• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.168-173
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• 2004

The ability of frozen-thawed fetal skin was examined to generate viable cell lines for nuclear transfer. Fetal skin frozen at -20$-20^{\circ}C$, $-30^{\circ}C$ or $-80^{\circ}C$ in the presence of 5% DMSO used as tissue explants to generate somatic cells. The resultant confluent cells were then used as donors for nuclear transfer (NT). Of the bovine NT embryos reconstracted from the somatic cells, 62.3%, 76.6% to 65% showed cleavage 70.5%, 81.9% to 78.5% reached the stage of morula formation and 39.7%, 43.2% or 47.6% reached the blastocyst stage. There was no significant difference in development when the NT embryos were compared with those reconstracted from fresh somatic cell derieved skin tissues (72%, 75.3%, and 45.2%, for cleavage, and development to morula and blastocyst stage, respectively). NT embryos were then placed in a portable $CO_2$ incubator and carried to China from Japan by air. After reaching to farm, two NT embryos were transferred to each of 5 recipients. We obtained 2 NT calves which birth weights is 30kg and 36kg female, and gestation periods is 281 and 284 days, respectively. There were no observation any abnormality from those calves. The results indicated that cell lines derieved from bovine fetal skin cryopreserved by a simple method could be used as donors in nuclear transfer using the portable $CO_2$ incubator.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.141-145
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• 2012

The present study was conducted to develop a simple method for porcine oocyte maturation without $CO_2$ regulation. In experiment 1, we evaluated that the effect of $CO_2$ non-supplement on porcine oocyte maturation. Cumulusoocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three groups (Control, tube-$CO_2$, and tube-non-$CO_2$). For control, COCs were cultured in 4-well multidish in a $CO_2$ incubator. For tube-$CO_2$, COCs were cultured in a round-bottom tube in a $CO_2$ incubator, and for tube-non-$CO_2$, COCs were cultured in a round-bottom tube sealed tightly without $CO_2$ supplement in a dry incubator. The proportion of oocytes reached to metaphase II (M-II) was not significantly different among three groups (87.9% to 91.4%). In experiment 2, we evaluated the effect of $CO_2$ non-supplement during oocyte maturation on development of embryos. Oocytes with a polar body were divided into two groups (Control and tube-non-$CO_2$) and applied 1.1 kV/cm or 1.2 kV/cm voltages for parthenogenetic activation. After activation, embryos were cultured for 6 days and examined the development. The proportion of embryos cleaved was not significantly different among treatment (86.3% to 91.5%). The proportion of embryo reached to blastocyst stage was not significantly different among treatment (13.9% to 25.2%). The cell number of blastocysts was not significantly different among treatment (29.0 to 32.4). In conclusion, oocytes cultured in a dry incubator without $CO_2$ supplement have enough competence to development after parthenogenetic activation. These results would be useful for transporting oocytes or embryos a long distance.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.29 no.7 s.238
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• pp.970-975
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• 2005

This paper presents a novel microgenerator of $CO_2$ (carbon dioxide) gas. $NaHCO_3$ (sodium bicarbonate) in a chamber is decomposed by the underlaid microheater. Alternatively, water droplet is caged by paraffin layer and released by heating. The released water dissolve HOC(COOH)$(CH_2COOH)_2$ (citric acid) powder and then, $NaHCO_3$ reacts with the solubilized HOC(COOH)$(CH_2COOH)_2$ and $CO_2$ is produced. Micropumps actuated by $CO_2$ generation were fabricated. A portable micro cell incubator of which pH is controlled by the produced $CO_2$ is also presented as one of the further applications.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.210-221
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• 1995

The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.189-196
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• 2019

Objective: We aimed to evaluate the effects of different oxygen conditions (20% [high O₂], 5% [low O₂] and 5% decreased to 2% [dynamic O₂]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture. Methods: The high-O₂ and low-O₂ groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O₂ group, mouse embryos were cultured from the one-cell to the morula stage under 5% O₂ for 3 days, followed by culture under 2% O₂ to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets. Results: The blastocyst formation rate was significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The total cell number was significantly higher in the dynamic-O₂ group than in the low-O₂ and high-O₂ groups. Additionally, the apoptotic index was significantly lower in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. Conclusion: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.

• Journal of IKEEE
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• v.27 no.1
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• pp.122-126
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• 2023

In this paper, a control system for a complex microbial incubator was proposed. The proposed control system consists of a control unit, a communication unit, a power supply unit, and a control system of the complex microbial incubator. The controller of the complex microbial incubator is designed and manufactured to convert analog signals and digital signals, and control signals of sensors such as displays using LCD panels, water level sensors, temperature sensors, and pH concentration sensors. The water level sensor used is designed and manufactured to enable accurate water level measurement by using the IR laser method with excellent linearity in order to solve the problem that existing water level sensors are difficult to measure due to foreign substances such as bubbles. The temperature sensor is designed and used so that it has high accuracy and no cumulative resistance error by measuring using the thermal resistance principle. The communication unit consists of two LAN ports and one RS-232 port, and is designed and manufactured to transmit signals such as LCD panel, PCT panel, and load cell controller used in the complex microbial incubator to the control unit. The power supply unit is designed and manufactured to supply power by configuring it with three voltage supply terminals such as 24V, 12V and 5V so that the control unit and communication unit can operate smoothly. The control system of the complex microbial incubator uses PLC to control sensor values such as pH concentration sensor, temperature sensor, and water level sensor, and the operation of circulation pump, circulation valve, rotary pump, and inverter load cell used for cultivation. In order to evaluate the performance of the control system of the proposed complex microbial incubator, the result of the experiment conducted by the accredited certification body showed that the range of water level measurement sensitivity was -0.41mm~1.59mm, and the range of change in water temperature was ±0.41℃, which is currently commercially available. It was confirmed that the product operates with better performance than the performance of the products. Therefore, the effectiveness of the control system of the complex microbial incubator proposed in this paper was demonstrated.

• Korean Journal of Veterinary Research
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• v.57 no.2
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• pp.89-95
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• 2017

This study was conducted to determine the effects of biophoton treatment during in vitro maturation (IVM) and/or in vitro culture (IVC) on oocyte maturation and embryonic development in pigs. An apparatus capable of generating homogeneous biophoton energy emissions was placed in an incubator. Initially, immature pig oocytes were matured in the biophoton-equipped incubator in medium 199 supplemented with cysteine, epidermal growth factor, insulin, and gonadotrophic hormones for 22 h, after which they were matured in hormone-free medium for an additional 22 hr. Next, IVM oocytes were induced for parthenogenesis (PA) or provided as cytoplasts for somatic cell nuclear transfer (SCNT). Treatment of oocytes with biophoton energy during IVM did not improve cumulus cell expansion, nuclear maturation, intraoocyte glutathione content, or mitochondrial distribution of oocytes. However, biophoton-treated oocytes showed higher (p < 0.05) blastocyst formation after PA than that in untreated oocytes (50.7% vs. 42.7%). In an additional experiment, SCNT embryos produced from biophoton-treated oocytes showed a greater (p < 0.05) number of cells in blastocysts (52.6 vs. 43.9) than that in untreated oocytes. Taken together, our results demonstrate that biophoton treatment during IVM improves developmental competence of PA- and SCNT-derived embryos.

• Journal of Periodontal and Implant Science
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• v.26 no.3
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• pp.681-688
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• 1996

Periodontal regeneration requires the migration and proliferation of gingival fibroblasts and periodontal ligament cells. These cellular events are influenced and regulated by growth factors and some drugs. The purpose of this study is to examine the effect of Rhizoma coptidis and Centella asiatica extracts on human gingival fibroblasts. Gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with ${\alpha}-MEM$ at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator for 2 or 3 days, as a measure of cell proliferation potential, it was examined that the DNA synthesis using $[^3H]-thyrnidine$ incorporation, the cell numbers (with or without dye), and cell viabilities. Rhizoma coptidis is increased the proliferation of gingival fibroblasts at concentration of $10^{-9}g/ml$, but Centella asiatica is decreased the proliferation at all concentrations. This study demonstrated that Rhizoma coptidis is a potential mitogen for human gingival fibroblasts in vitro, and we can expect the usefulness of this drug in periodontal regeneration.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.3
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• pp.43-50
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• 2007

Background and Objectives : The aim of this study was to investigate the anti-oxidant and anti-inflammatory effects of the Taglisodog-eum(TSE) extract on the RAW264.7 cell Methods : The RAW264.7 cell was cultured using Dulbecco's modified Eagle's medium(DMEM, USA), including the 10% fetal-bovine serum(FBS; Sigma, USA) in a $37^{\circ}C$, 5% CO2 incubator. Results : The anti-oxidant ability of TSE were dose-dependantly increased. The LPS-induced IKK, iNOS and COX-2 mRNA expression were dose-dependantly decreased in the RAW264.7 cells treated with TSE. $NF-kB$ activation was suppressed. Conclusion : The findings in this study show that TSE has anti-oxidant and anti-inflammatory effects, such as the inhibition of $NF-kB$ activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.3 no.1
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• pp.19-26
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• 1970

During the summer of 1969, a series of experiments on the spawning and development of the eggs of grass carp and silver carp was performed with the following results, but the complete development of eggs Iras not accomplished: 1. The season of maturity for both species is estimated to be from the end of June to the beginning of August. 2. Pituitary glands for the use of inducing maturation should be obtained before the maturation season of its donor. 3. Silver carp as small as 40 cm in body length (48 cm in total length) with an age of 6 years were found to have reached maturity, and the number of eggs in the ovaries were from about 23 to 26 thousand for those with body lengths of 40-44.5 cm. 4. The conical-shaped net cloth incubator worked better than others, and each incubator was most effective when installed in parallel series to the water supply pipe with a valve for each incubator to control water flow. 5. During the egg development, if any cell or cells were displaced, the eggs eventually died. 6. The proper manipulation of brood fish before the egg-taking seems to be very Important, and close care should be paid to feeding them well and keeping them in a pond of proper water depth.