• Preventive Nutrition and Food Science
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• v.8 no.1
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• pp.36-39
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• 2003

Many defined cell culture media were formulated over 3() years ago and may be deficient in certain micronutrients whose essentiality has only subsequently been recognised. The objective of this study was to evaluate whether alpha-minimal essential medium (MEM) supplemented with 10% foetal bovine serum contained sufficient selenium for optimal activity of the selenium containing enzymes cytosolic glutathione peroxidase (cGPx) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) in cultured Chinese hamster ovary (CHO) cells. Additionally, the effect of zinc deficiency and metallothionein (MT) overexpression on cGPx and PHGPx activity was studied. The addition of 100 nM of selenous acid to the culture medium increased cGPx expression by 10-fold and PHGPx by about 2-fold in both wild-type CHO-K1 cells and CHO-K1 cells overexpressing mouse MT-1. Zinc deficiency had no significant effect on enzyme activity, but cells overexpressing mouse MT-1 had higher levels of cGPx activity. Zinc deficiency decreased cell survival but overexpression of MT-1 was partially protective, probably because its presence in quantity favoured the uptake, sequestration and cellular retention of any remaining zinc. This study demonstrates that selenium in complete alpha-MEM is insufficient for optimal cGPx and PHGPx activity and may compromise the cellular response to oxidative stress.

• Bulletin of the Korean Chemical Society
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• v.25 no.9
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• pp.1303-1304
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• 2004

• 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.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.309-312
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• 2002

Exogenous glutamine synthetase (GS) was used efficiently as a selectable marker to identify successful transfectants for the production of erythropoietin (EPO) in Chines hamster ovary (CHO-K1) cells in the absence of glutamine. Inclusion of methionine sulphoximine (MSX), an inhibitor of glutamine synthetase, enabled further selection of clones with relatively high levels of transfected glutamine synthetase and EPO genes which were coupled together. In this study, a new cell line was established by using GS system and enhancement of EPO production by zinc ion was evaluated using the transfected CHO-K1 cell line under normal condition. It was found that EPO production from CHO-K1 cells was enhanced 40% when the optimal amount of zinc ion was added.

• Environmental Mutagens and Carcinogens
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• v.9 no.1
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• pp.13-22
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• 1989

The effect of novobiocin (NOV), and inhibitor of topoisomerase II, on ethyl methanesulfonate (EMS)-or bleomycin (BLM)-induced DNA repair synthesis was examined during the cell cycle of Chinese hamster ovary (CHO)-K1 cells. Three assays were employed in this study: cell survival, alkaline elution and unscheduled DNA synthesis. EMS was effective at killing CHO cells in G1 phase, wheras BLM preferentially killed cells in G2 and S phases. EMS induced the much more amount of DNA damage in G1 phase, while BLM induced in G2 phase than the other phases. The both of pre- and post-treatment with BOV inhibitied EMS- or BLM-induced DNA repair synthesis in G1 and G2 phases, and pretreatment with NOV inhibited more effectively than the post-treated group. These results suggested that CHO cells exhibited a differential sensitivity to cell lethality and DNA damage in relation to cell cycle according to used chemical agents, and that DNA topoisomerase II participated in an initial stage of DNA repair.

• KSBB Journal
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• v.21 no.4
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• pp.306-311
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• 2006

Cell preservation is indispensable in animal cell culture process and should be established according to the cell characteristics. In this study, we experimented hypothermic preservation of CHO cells that is widely used in pharmaceutical industry to produce therapeutic proteins and established a stable method of preservation. The highest viability of CHO cells was obtained when the cells were preserved using rolling tube, which means the cells should be suspended to avoid the cell lumping during the preservation. Also, we obtained superior preservation result under the anaerobic condition. To evaluate the serum-free media as a preservation solution, we investigated cell growth after hypothermic preservation using serum-free media. High cell viability and normal cell growth was observed during 10 days using serum-free media. Moreover, we found that more effective preservation when ${\alpha}$-tocopherol and retinoic acid is added to media as an additive. In the case of 1 liter large scale hypothermic preservation using established protocol, cell viability and growth rate was obtained as good as small scale one. This study is considered to be helpful for hypothermic preservation of CHO cells and large scale hypothermic preservation may be available through the further studies.

• KSBB Journal
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• v.28 no.2
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• pp.86-91
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• 2013

To date, various strategies have been studied to increase specific productivity in Chinese hamster ovary (CHO) cell cultures. Also, albumin-fusion platform is being applied to other important bioactive peptides with short half-lives. Here, we investigated the effects of silkworm gland hydrolysate (SGH) on the production of albumin-erythropoietin (Alb-EPO) in transgenic CHO cells. The viable cell density of CHO cells was increased by 13% in the medium containing 1 mg/mL SGH higher than in the control medium without SGH. In addition, the production of Alb-EPO was also 1.26- fold enhanced by reducing the early apoptosis of CHO cells. In conclusion, SGH could be used as a useful supplement for the enhancement of recombinant protein production.

• Korean Journal of Veterinary Pathology
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• v.5 no.2
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• pp.79-80
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• 2001

A gingival mass was detected from a 1-year-old female Great Dane dog. After surgical removal, the lesions recurred in 2 weeks and died of septicemia. Characteristic histologic features were large numbers of multinucleated giant cells which were connected with capillary vessels. Neovascularization was prominent with mononuclear and polynuclear cell infiltration. Overall features of these lesions except for giant cell infiltration were similar to granuloma. From these results, a gingival mass excised from a dog was diagnosed to be a peripheral giant cell granuloma (PGCG). This is the first report of canine subcutaneous PGCG in Korea.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.904-910
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• 2009

The development of effective cellular imaging requires a specific labeling method for targeting, tracking, and monitoring cellular/molecular events in the living organism. For this purpose, we studied the cellular uptake of isocyanide-functionalized silver and gold nanoparticles by surface-enhanced Raman scattering (SERS). Inside a single mammalian cell, we could monitor the intracellular behavior of such nanoparticles by measuring the SERS spectra. The NC stretching band appeared clearly at ${\sim}2,100cm^{-1}$ in the well-isolated spectral region from many organic constituents between 300 and 1,700 or 2,800 and $3,600cm^{-1}$. The SERS marker band at ${\sim}2,100cm^{-1}$ could be used to judge the location of the isocyanide-functionalized nanoparticles inside the cell without much spectral interference from other cellular constituents. Our results demonstrate that isocyanide-modified silver or gold nanoparticle-based SERS may have high potential for monitoring and imaging the biological processes at the single cell level.

• Animal cells and systems
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• v.5 no.3
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• pp.253-262
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• 2001

Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1＊ cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1＊ cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.