• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.293-296
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• 2002

Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

• KSBB Journal
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• v.31 no.4
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• pp.270-276
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• 2016

In glycoprotein, Terminal sialic acid residues of N-linked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of $CO_2$ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and $37^{\circ}C$ respectively and the $CO_2$ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.

• Korean Journal of Food Science and Technology
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• v.36 no.3
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• pp.514-517
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• 2004

O157:H7, Salmonella typhimurium, Listeria monocytogenes were treated with aqueous chlorine dioxide to elucidate effect of aqueous chlorine dioxide treatment on major food-borne pathogenic bacteria. Survival plot of E.coli O157:H7 at 5 ppm chlorine dioxide showed typical first-order rate. After 5 min of treatment, cell number decreased by 1.5 log cycle. Survival plot slope gave D value of 3.37 min. S. typhimurium and L. monocytogenes showed biphasic curve. Aqueous chlorine dioxide treatment on S. typhimurium and L. monocytogenes resulted in bactericidal effect for 5 min, and thereafter no effect was observed under experimental conditions of this study. These results suggest concentration of chlorine dioxide is more important than treatment time, and 5 ppm chlorine dioxide treatment is not sufficient for sanitizing fresh vegetables.

• Journal of Biomedical Engineering Research
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• v.39 no.1
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• pp.22-29
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• 2018

Intervertebral disc(IVD) mainly consists of Annulus fibrosus(AF) and Nucleus pulposus(NP), playing a role of distributing a mechanical load on vertebral body. IVD tissue engineering has been developed the methods to achieve anatomic morphology and restoration of biological function. The goal of present study is to identify the possibilities for creating a substitute of IVD the morphology and biological functions are the same as undamaged complete IVD. To fabricate the AF and NP combine biphasic IVD tissue, AF tissue scaffolds have been printed by 3D bio-printing system with natural biomaterials and NP tissues have been prepared by scaffold-free culture system. We evaluated whether the combined structure of 3D printed AF scaffold and scaffold-free NP tissue construct could support the architecture and cell functions as IVD tissue. 3D printed AF scaffolds were printed with 60 degree angle stripe patterned lamella structure(the inner-diameter is 5mm, outer-diameter is 10 mm and height is 3 mm). In the cytotoxicity test, the 3D printed AF scaffold showed good cell compatibility. The results of histological and immunohistochemical staining also showed the newly synthesized collagens and glycosaminoglycans, which are specific makers of AF tissue. And scaffold-free NP tissue actively synthesized glycosaminoglycans and type 2 collagen, which are the major components of NP tissue. When we combined two engineered tissues to realize the IVD, combined biphasic tissues showed a good integration between the two tissues. In conclusion, this study describes the fabrication of Engineered biphasic IVD tissue by using enable techniques of tissue engineering. This fabricated biphasic tissue would be used as a model system for the study of the native IVD tissue. In the future, it may have the potential to replace the damaged IVD in the future.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.80-104
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• 1996

The development of routine techniques for the isolation and in vitro maintenance of conducting airway epithelial cells in a differentiated state provides an ideal model to study the factors involved in the regulation of the expression of mucocilicary differentiation. Several key factors and conditions have been identified. These factors and conditions include the use of biphasic culture technique to achieve mucociliary differentiation and the use of such stimulators, the thickness of collagen gel substratum, the calcium level, and vitamin A, and such inhibitors, the growth factors EGF and insulin, and steroid hormones, for mucous cell differentiation. Using the defined culture medium, the life cycle of the mucous cell population in vitro was investigated. It was demonstrated that the majority of the mucous cell population in primary cultures is not involved in DNA replication. However, the mucous cell type is capable of self-renewal in culture and this reproduction is vitamin A dependent. furthermore, differentiation from non-mucous cell type to mucous cell type can be demonstrated by adding back a positive regulator such as vitamin A to the “starved” culture. Cell kinetics data suggest that vitamin A-dependent mucous cell differentiation in culture is a DNA replication-independent process and the process is inhibited by TGF-${\beta}$1.

• Journal of the Korean Society of Food Culture
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• v.37 no.5
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• pp.438-445
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• 2022

Laminaria japonica is a type of brown algae widely consumed in Asian countries and contains many essential nutrients and exhibits anti-obesity, antioxidant, and anti-inflammatory effects. In this study, the antioxidant and immunomodulatory effects of a Laminaria japonica water extract (LJE) were investigated using an in vitro model. Mean total polyphenol content of LJE was 2.16±0.11 ㎍ GAE/mg, and LJE dose-dependently inhibited ABTS radical activity but did not scavenge DPPH radicals. In addition, LJE enhanced nitric oxide (NO) production and upregulated the mRNA expressions of proinflammatory cytokines (i.e., tumor necrosis factor-α and interleukin-6) in RAW 264.7 cells. On the other hand, LJE inhibited NO production and downregulated proinflammatory cytokine mRNA levels in endotoxin-stimulated RAW 264.7 cells. Thus, our data show that LJE has moderate antioxidant activity and biphasic immunomodulatory effects on RAW 264.7 cells. In summary, the study indicates that LJE has potential therapeutic use as a novel biphasic immuno-modulator.

• Journal of Periodontal and Implant Science
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• v.36 no.4
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• pp.797-808
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• 2006

Objective : The purpose of this study was to evaluate the physicochemical properties and cytocompatibility of microporous, spherical biphasic calcium phosphate(BCP) ceramics with a 60/40 $hydroxyapatite/{\beta}$ -tricalcium phosphate weight ratio for application as a bone graft substitute. Materials and Methods : Microporous, spherical BCP granules(MGSB) were prepared and their basic characteristics were compared with commercially available BCP(MBCP; Biomatlante, France) and deproteinized bovine bone mineral(Bio-Oss; GBistlich-Pharma, Switzerland, BBP; Oscotec. Korea), Their physicochemical properties were evaluated by scanning electron microscopy, X-ray diffractometry, Fourier-transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometer, and Brunauer-Emmett-Teller method. Cell viability and proliferation of MC3T3-El cells on different graft materials were evaluated. Results : MGSB granules showed a chemical composition and crystallinity similar with those in MBCP, they showed surface structure characteristic of three dimensionally, well-interconnected micropores. The results of MTT assay showed increases in cell viablity with increasing incubation times. At 4d of incubation, MGSB, MBCP and BBP showed similar values in optical density, but Bio-Oss exhibited significantly lower optical density compared to other bone substitutes(p <0,05). MGSB showed significantly greater cell number compared to other bone substitutes at 3, 5, and 7d of incubation(p <0,05), which were similar with those in polystyrene culture plates. Conclusion: These results indicated the suitable physicochemical properties of MGSB granules for application as an effective bone graft substitute. which provided compatible environment for osteoblast cell growth. However, further detailed studies are needed to confirm its biological effects on bone formation in vivo.

• Toxicological Research
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• v.14 no.2
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• pp.163-169
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• 1998

Fluoride (F), over a narrow concentration range, increases bone formation. Aluminum (Ai) too is biphasic in its action on bone, being mitogenic at very low levels and inhibitory at higher levels. Both F and Al are present in finished drinking water where the chemical interaction of these two agents is well characterized. F and AI, given individually, accumulate preferentially in bone. In addition. in vivo studies have shown that F causes the co-accumulation of Al in bone. Thus, it was necessary to determine the interactive effect of these two agents on bone mitogenesis. Calvaria were obtained from neonatal CD-1 mice and cultured with various concentrations of F (0.05~19 ppm) as NaF, Al (2 ppb~2 ppm) as $AlCl_3$ , or F and Al for 3 days at $37^{\circ}C$ on a rotating roller drum. Alkaline phosphatase activity in calvaria and $\beta$-glucuronidase activity in culture medium were determined as a measures of bone turnover. Alkaline phosphatase activity in calvaria was significantly increased by F (0.05~2 ppm) treatment and $\beta$-glucuronidase activity was slightly increased in the culture medium of calvaria treated with 0.3 ppm Al. The combination of 19 ppm F and 0.3 ppm Al increased alkaline phosphatase activity in calvaria, but did not affect $\beta$-glucuronidase activity, suggesting the interactive effect of fluoride and aluminum on bone turnover.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.263-268
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• 1989

To maximize the performance of recombinant cell fermentation process through optimizing environmental conditions, the production of invertase from recombinant Saccharomyces cerebisiae Containing SUC2 gene was studied as a model. The recombinant cells showed biphasic growth on glucose. Since the promoter of the SUC2 is regulated by the concentration of glucose in the medium, expression of invertase by recombinant yeast began when the glucose concentration decreased in a range of 0.25-0.4 g/L during the batch culture. Plasmid segregation occured frequently during glucose fermentation, and infrequently during ethanol oxidation. A rapid appearance of invertase activity with glucose was observed under nonaerated condition, and the maximum specific invertase activity was about 1.5 times as high as under aerobic condition, In fed batch culture, when n low level of glucose was continuously supplied to the tormentor after the time of glucose depletion during growth phase, specific and total invertase activity increased about 1.7 and 2.9 fold, respectively, in a batch culture.

• Journal of Oral Medicine and Pain
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• v.25 no.1
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• pp.53-62
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• 2000

Bone marrow culture systems are widely used to differentiate osteoclast-like cells in vitro using several osteotropic hormones. In this study, we isolated and cultured the mouse bone marrow cells with or without some osteotropic hormones such as parathyroid hormone(PTH), prostaglandin $E_2(PGE_2)$ and $l,25(OH)_2-vitamin$ $D_3$(Vit. $D_3$). We confirmed the formation of osteoclast-like cells morphologically and functionally by the expression of tartrate-resistant acid phosphatase(TRAP) and by their capability to resorb dentin slices. We also studied the effects of transforming growth $factor-{\beta}(TGF-{\beta})$ and epidermal growth factor(EGF) on the Vit. $D_3-induced$ osteoclast-like cell formation. In control, a few multinucleated cells were formed whereas PTH and $PGE_2$ increased the number of multinucleated cells. PTH, $PGE_2$ and Vit. $D_3$ induced the formation of TRAP-positive multinucleated cells. After culture of mouse bone marrow cells on the dentin slices with or without osteotropic hormones, giant cells with diverse morphology were found on the dentin slices under the scanning electronmicroscopy. After removing the attached cells, resorption pits were identified on the dentin slices, and the shape of resorption pits was variable. EGF increased the osteoclast-like cell formation induced by Vit. $D_3$, however, $TGF-{\beta}$ showed biphasic effect, which at low concentration, increased and at high concentration, decreased the osteoclast-like cell formation induced by Vit. $D_3$.