• Asian-Australasian Journal of Animal Sciences
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• 제17권2호
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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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• 제36권3호
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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.

• 대한기계학회논문집A
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• 제29권7호
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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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• 제25권2호
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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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• 제46권4호
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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.

• 전기전자학회논문지
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• 제27권1호
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• pp.122-126
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• 2023

본 논문에서는 복합 미생물 배양기의 제어시스템을 제안하였다. 제안하는 제어시스템은 복합 미생물 배양기의 제어부, 통신부, 전원부, 제어시스템 등으로 구성된다. 복합 미생물 배양기의 제어부는 아날로그 신호와 디지털 신호의 변환, LCD 패널을 이용한 디스플레이, 수위센서, 온도센서, pH 농도센서 등과 같은 센서들의 신호 제어를 하도록 설계 및 제작한다. 사용하는 수위센서는 기존 수위센서가 거품과 같은 이물질 등으로 인해 측정이 어려운 문제점을 해결하고자 직진성이 우수한 IR 레이저 방식을 사용하여 정확한 수위 측정이 가능하도록 설계 및 제작한다. 온도센서는 열 저항 원리를 사용하여 측정함으로써, 높은 정확도와 누적 저항 오차가 없도록 설계하여 사용한다. 통신부는 2개의 LAN 포트와 1개의 RS-232 포트로 구성하여 복합 미생물 배양기에서 사용되는 LCD 패널, PCT 패널, 로드셀 컨트롤러 등의 신호를 제어부에 전달할 수 있도록 설계 및 제작한다. 전원부는 제어부와 통신부가 원활하게 동작할 수 있도록 24V, 12V 5V 등 3개의 전압 공급 단자로 구성하여 전원을 공급하도록 설계 및 제작한다. 복합 미생물 배양기의 제어시스템은 PLC를 사용하여 pH 농도센서, 온도센서, 수위센서 등의 센서값과 배양에 사용되는 써큘레이션 펌프, 써큘레이션 밸브, 로터리 펌프와 인버터 로드셀 등의 동작을 제어한다. 제안된 복합 미생물 배양기의 제어시스템의 성능을 평가하기 위하여 공인인증기관에서 실험한 결과는 수위 측정감도의 범위가 -0.41mm~1.59mm로, 물 온도의 변화 폭이 ±0.41℃로 현재 상용으로 판매되는 제품들 성능보다 우수한 성능으로 동작됨이 확인되었다. 따라서, 본 논문에서 제안한 복합 미생물 배양기의 제어시스템의 효용성이 입증되었다.

• 대한수의학회지
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• 제57권2호
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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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• 제26권3호
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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.

• 한방안이비인후피부과학회지
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• 제20권3호
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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.

• 한국수산과학회지
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• 제3권1호
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• pp.19-26
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• 1970

1969년 여름 부산수산대학 양어장에서 1963년 일본으로부터 도입하여 길러온 초어와 백련을 사용 채란과 부화실험을 하였는 바, 그 결과는 좋지 못하였지만, 다음과 같은 점을 지적한다. 1. 부산지방의 못에서 양식한 초어가 백련의 성숙 채란 적기는 6월 말부터 8원 초까지의 사이에 있다고 인정된다. 2. 산란 촉진용 뇌하수체는 동종 또는 근연종의 친어로부터 그들의 산란기이 진에 적출 보관하였다가 사용하는 것이 좋다고 인정된다. 3. 백련은 체장 40cm(전장 48cm) 가량의 작은 개체도 6년생은 모두 채란 가능하였으며, 포란수는 체장 $40\~44.5cm$ 되는 것에서 $23\~26$ 만개로 추산되었다. 4. 부화 시설로서는 원추형 망부화기를 급수 파이프에 병렬하고, 각 부화기에는 수량 조절용 발브를 달아서 수중에 설치하는 것이 좋은 동작을 하였다. 5. 알의 발생 중, 분할된 할구(세포)가 배체 밖으로 방출되는 것이 생기면, 그 알은 예외없이 괴사하였다. 6. 친어의 사전 관리를 보다 철저히 하는 것이 긴요함을 느꼈으며, 좋은 먹이와 적절한 수심 및 수온을 유지시키도록 해야 할 것이다.