• Title/Summary/Keyword: cell culture model

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Development of Predictive Growth Model of Listeria monocytogenes Using Mathematical Quantitative Assessment Model (수학적 정량평가모델을 이용한 Listeria monocytogenes의 성장 예측모델의 개발)

  • Moon, Sung-Yang;Woo, Gun-Jo;Shin, Il-Shik
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.194-198
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    • 2005
  • Growth curves of Listeria monocytogenes in modified surimi-based imitation crab (MIC) broth were obtained by measuring cell concentration in MIC broth at different culture conditions [initial cell numbers, $1.0{\times}10^{2},\;1.0{\times}10^{3}\;and\;1.0{\times}10^{4}$, colony forming unit (CFU)/mL; temperature, 15, 20, 25, 37, and $40^{\circ}C$] and applied to Gompertz model to determine microbial growth indicators, maximum specific growth rate constant (k), lag time (LT), and generation time (GT). Maximum specific growth rate of L. monocytogenes increased rapidly with increasing temperature and reached maximum at $37^{\circ}C$, whereas LT and GT decreased with increasing temperature and reached minimum at $37^{\circ}C$. Initial cell number had no effect on k, LT, and GT (p > 0.05). Polynomial and square root models were developed to express combined effects of temperature and initial cell number using Gauss-Newton Algorism. Relative coefficients of experimental k and predicted k of polynomial and square root models were 0.92 and 0.95, respectively, based on response surface model. Results indicate L. monocytogenes growth was mainly affected by temperature and square root model was more effective than polynomial model for growth prediction.

Analysis of two-stage Continuous Culture System by Transient Response of Single-stage Continuous Culture System (일단 연속 생물반응기의 과도상태 거동을 이용한 이단 연속 생물반응기의 해석)

  • 박성훈;공인수
    • KSBB Journal
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    • v.7 no.4
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    • pp.308-316
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    • 1992
  • Two-stage continuous culture system has been studied intensively to maximize the productivity of a cloned gene product in unstable recombinant microorganism. As an effort to optimize the two-stage process, transient behavior of the second-stage was studied theoretically as well as experimentally using Escherichia coli Kl2$\delta$Hl$\delta$trp. A mathematical model describing the transient response to a step change in dilution rate was developed based on the assumption that the adaptation rate of cell growth is proportional to the available growth potential, which is defined as the difference in dilution rates between before and after shift-up. The kinetic parameters appearing in the model equations were the dimensionless step increase in growth rate($\alpha$) and the adaptation rate constant(k). These parameters were evaluated for various dilution rates and temperatures by washout method. This relatively simple adaptation model could predict the specific growth rate of the second-stage successfully. Advantage and disadvantage of the proposed model are also discussed.

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Optimization of Culture Medium for Novel Cell-Associated Tannase Production from Bacillus massiliensis Using Response Surface Methodology

  • Belur, Prasanna D.;Goud, Rakesh;Goudar, Dinesh C.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.199-206
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    • 2012
  • Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

The Design and Fabrication of μCCA-μGI Device for Toxicity Evaluation of Acetaminophen (아세트아미노펜 독성평가를 위한 μCCA-μGI 디바이스의 개발)

  • Chang Jung-Yun;Shuler Michael L.
    • Journal of Pharmaceutical Investigation
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    • v.36 no.4
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    • pp.263-269
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    • 2006
  • Deficiencies in the early ADMET(absorption, distribution, metabolism, elimination and toxicity) information on drug candidate extract a significant economic penalty on pharmaceutical firms. Microscale cell culture analogue-microscale gastrointestinal(${\mu}CCA-{\mu}GI$) device using Caco 2, L2 and HEp G2/C3A cells, which mimic metabolic process after absorption occurring in humans was used to investigate the toxicity of the model chemical, acetaminophen(AAP). The toxicity of acetaminophen determined after induction of CYP 1A1/2 in Caco 2 cells was not significant. In a coculture system, although no significant reduction in viability of HEp G2/C3A and L2 cells was found, approximately 5 fold increase in the CYP 1A1/2 activity was observed. These results appear to be related to organ-organ interaction. The oral administration of a drug requires addition of the absorption process through small intestine to the current ${\mu}CCA$ device. Therefore, a perfusion coculture system was employed for the evaluation of the absolution across the small intestine and resulting toxicity in the liver and lung. This system give comprehensive and physiologic information on oral uptake and resulting toxicity as in the body. The current ${\mu}CCA$ device can be used to demonstrate the toxic effect due to organ to organ interaction after oral administration,

Experimental Study of Chungganhaeju-tang (Qingganjiejiu-tang) on Oxidative Stress (청간해주탕(淸肝解酒湯)의 항산화 작용에 관한 실험적 연구)

  • Lee, Ji-Eun;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.32 no.2
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    • pp.188-202
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    • 2011
  • Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.

Mathematical Model for Adsorption of Berberine on Encapsulated Adsorbent (캡슬에 고정화된 흡착제에의 Berberine의 흡착에 관한 수학적 모델)

  • 최정우;조상원이원홍
    • KSBB Journal
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    • v.10 no.4
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    • pp.358-369
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    • 1995
  • A mathematical model using local thermodynamic equilibrium isotherms for adsorption in encapsulated adsorbent is proposed in order to optimize the design parameters in situ bioproduct separation process. The model accurately follows the experimental data on the adsorption of berberine, secondary metabolite produced in Thaictrum rugosum plant cell culture. The adsorption rate on encapsulated adsorbent is compared with that on alginate-entrapped adsorbent. The result shows that the higher loading capacity in encapsulated adsorbent is mainly due to the increase in the maximum solid phase concentration. Based on the adsorption rate and loading capacity, the encapsulated adsorbent would be more useful than the entrapped adsorbent when used in situ bioproduct separation process. Design parameters in situ bioproduct separation process, such as the size of the capsule, membrane thickness, the ratio of capsule volume to bulk volume, the ratio of single capsule volume to total capsule volume and the adsorbent content in the capsule, are evaluated by using the model. The ratio of single capsule volume to total capsule volume is the most effective parameter for adsorption of berberine on encapsulated adsorbent.

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Effect of Silver sulfadiazine on the Skin Cell Proliferation and Wound Healing Process in Hairless Mouse 2nd degree Burn Model (설파디아진은의 피부세포 증식 및 화상모델에 있어서의 상처치유과정에 미치는 영향)

  • Cho, Ae-Ri
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.113-117
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    • 2002
  • Cyto-toxic effect of silver sulfadiazine (Ag-SD) on keratinocytes and its implication on wound healing process were investigated in $2^{nd}$ degree bum hairless mouse model. As a dermal model, HaCat (immortalized keratinocytes) monolayer culture in DMEM with 10% FBS was used. Cyto-toxicity of Ag-SD was estimated by measuring the cell viability using neutral red assay after adding the drug. The $2^{nd}$ degree bum was prepared on hairless mouse back skin (1 cm diameter) and dressings with Ag-SD were applied for 96 hr. The process of re-epithelialization and the presence of inflammatory cells were investigated and histology with Hematoxylin-Eosin staining was performed. Ag-SD displayed highly cyto-toxic effect on cultured HaCat cells in a concentration dependent manner $(1-100\;{\mu}g/mL)$. Topical application of Ag-SD (2%) could control the infection: no inflammatory cells were observed in histology. However the cyto-toxic effect of Ag-SD on skin cells induced the impairment in epidermal regeneration.

Phytoremediation of Selected Explosives in a Model System of Plant Tissue Cultures

  • Vanek, Tomas;Nepovim, Ales;Zeman, Svatopluk
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.395-399
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    • 2000
  • The phytoremediation of trinitrotoluene, nitroglycerine, pentaerytritoltetranitrate in plant tissue cultures of Solanum aviculare, Rheum palmatum and Populus simonii were studied. All above mentioned explosives were degradated to to less toxic products and finally mineralized or bound to the cell wall.

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Follistatins have potential functional role in Porcine Embryogenesis

  • Kim, Dong-Hee;Chun, Ju Lan;Lee, Ji Hye;Kim, Keun Jung;Kim, Eun Young;Lee, Bo Myeong;Zhuang, Lili;Kim, Min Kyu
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.52-60
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    • 2016
  • In animal reproduction, the quality of oocytes and embryos has been evaluated by the expression of specific molecules. Follistatin (FST), which was isolated from follicular fluid, binds and bio-neutralizes the TGF-${\beta}$ superfamily members. Previous studies using the bovine model showed FST could be an important molecular determinant of embryo developmental competence. However, the effect of FST treatment on porcine embryo developmental competence has not been established. In this study, the effect of exogenous FST on porcine embryo developmental competence was investigated during in vitro culture. FST (10 ng/ml) treatment induced a significant decrease in the rate of cell arrest at the 4-cell stage. The expression levels of DNA-methyltransferase 1 (DNMT1), histone deacetylase 1 (HDAC1), and histone deacetylase 2 (HDAC2) were decreased in 4-cell stage embryos. FST treatment also resulted in significant improvements in developmental competence of embryos in terms of blastocyst formation rate and OCT-4 mRNA levels, the latter being related to pluripotency. In conclusion, during in vitro culture, FST treatment significantly ameliorated 4-cell block during embryonic development and improved embryo developmental competence. Therefore, FST treatment may potentially have a functional role in porcine embryogenesis that is broadly applicable to enhance in vitro embryo development.