and keyboard free, to be interacted within the platform of web application.

• KSBB Journal
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• v.31 no.4
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Journal of Aquaculture
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• v.11 no.1
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• pp.113-118
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• 1998

This study was carried out to investigate feeding of bacteria by Tigripus japonicus (Copepoda : Harpacticoida) under axenic culture. The ovigerous females and nauplii were grown with feed of aquatic bacteria. Growth of RT bacteria strain was suppressed by feeding of co-existing T. japonicus. T. japonicus of non-axenic culture was observed with oil bead in the egg sac. On the other hand, early nhauplius stage did not develop to the next stage without stage took bacteria as food. And the adult of T. japonicus may utilize the baxteria as nutrient source for egg development.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.548-553
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• 1999

A production of $\beta-Carotene$was attempted in a fed-batch culture of Blakeslea trispora by controlling both foam and dissolved oxygen in the presence of surfactant, Span 20. Results obtained from the shake flask cultures indicated that a high concentration of dissolved oxygen was needed for both cell growth and $\beta-Carotene$ synthesis, and the optimal concentration of glucose was found to be in the range of 50-100 g/l. In order to maintain the dissolved oxygen concentration level at higher than 50% of air saturation, pure oxygen was automatically sparged into the medium with air. Foam was controlled by bypassing air from the submerged aeration to the headspace in response to the foam that was caused by Span 20. High agitation speed was found to be detrimental to the cell growth due to shear damage, even though it provided sufficient dissolved oxygen. On the other hand, a low aeration speed caused stagnant regions in the fermentor because of improper mixing. Thus, for the fed-batch operation, agitation speed was increased gradually from 300 to 700 rpm to prevent cell damage at the initial stage of fermentation and to give efficient mixing for a viscous culture broth as the culture proceeded. By controlling dissolved oxygen and foam, a high concentration of $\beta-Carotene$otene (1,190 mg/l) was obtained in 6 days of the fed-batch culture of B. trispora with 2.5% of the dry cell weight, which was approximately 5 times higher than that of the batch cultures.