• Journal of Biosystems Engineering
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• 제39권3호
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• pp.244-252
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• 2014

Purpose: The development of an efficient in vitro cell culture device to process various cells would represent a major milestone in biological science and engineering. However, the current conventional macro-scale in vitro cell culture platforms are limited in their capacity for detailed analysis and determination of cellular behavior in complex environments. This paper describes a microfluidic-based culture device that allows accurate control of parameters of physical cues such as pressure. Methods: A microfluidic device, as a model microbioreactor, was designed and fabricated to culture Saccharomyces cerevisiae and Chlamydomonas reinhardtii under various conditions of physical pressure stimulus. This device was compatible with live-cell imaging and allowed quantitative analysis of physical cue-induced behavior in yeast and microalgae. Results: A simple microfluidic-based in vitro cell culture device containing a cell culture channel and an air channel was developed to investigate physical pressure stress-induced behavior in yeasts and microalgae. The shapes of Saccharomyces cerevisiae and Chlamydomonas reinhardtii could be controlled under compressive stress. The lipid production by Chlamydomonas reinhardtii was significantly enhanced by compressive stress in the microfluidic device when compared to cells cultured without compressive stress. Conclusions: This microfluidic-based in vitro cell culture device can be used as a tool for quantitative analysis of cellular behavior under complex physical and chemical conditions.

• 한국동물학회지
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• 제35권3호
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• pp.295-307
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• 1992

In order to study the differentiation of the epithelial cells during the development of fetal rat lung tissue, histological changeB in organotypic culture and in vivo were examined. Light microscopy and scanning electron microscopy were used to analvre the histological change in rat lung from the 15th nary of gestation to the 111th nary after birth. In organotypic culture system, the pulmonary epithelial cell differentiation was studied by scanning electron microscopy. The results obtained from this study were as follows. 1. During deveiopment of lung, the glandular stage lasted from the Isth day to the lsth naut of gestation; the canalicular stage from the 17th nay to the 19th naut of gestation; the saccuiar stage from 20th nary to the birth. Alveolar stage was observed at the 3rd nary of postnatal rat lung. 2. In organotvpic culture of fetal rat lung cells organized alveolar-like structures resembling those of in uiuo state were observed on the gelatin matrix. In contrast with in vivo state, fetal lung cells formed group of type ll pneumocytes predominently along the contours of the matrix. These cells have large apical surface, short microvilli and secreted materials which may be sunactant. These results suggested that an orsanotypic culture retaining epithelial- -mesenchvmal relationships is appropriate culture model to study the pulmonary epithelial cell (especially type ll pneumocvte) differentation.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1331-1337
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• 2006

A mathematical model has been proposed for the batch culture of Agaricus blazei with mixed carbon sources of glucose and dextrin. In the proposed model, the metabolism of A. blazei was divided into three parts: cell growth, exopolysaccharides (EPS) production, and another EPS production pathway activated by dextrin hydrolysis. Each pathway was described mathematically and incorporated into the mechanistic model structure. Batch cultures were carried out with six different carbon source compositions. Although parameters were estimated by using the experimental data from the two extreme cases such as glucose only and dextrin only, the model represented well the profiles of glucose, cell mass, and EPS concentrations for all the six different carbon source mixtures, showing a good interpolation capability. Of note, the lag in EPS production could be quite precisely predicted by assuming that the glucose-to-cell mass ratio was the governing factor for EPS production.

• KSBB Journal
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• 제14권5호
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• pp.534-538
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• 1999

식물세포배양을 위한 bioreactor의 운전조건 최적화를 위해 Nicotiana tabacum 현탁세포를 model system으로 bioreator의 종류, 교반기의 형태, 그리고 통기량에 따른 세포생장을 관찰하였다. Bioreactor로는 stirred tank bioreactor과 airlift bioreactor를 사용하였으며 두가지 배양기 모두 flask에서의 생장보다 낮은 생장을 보였으며 stirred tank bioreactor보다는 airlift bioreator에서 높은 세포농도를 얻을 수 있었다. 교반기의 종류에 따른 세포의 생장은 큰 차이가 없었으나 hollowed paddle impeller를 사용하였을 경우에는 배양기간 동안 세포의 크기가 작게 유지되었다. 통기량을 0.30 vvm으로 유지하는 경우에 가장 좋은 세포생장을 관찰할 수 있었으며 1.0 vvm이상의 통기량에서는 과도한 foam의 형성과 세포의 갈색화 현상을 보이며 세포의 생장도 저해되었다. 또한 통기량이 증가할수록 세포크기지수가 감소하는 결과를 보였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.84-91
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• 2000

Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

• Journal of Applied Biological Chemistry
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• 제48권4호
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• pp.170-177
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• 2005

Oriental medicinal herb extracts (OHE) showing anticancer activities were investigated for effectiveness as antiviral drugs. Infection of MuLV to cell line resulted in formation of giant syncytia. Number of giant syncytia in culture treated with OHE decreased by 40% compared to that of non-OHE-treated cell culture. To determine OHE effects on progeny release, RT-PCR was performed. In vivo animal studies demonstrated effectiveness of OHE as antiviral drug when administered orally. After OHE administration, viral cytopathic effects decreased. Infected mice showed splenomegaly and over-proliferation of lymphocytes with decreased CD4+ cell counts. These symptoms decreased in OHE-treated mice, indicating OHE maybe useful therapeutics against MuLV/MAIDS as Human Immunodeficiency Virus (HIV)/AIDS animal model. Results show XC plaque assay and in vivo MAIDS model using MuLV are suitable tools for screening anti-retroviral drug candidates.

• 한국수산과학회지
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• 제15권3호
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• pp.179-184
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• 1982

Lartobacillus bulgricus를 glucose를 제한기질로 하는 합성배지에서 연속배양한 결과를 요약하면 다음과 같다. 1. 본 실험 model은 Monod의 chemostat 이론을 적용시킬 수 있다. 2. 본 실험 model에서의 최대 cell production rate는 $0.178 g/1{\cdot}hr$로서 회석율 $0.414hr^{-1}$ 일 때이다. 3. 연속배양 결과 saturation constant($K_s$)는 7.69g/l, 최대비증식속도$(V_{max})$는 $0.62hr^{-1}$ 이었다. 4. wash out 현상은 $0.51hr^{-1}$에서 일어났으며cell yield coefficient는 0.016g/l 이었다.

• Journal of Preventive Veterinary Medicine
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• 제43권4호
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• pp.214-220
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• 2019

Although canine brucellosis has been known to be an important re-emerging zoonosis, the pathophysiological mechanisms of Brucella canis infection remains clues to be solved. Different culture models, single and co-culture models, were constructed with canine epithelial cells, D17 and macrophage, DH82 to investigate the induction of immune responses in in vivo B. canis infection. Expression of genes related with induction of immune responses, Th1, Th2 and Th17, was compared in the two different models after the bacterial infection. In this study, expression of cytokine genes, IL-1β, IL-5, IL-6, IL-10, IL-23, and TNF-α was quantified in the DH82 at different time points using RT-qPCR in the two different culture systems after the infection. Cytokine genes related with Th1, IL-1β and TNF-α and Th17, IL-6 and IL-23 were expressed with time-dependent manners in the both systems (p<0.05). However, increase of Th2-related cytokine genes expression was not detectable in the both systems by comparison with control. The expression of Th1 and Th17 related cytokine genes was earlier in single cell culture than those in co-culture model (p<0.05). In general, amounts of the expressed genes were shown higher in single cell model than those in co-culture models. This study indicate that Th1 and Th17-associated immune responses are central to B. canis infection in dogs. In addition, it suggests a specific role of epithelial cells in the B. canis infection in vivo, which should resolved in the further study.

• Journal of Pharmaceutical Investigation
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• 제34권5호
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• pp.393-399
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• 2004

For the efficient determination of activity against solid tumors, an in vitro tumor model that resembles the condition of in vivo solid tumors, is required. The purpose of this study was to establish a rapid culture method and viability assay for an in vitro 3-dimensional tumor model, multicellular spheroid (MCS). Among 12 human cancer cell lines, a few cell lines including DLD-1 (human colorectal carcinoma cells) formed fully compact MCS which was adequate for in vitro viability assay. DLD-1 MCS showed steady growth reaching $700\;{\mu}m$ diameter after 11 day culture. DLD-1 cells grown as MCS showed significant increase in $G_0/G_1$ phase compared to the monolayer cells (73.9% vs 45.7%), but necrotic regions or apoptotic cells were not observed. The cells cultured as MCS showed resistance to 5-FU (10.3 fold higher $IC_{50}$) compared to monolayers, however, tirapazamine (a hypotoxin) showed similar activity in both culture systems. In summary, MCS may be a valid in vitro model for activity screening of anticancer agents against human solid tumors and also exploitable for studying molecular markers of drug resistance in human solid tumors.

• The Korean Journal of Physiology and Pharmacology
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• 제1권3호
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• pp.285-296
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• 1997

Injury to brain transforms resting astrocytes to their reactive form, the hallmark of which is an increase in glial fibrillary acidic protein (GFAP), the major intermediate filament protein of their cell type. The overall glial response after brain injury is referred to as reactive gliosis. Glial-neuronal interaction is important for neuronal migration, neurite outgrowth and axonal guidance during ontogenic development. Although much attention has been given to glial regulation of neuronal development and regeneration, evidences also suggest a neuronal influence on glial cell differentiation, maturation and function. The aim of the present study was to analyze the effects of glial-hippocampal neuronal co-culture on GFAP expression in the co-cultured astrocytes. The following antibodies were used for double immunostaining chemistry; mouse monoclonal antibodies for confirm neuronal cells, rabbit anti GFAP antibodies for confirm astrocytes. Primary cultured astrocytes showed the typical flat polygonal morphology in culture and expressed strong GFAP and vimentin. Co-cultured hippocampal neurons on astrocytes had phase bright cell body and well branched neurites. About half of co-cultured astrocytes expressed negative or weak GFAP and vimentin. After 2 hour glutamate (0.5 mM) exposure of glial-neuronal co-culture, neuronal cells lost their neurites and most of astrocytes expressed strong CFAE and vimentin. In Western blot analysis, total GFAP and vimentin contents in co-cultured astrocytes were lower than those of primary cultured astrocytes. After glutamate exposure of glial-neuronal co-culture, GFAP and vimentin contents in astrocytes were increased to the level of primary cultured astrocytes. These results suggest that neuronal cell decrease GFAP expression in co-cultured astrocytes and hippocampal neuronal-glial co-culture can be used as a reactive gliosis model in vitro for studying GFAP expression of astrocytes.