• Title/Summary/Keyword: 고정화 세포

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Development of Porcine Pericardial Heterograft for Clinical Application (Microscopic Analysis of Various Fixation Methods) (돼지의 심낭, 판막을 이용한 이종이식 보철편의 개발(고정 방법에 따른 조직학적 분석))

  • Kim, Kwan-Chang;Choi, Chang-Hyu;Lee, Chang-Ha;Lee, Chul;Oh, Sam-Sae;Park, Seong-Sik;Kim, Woong-Han;Kim, Kyung-Hwan;Kim, Yong-Jiin
    • Journal of Chest Surgery
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    • v.41 no.3
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    • pp.295-304
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    • 2008
  • Background: Various experimental trials for the development of bioprosthetic devices are actively underway, secondary to the limited supply of autologous and homograft tissue to treat cardiac diseases. In this study, porcine bioprostheses that were treated with glutaraldehyde (GA), ethanol, or sodium dodecylsulfate (SDS) were examined with light microscopy and transmission electron microscopy for mechanical and physical imperfections before implantation, Material and Method: 1) Porcine pericardium, aortic valve, and pulmonary valve were examined using light microscopy and JEM-100CX II transmission electron microscopy, then compared with human pericardium and commercially produced heterografts. 2) Sections from six treated groups (GA-Ethanol, Ethanol-GA, SDS only, SDS-GA, Ethanol-SDS-GA and SDS-Ethanol-GA) were observed using the same methods. Result: 1) Porcine pericardium was composed of a serosal layer, fibrosa, and epicardial connective tissue. Treatment with GA, ethanol, or SDS had little influence on the collagen skeleton of porcine pericardium, except in the case of SDS pre-treatment. There was no alteration in the collagen skeleton of the porcine pericardium compared to commercially produced heterografts. 2) Porcine aortic valve was composed of lamina fibrosa, lamina spongiosa, and lamina ventricularis. Treatment with GA, ethanol, or SDS had little influence on these three layers and the collagen skeleton of porcine aortic valve, except in the case of SDS pre-treatment. There were no alterations in the three layers or the collagen. skeleton of porcine aortic valve compared to commercially produced heterografts. Conclusion: There was little physical and mechanical damage incurred in porcine bioprosthesis structures during various glutaraldehyde fixation processes combined with anti-calcification or decellularization treatments. However, SDS treatment preceding GA fixation changed the collagen fibers into a slightly condensed form, which degraded during transmission electron micrograph. The optimal methods and conditions for sodium dodecylsulfate (SDS) treatment need to be modified.

Determination of Optimum Bead Size by Calculating Effectiveness Factors in Cyclosporin A Fermentation by Immobilized Cells (Cyclosporin A 고정상 발효에서 효율인자의 계산을 통한 고정화 담체의 최적크기 결정)

  • 전계택;이태호장용근
    • KSBB Journal
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    • v.11 no.1
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    • pp.30-36
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    • 1996
  • Based on fermentation data for cyclosporin A production, simple Monod kinetics was proposed for both immobilized and suspended cultures. Higher value of $\mu$mas and lower value of Km suggest better catalytic activity of the immobilized cells than the parallel suspended cells. Furthermore, lower Km value in the immobilized cell system indicates higher affinity of the immobilized cells for carbon substrate as compared with the suspended cells. For immobilized cell cultures, these parameters were also utilized for the estimation of effectiveness factor, an indicator for intraparticle mass transfer resistance. Based on simulation studies, optimum radius of celite beads was turned out $100 ~ 500{\mu}m$In this simulation work, we examined the influence of biosupport size and immobilized biomass density on diffusional resistance of substrate inside the bead matrix. In order to maintain uniformly distributed cell activities in biosupport, it was essential to determine optimum slze of particle, and then to estimate the most economic loaded biomass content.

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Hollow Fiber Membrane Bioreactor (실관 막 생물 반응기)

  • Kim, In Ho
    • Applied Chemistry for Engineering
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    • v.5 no.6
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    • pp.911-916
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    • 1994
  • Hollow fiber membrane has been successfully developed as an artificial kidney device in the 1970's. In the early 1970's animal cells were introduced into a hollow fiber membrane cartridge and well propagated in the cartridge. Since then, hollow fiber membrane was utilized as a bioreactor in order to immobilize enzymes as well as to culture microbial cells and plant cells. In this review, the present status and the prospect of hollow fiber membrane bioreactor are investigated in view of cell density and product productivity.

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Immobilization of Kluyveromyces marxianus FO43 for Ethanol Production (Kluyveromyces marxianus FO43의 Algiante 고정화와 에탄올 발효특성)

  • Lee, Hee-Suk;Shin, Ji-Hyun;Choi, Eon-Ho
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.20-25
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    • 1995
  • This experiment was attempted to improve ethanol productivity by immobilization of Kluyveromyces marxianus FO43 using Jerusalem artichoke powder. Sucrose medium was used to determine optimum conditions for cell immobilization. The optimum conditions were alginate concentration of 2%, bead size of 2 mm, a particle input ratio of 30 : 100, cultivation period of 24 hours, and substrate concentration of 10%(w/v). The immobilized cells produced the high concentrations of ethanol at pH $4.5{\sim}6.5$ and $30{\sim}45^{\circ}C$, broader ranges of pH and temperatures than those of free cells. Under optimum conditions the immobilized cells showed ethanol concentration of 46.4 g/L and productivity of 1.93 g/L.h. The microphotograph using a two phase contrast microscope showed that immobilized cells cultivated under the optimum conditions were densely populated toward the surface area of beads.

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Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.576-587
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    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.

Electrochemical Degradation of Phenol by Using Reticulated Vitreous Carbon Immobilized Horseradish Peroxidase (Horseradish Peroxidase가 고정화된 다공성 탄소 전극을 이용한 페놀의 전기화학적 분해)

  • Cho, Seung-Hee;Yeon, Kyeong-Ho;Kim, Gha-Young;Shim, Joon-Mok;Moon, Seung-Hyeon
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.12
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    • pp.1263-1269
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    • 2005
  • Horseradish peroxidase, had the phenol degradation rate of 95% in aqueous phase, was covalently immobilized on the surface of reticulated vitreous carbon(RVC) and the degradation of phenol was performed with in situ generated $H_2O_2$-immobilized HRP complex in an electrochemical reactor. The incorporation of carboxylic group on the RVC surface was confirmed by FT/IR spectrometry and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC) was used for peptide bonds between the carboxylic groups on the RVC surface and amine groups from HRP. The optimal conditions of in situ $H_2O_2$ generation such as concentration($10{\sim}200$ mM) and pH($5.0{\sim}8.0$) of electrolyte, supply of $O_2(10{\sim}50$ mL/min) and applied voltage($-0.2{\sim}-0.8$ volt, vs. Ag/AgCl) from potentiostat/galvanostat were determined by concentration of hydrogen peroxide and current efficiency. It was observed that the RVC immobilized HRP was stable maintaining 89% of the initial activity during 4 weeks. The phenol degradation rate of 86% was attained under the optimal condition of in situ $H_2O_2$ generation.

Immobilization of Yeast Pichia stipitis for Ethanol Production (에탄올 생산을 위한 효모 Pichia stipitis의 고정화)

  • Lee, Sang-Eun;Lee, Ji-Eun;Kim, Eun-Jin;Choi, Joon-Ho;Choi, Woon-Yong;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • Journal of Life Science
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    • v.22 no.4
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    • pp.508-515
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    • 2012
  • In this study, DEAE-cotton [derivatized by 2-(diethylamino)ethyl chloride hydrochloride (DEAE HCl)] was prepared as a carrier for immobilized $Pichia$ $stipitis$ for ethanol production. When cotton was derivatized with 0.5 M DEAE HCl, the yeast cell suspension was adsorbed at 100% of the initial cell $OD_{600}$. The adsorbed yeast cells were estimated to be 101.8 mg-dry cells/g-DEAE-cotton. In particular, when a flask culture using the immobilized yeast cells was conducted in a glucose and xylose-containing medium, the yeast cells on the DEAE-cotton gradually produced ethanol, according to glucose and xylose consumption; the ethanol yield was approximately 0.33 g-ethanol/g-monosaccharide. Because DEAE-cotton was successfully used as a carrier for ethanol production from a glucose and xylose-containing medium, we expect that this bioethanol production process may be used for the bioethanol production process from the hydrolysate of lignocellulosic biomass. All the results of DEAE-cotton were compared with those of DEAE-cellulose as a carrier for immobilization.

The Roles of Lipid Supplements in Ethanol Production Using a Continuous Immobilized and Suspended Cell Bioreactor (연속식 고정화 및 현탁 세포 생물 반응기에 의한 에탄을 생성중 지질 첨가 영향)

  • Gil, Gwang-Hoon
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.1-8
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    • 1996
  • A one-stage, continuous-flow bioreactor with both immobilized and suspended cells was used to investigate the roles of lipid supplements in ethanol production by Saccharomyces cerevisiae. The reactor performance and the level of alcohol dehydrogenase(ADH) activities of the suspended cells, grown under various conditions, were measured. When ergosterol and/or oleic acid were added with surfactants to the yeast culture grown under non-aerated conditions, remarkable increases in ethanol production and cell growth was achieved, but specific ADH activities were not affected. Especially, no difference of specific ADH activities of the suspended cells grown under aerated and non-aerated condition was observed. The addition of the surfactant as a supplement also resulted in significant increases in ethanol production, cell growth, and specific ADH activity. When ergosterol and oleic acid were added to the yeast culture exposed to higher ethanol concentration($>40\;g/{\ell}$) level, ethanol production, cell growth, and specific ADH activity were increased, but the addition of surfactant was as effective as at lower ethanol concentration level. The results indicated that lipid supplements were more effective at higher ethanol concentration level than at lower ethanol concentration level during ethanol production. ADH isozyme patterns of the yeast cultures grown under various conditions on starch gel electrophoresis showed only one major band, probably ADH I. The migrating distance of the major isozyme, however, varied slightly according to the culture conditions of the cells. No apparent correlation was found between specific ADH activity and amount of ethanol produced. Cell mass was more important factor for ethanol production than specific ADH activity of the cells.

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Sorbitol production from Jerusalem artichoke by inulinase and permeabilized Zymomonas mobilis (Inulinase와 투과성이 향상된 Zymomonas mobilis를 이용한 Jerusalem artichoke로 부터의 sorbitol생산)

  • 김인철;전억한
    • KSBB Journal
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    • v.7 no.1
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    • pp.15-20
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    • 1992
  • The use of Jerusalem artichoke containing $\beta$-1, 2-fructose oligomer in the production of sorbitol that is used as food additives and precursor for the L-sorbose has been studied. Coimmobilization of both inulinase and oxidoreductase was considered for the simultaneous reaction for hydrolysis of inulin and conversion of glucose and fructose liberated from inulin to sorbitol. Both inulinase and oxidoreductase were immobilized in chitin(5%, w/v) and K-carrageenan(4%, w/v), The activity of oxidoreductase was specified by permeabilization of Zymomonas mobilis cell with 0.2% CTAB(Cetyltrimethylammonlumbromide). The use of inulinase for hydrolysis of inulin resulted in 36.65g/l of glucose and 85.32g/1 of fructose respectively. These are valuable substrates for sorbitol production. Using these hydrolyzates, accumulation of 35.64g/l for sorbitol occurred at $38^{\circ}C$ and pH6.2. When permeabilized cells and inulinase were coimmobilized, sorbitol produced at 30.15g/l although it is low compared with 35.64g/l in separated reactor system.

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RGD Fixation of Film Surface and Synthesis of Copolymer Comprising Malic acid (Malic acid 함유 공중합체의 합성 및 필름표면의 RGD 고정화)

  • 이찬우;송경헌
    • Polymer(Korea)
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    • v.26 no.3
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    • pp.300-306
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    • 2002
  • To study the effect of polymer compositions on the cell adhesion, copolymers of 3-(S)-[(dodecyloxycarbonyl) methyl]-1,4-dioxane-2,5-dione (DMD) and L-lactide were made, where DMD was synthesized form L-malic acid (L-MA) and glycolic acid. Furthermore, the copolymerization of DMD and L-lactide was performed using tin(II) octanoate as a catalyst. As a result of fixing RGD on the copolymer films, the cell adhesive peptide was fixable on the surface of the film. It was found out that the amount of fixation of RGD also increases by the increase in the amount of MA unit introduction. Since it is gradually decomposed over a long period and neither remains nor accumulation occurs, glycolic acid-$\beta$-dodecylmalate -lactic acid (D-PGML) is greatly expected as a potential biomaterial with improved slow degradability.