• Title/Summary/Keyword: cell culture techniques

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A STUDY ON THE CYTOTOXICITY OF ROOT CANAL ANTISEPTIC SOLUTIONS (근관소독제의 세포독성에 관한 연구)

  • Kim, Jae-Gu;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.18 no.1
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    • pp.95-102
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    • 1993
  • Two functions of root canal medicaments and irrigants are to reduce microorganisms and to encourge the repair of apical tissues. The biocompatibility of endodontic materials has been tested using in vitro cell culture techniques. The purpose of this study Was to evaluate and compare the cytotoxic effects of 2 root canal irrigation solutions and 4 antiseptics on HEp-2 and McCoy cells. Two irrigation solutions were sodium hypochlorite. $H_2O_2$ and 4 antiseptics were povidone, ethanol, glutaraldehyde and benzalkonium chloride. Each solutions were serially diluted to 1:1, 1:10, 1:$10^2$, 1:$10^3$, 1:$10^4$, 1:$10^5$, 1:$10^6$. And each diluted solutions were added to the cells and cytotoxic effects were measured with the absorbance of formazan formed cells by ELISA READER. The results were as follows : 1. Benzalkonium chloride was the most cytotoxic on HEp-2 cell. (P<0.05) 2. $H_2O_2$ was the most cytotoxic on McCoy cell. (P<.05) 3. Povidone and ethanol showed mild cytotoxic effect on HEp-2 and McCoy cell. (P<0.05).

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Rapid Detection Method for Human Rotavirus from Vegetables by a Combination of Filtration and Integrated Cell Culture/Real-Time Reverse Transcription PCR (Filtration과 Integrated Cell Culture/Real-Time Reverse Transcription PCR 기법을 이용한 채소류에서 Human Rotavirus 신속 검출)

  • Hyeon, Ji-Yeon;Chon, Jung-Whan;Song, Kwang-Young;Hwang, In-Gyun;Kwak, Hyo-Sun;Lee, Jung-Soo;Kim, Moo-Sang;Lee, Jung-Bok;Seo, Kun-Ho
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.117-123
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    • 2011
  • The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.

Identification of Gene Locus by the Somatic Cell Hybridization in Chicken (체세포 융합에 의한 닭의 유전인자구명에 관한 연구)

  • 정익정
    • Korean Journal of Poultry Science
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    • v.16 no.1
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    • pp.1-8
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    • 1989
  • This experiment was conducted to improve the performance of chickens by the precise separation and analysis of chromosomes which are integrated genetic materials, and by the use of gene manipulation techniques. Following are the main results obtained. 1. When the chromosomes were separated through the leucocyte culture and analyzed by Giemsa banding techniques (especially by the method in which 20 layers of banding patterns could be found in chromosome #1), the normal Patterns of chromosomes #l-9 and sex chromosomes, and the location of constitutive heterochromatin without any gene activities in all chromosomes were discovered. 2. To utilize the primodial germ cells (PGC) as the genetic vector which is one of the most important gene manipulation techniques, PGC's from triploid were transplanted to normal host embryos. Since the donor PGC's(3n) were found in the gonads of growing host embryos gene manipulation in poultry using PGC's, seemed to be possible.

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Mass Production of Poly(3-Hydroxybutyrate) by Fed-Batch Cultures of Ralstonia eutropha with Nitrogen and Phosphate Limitation

  • Ryu, Hee-Wook;Cho, Kyung-Suk;Kim, Beom-Soo;Chang, Yong-Keun;Chang, Ho-Nam;Shim, Hyun-Joo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.751-756
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    • 1999
  • For mass production of poly(3-hydroxybutyrate) (PHB), high cell density cultures of Ralstonia eutropha were carried out in 2.5-1 and 60-1 fermentors by two fed-batch culture techniques of nitrogen and phosphate limitation. When the nitrogen limitation technique was employed using both an on-line glucose monitoring and control system, a high concentration level of PHB (121g/l) was obtained in the small-scale fermentor of 2.5 1. However, the PHB concentration obtained in a large-scale fermentor of 60 1 only turned out to be 60g/l. In contrast, when another fed-batch culture technique of the phosphate-limitation employing dissolved oxygen (DO) stat glucose feeding was used, a large amount of PHB was successfully produced in both 60-1 and 2.5-1 fermentors. In a 2.5-1 fermentor, concentrations of PHB and cells obtained in 58 h were 175 and 210 g/l, respectively, which corresponded to the PHB productivity level of 3.02 g/l/h. In a 60-1 fermentor, a final cell concentration of 221 g/l and a PHB concentration of 180 g/l with PHB productivity level of 3.75 g/l/h were obtained in 48h. PHB content and yield from glucose were 81% and 0.38g PHB/g glucose, respectively. These data suggest that the phosphate limitation technique is more effective compared to nitrogen limitation in the mass production of PHB by R. eutropha of a large scale.

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Selection of Suitable Varieties of Carnation (Dianthus caryophyllus L.) and Optimization of Culture Conditions for Efficient Tissue Culture (효율적 조직배양체계 확립을 위한 카네이션 품종 선발 및 배양조건 설정)

  • Kang, Chan-Ho;Han, Bum-So;Han, So-Gon;Kown, Sung-Hwan;Song, Young-Ju
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.121-129
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    • 2011
  • As the molecular breeding was progressed, many plant transformation techniques were attained for improving transformation accuracy and used to produce useful transgenic plants. Day by day, new varieties were developed so new transformation techniques required for these newly developed varieties. Carnation (Dianthus caryophyllus L.) is a popular and economically important ornamental plant, all over the world. Keeping this in view, we selected 18 varieties of D. caryophyllus L. commonly available in the market and did optimization of culture conditions for more efficient tissue culture and to get higher number of plants via micro-propagation. Four varieties namely Yellowdotcom, Jakarta, Belmonte, Polartessino etc. were selected for organ culture studies from single cell line. The optimum growth was recorded in the MS media supplied with sucrose 3%, NAA 1.0 mg/L and TDZ 1.0 mg/L. except Belmonte, in which, BA 1.0 mg/L was found to be the best combination, in place of TDZ, rest ingredients were same. The most efficient coagulating agent used to obtain higher number of plant from callus was phytagel 0.3%. The most effective explant for higher shoot formation was stem in which 80.2% shoot formation was recorded. It also reduced culture periods by 6 days.

Development of In Vitro Culture System for Male Germline Stem Cells in Porcine (돼지 웅성 생식선 줄기세포의 체외배양기법 개발)

  • Kim, Yong-Hee;Kim, Byung-Gak;Lee, Yong-An;Kim, Bang-Jin;Kim, Ki-Jung;Lee, Myeung-Sik;Im, Gi-Sun;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.171-177
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    • 2009
  • Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from $10{\sim}14$-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and $O_2$ tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, $37^{\circ}C$ temperature, and $O_2$ 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.

The Characteristics of Magic Realism Expressed in the Costume of Tarsem Singh Movies - Focusing on "The Cell" and "The Fall" - (타셈 싱(Tarsem Singh) 영화의상에 표현된 매직리얼리즘의 표현특성 - 더 셀(The Cell)과 더 폴(The Fall) 영화를 중심으로 -)

  • Yang, Soohyun;Lee, Younhee
    • Journal of the Korean Society of Costume
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    • v.65 no.1
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    • pp.1-14
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    • 2015
  • Magic realism, which originated in the culture of Central and South America, creates a fantastic fictional word by linking unrelated incidents using magical. The purpose of this study was to analyze Tarsem Singh's design characteristics that are based on magic realism in his movies, "The Cell" and "The Fall", and to highlight the artistic values appearing in the films. The research was conducted by observing the characteristics and concept of magic realism based on literature and preceding research, and discovering how Singh expressed these creatively and experimentally within the costumes in his films. The results of the study were as follows: Firstly, magical, legendary and symbolic characteristics appear within the fantasy of magical realism, and common techniques within film costuming include repetition of similar objects, solid silhouettes within scenes, and various decorative materials used for fantastic expression. Secondly, regarding ideality, the destruction of previous ideas and recreation of the present were found with materials and details used in film costuming to destroy previous ideas. Expressions of character through external decoration and depiction of living things as not living were also found. Thirdly, reiteration showed the coexistence of history and legend, reality and fantastic elements, and arrangement of opposing elements.

AN IN VITRO STUDY ON CELLULAR RESPONSE OF SEVERAL DENTURE BASE RESINS (수종 의치상 레진의 세포반응에 관한 연구)

  • Jun Chul-Oh;Vang Mong-Sook
    • The Journal of Korean Academy of Prosthodontics
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    • v.30 no.2
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    • pp.247-257
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    • 1992
  • The present study quantitates the in vitro cytotoxicity of a variety of denture base acrylic resins using cell culture techniques combined with image analysis to measure nuclear area and DNA contents. In this study, a comparison was made among direct curing, heat curing and microwave curing resins. The results obtained from this study were as follows : 1. Morphologically, cell process and nucleus became prominent but macroscopic difference according to the resins were nit observed. In addition, increased cellular density around the specimen were observed. 2. In DNA contents measurements, $S-G_2M$ phase cell was 15.47%, 14.58% in control and heat curing resin on 1st day and the others group $21.39\sim33.36%$ were measured. 3. Nuclear area and DNA contents were increased on 3rd day except DNA content of the microwave curing resin group. These results suggest that denture base acrylic resins stimulate gingival fibroblasts in vitro, especially stimulation of direct curing resin is larger and longer than the others.

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Isolation, Culture, and Fusion of Nicotiana Protoplasts (원형질체 분리, 배양 및 Nicotiana 종간 세포융합에 관한 연구)

  • 윤경은;김준철;최상수;손세호
    • Journal of the Korean Society of Tobacco Science
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    • v.1 no.2
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    • pp.138-149
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    • 1979
  • For the preliminary study on tobacco cell fusion as one of new breeding techniques, the conditions that would be most effective in isolation, fusion, and culture of tobacco protoplasts were examined ; 1. The enzyme solution of 0.5% macerozyme and 2% cellulase( or meicellase) was the most economic and efficient in isolating protoplasts from tobacco leaves. 2. The proper incubation period of tobacco leaves in cell wall digesting solution was 4 hours. 3. As an osmotic stabilizer, sorbitol or mannitol solutions were employed. The concentration of 0.5~0.7 M of either hexitol gave satisfying results as the osmotic stabilizer. 4. The calcium concentration appeared to be an important factor in protoplast fusion. The adhesion of protoplasts was enhanced by enrichment of calcium ion in PEG solution. The highest frequency of protoplast fusion was obtained when tobacco protoplasts were incubated in PEG solution. containing 9mM CaCl2. 5. Cell divisions of the isolated protoplasts were continued and have generated colonies when they were grown on B-5 medium at 28$^{\circ}C$.

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Clinical Application of Adipose Derived Stromal Cell Autograft for Wound Coverage (배양하지 않은 지방조직세포를 이용한 창상피복)

  • Seo, Dong-lin;Han, Seung-Kyu;Chun, Kyung-Wook;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.35 no.6
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    • pp.653-658
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    • 2008
  • Purpose: Skin and soft tissue defect is one of the major challenges faced by plastic surgeons. Adipose derived stromal cells, which can be harvested in large quantities with low morbidity, display multilineage mesodermal potential. Therefore, adipose derived stromal cells have been met with a great deal of excitement by the field of tissue engineering. Recently, Adipose derived stromal cells have been isolated and cultured to use soft tissue restoration. In order to apply cultured cells for clinical purpose, however, FDA approved facilities and techniques are required, which may be difficult for a clinician who cultures cells in a laboratory dedicated to research to utilize this treatment for patients. In addition, long culture period is needed. Fortunately, adipose derived stromal cells are easy to obtain in large quantities without cell culture. The purpose of this study is to present a possibility of using uncultured adipose derived stromal cells for wound coverage. Methods: Seven patients who needed skin and soft tissue restoration were included. Five patients had diabetic foot ulcers, 1 patient got thumb amputation, and 1 patient had tissue defect caused by resection of squamous cell carcinoma. The patients' abdominal adipose tissues were obtained by liposuction. The samples were digested with type I collagenase and centrifuged to obtain adipose derived stromal cells. The isolated adipose derived stromal cells were applied over the wounds immediately after the wound debridement. Fibrin was used as adipose derived stromal cells carrier. Occlusive dressing was applied with films and foams and the wounds were kept moist until complete healing. Results: One hundred to one hundred sixty thousand adipose derived stromal cells were isolated per ml aspirated adipose tissue. All patients' wounds were successfully covered with the grafted adipose derived stromal cells in a 17 to 27 day period. No adverse events related to this treatment occurred. Conclusion: The use of uncultured adipose derived stromal cells was found to be safe and effective treatment for wound coverage without donor site morbidity.