• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.621-621
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• 2003

I. Objectives In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2(IL-2), Interleukin-4(IL-4) and Transforming growth $factor-{\beta}1(TGF-{\beta}1)$ in human lymphocytes. II. Materials and methods Enterococcus faecalis(ATCC29212) strains were used in this study. Strains were grown in 1-liter cultures in 85% N2-10% H2-5% $CO_2$chamber for 3 days at $37^{\circ}C$. The medium used was 3.7% brain heart infusion broth. Bacterial cells harvested from 1-liter cultures were washed, suspended in 20ml of phosphate-buffered saline(PBS). Suspensions of bacterial cells were disrupted by sonication on ice for 5 min. Protein concentration was determined by the Bicinchoninic acid(BCA) protein assay.(omitted)

• Archives of Plastic Surgery
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• 제34권4호
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• pp.426-431
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• 2007

Purpose: The aim of this study is to compare the effects of bone marrow stromal cells(BSCs) and fibroblasts on wound healing activity in vivo, especially on epithelization. Methods: The fibroblasts and BSCs were harvested from patients and cultured. Ten Spague-Dawley white rats were used. A 5 mm punches were made to excise skin and subcutaneous tissue in a round fashion at six sites on the back area of each rat. Four hundred thousand cells suspended in 0.05 ml fibrinogen were applied to the created wounds. The cells in group I, II, and III were no cells, fibroblasts and BSCs. The lengths of epithelial gap at the widest wound site were compared with autopsy specimens obtained on the 6th day after cell therapy under light microscope. Statistical comparisons were performed using the Mann-Whitney U-test, and the p value < 0.05 was considered statistically significant. Results: The best epithelization was also seen in the BSC group, followed by fibroblast and no cell groups.Conclusion: These results demonstrate that BSC has superior effect on stimulating wound healing than fibroblast, which is currently used for wound healing.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.997-1006
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• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• 생약학회지
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• 제28권4호
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• pp.225-232
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• 1997

300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.21-26
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• 2012

Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

• 한국미생물·생명공학회지
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• 제24권6호
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• pp.717-725
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• 1996

We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

• KSBB Journal
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• 제15권6호
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• pp.609-614
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• 2000

최근 항암제로서 관심을 끌고있는 camptothecin올 생산하기 위한 희수(Camptotheca acuminata) 현탁세포 배양과 cam­p ptothecin의 생산을 촉진하는 연구를 수행하였다. 장기간 배양 으로 발생되는 이차대사산물 생산능력의 변이가 관찰되어 이 를 극복하고자 세포의 응집을 통한 camptothecin의 생산을 촉진하고자 하였다. 세포의 응집은 hybrid 배지와 sucrose 4 %일 때에 가장 많이 유도되었고, 이때 camptothecin의 생산량도 $18.04{\times}10^{-4} mg/L$로 가장 높은 생산성을 가졌다. 또한, 세포의 응집을 유도하기 위한 일환으로 교반속도를 변화시켜 이에 대한 효과를 알아보았는데, 100rpm에서 가장 많은 세포응집이 유도되었으며, camptothecin의 생산 역시 $19.4{\times}10^{-4} mg/L$로 가장 높은 결과를 나타내었다.

• Archives of Pharmacal Research
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• 제22권2호
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• pp.119-123
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• 1999

Parathyroid hormone (PTH) treatment of bone and kidney-derived cells not only activates adenyly cyclase buy also increases intracellular free calcium, and translocates protein kinase C (PKC) from cytosol to plasma membranes. We have found that acute phorbol ester pretreatment significantly decreases PTH-induced calcium transients and the effect of phorbol ester was antagonized by staurosporine (ST). Although the major effect of ST in that study was the reversal of the action of phorbol ester, it appeared that ST may also have promoted the effect of PTH directly. To further investigate the observation, we examined the effect of ST on the intracellular calcium transients induced by PTH and $\alpha$-thrombin ($\alpha$-TH). For calcium transient experiments, UMR-106 cells were loaded with 2 mM fluo-acetoxymethylester for 30 min at room temperature. The cells were then washed and suspended in buffer containing 1 mM calcium. Fluorescence was detected at 530 nm, with excitation at 505 nm. ST alone did not cause calcium transients, but enhanced the transients elicited by PTH response. added 5 min before the hormone. Another protein kinase inhibitor H-7 likewise enhanced the calcium responses elicited by PTH, while genistein did not affect PTH response. Calcium transients elicited by $\alpha$-TH were also enhanced by ST. The results suggest that there might be tonically activated endogenous protein kinase(s) which inhibit calcium signaling of some calcemic agents.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.772-779
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• 2005

Ammonia-oxidizing bacteria (AOB) were enriched by repeating fed-batch cultivations in an AOB-selective medium of activated sludges from a domestic wastewater treatment plant. Enriched culture showed strong capabilities of ammonia oxidation [0.810 mg $NH_4^+$-N/mg mixed liquor suspended solids (MLSS)$\cdot$day] as well as $NO_x^-$-N production (0.617 mg $NO_x^-$-N/ mg MLSS$\cdot$day). Degree of enrichment was examined through fluorescent in situ hybridization (FISH) analyses using an AOB-specific Cy3-labeled oligonucleotide probe (NSOl90) and terminal-restriction fragment length polymorphism (T-RFLP) analyses. FISH analyses confirmed that the fraction of AOB among 4',6-diamidino-2-phenylindole (DAPI)-stained cells increased from about less than $0.001\%$ to approximately $42\%$ after enrichment of AOB, and T-RFLP analyses showed that bacterial community became simpler as enrichment was continued. When the enriched culture of AOB was added (150 mg/l as dry suspended solid) to the normal activated sludge (3,000 mg/l as dry suspended solid), nitrification efficiencies were improved from 0.020 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.041 mg $NO_x^-$-N/mg MLSS$\cdot$day in a synthetic wastewater and also from 0.0007 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.0918 mg $NO_x^-$-N/mg MLSS$\cdot$day in a real domestic wastewater. Therefore, it is expected that this enrichment method could be used for improving efficiency of nitrification in wastewater treatment plants.

• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.142-149
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• 1991

Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.