• Bulletin of the Korean Chemical Society
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• v.34 no.3
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• pp.943-945
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• 2013

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.231.2-231.2
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• 2003

The efficient EPO (Erythropoietin) expression system in Chinese Hamster Ovary (CHO) cells was devised through the removal of ammonium ion accumulated in the media by introducing urea cycle enzymes. Previously, we developed C05 cell by transfecting the carbamoly phosphate synthase (CPS) and ornithine transcarbamoylase (OTC) into the EPO expressing CHO cell, IBE. (omitted)

• KSBB Journal
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• v.13 no.4
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• pp.404-410
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• 1998

Three types of adsorbents were developed by immobilizing synthetic zeolite, Philipsite-Gismonine, in alginate, cellulose acetate and dialysis membrane for the in situ removal of ammonium ion which inhibits growth and productivity of animal cells such as CHO cells producing tPA. Ammonium ion removal efficiency and cell growth promoting effect with various immobilized adsorbents were evaluated and the membrane type was selected as an optimal immobilized adsorbent. The experiments were then simulated by adding 8mM ammonium chloride and immobilized adsorbent in order to validate the removal effect under high density cell cultures. The results showed increase in maximum cell density by three times, in cell viability, and in tPA productivity by 40%. And it was found that the promoting effects were more significant in case of high ammonium ion concentration system. It was also found that the optimum addition time for immobilized adsorbents was 48 hr in the absence of ammonium chloride addition and 72 hr in the presence of ammonium chloride addition.

• Proceedings of the Korean Society of Rheology Conference
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• 2001.06a
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• pp.89-92
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• 2001

• KSBB Journal
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• v.21 no.2
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• pp.110-117
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• 2006

During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing serum with 10% DMSO. But, in case of bioprocess under the serum-free conditions, including cultivation of cell lines and producing of pharmaceuticals, the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the serum-free cryopreservation on the CHO cells. To improve the survival rates of the cryopreserved CHO cells in serum-free condition, first, the effects of permeable and non-permeable additives for substitute serum on cell viability were investigated. The combination of 10% DMSO and 0.03 M raffinose in MEM-${\alpha}$ without serum indicated 76% of cell viability. However, it did not reach the survival rates(more than 95%) of the conventional cryopreservation. In the second, to evaluate the cryopreservative ability of the serum-free medium(SFM) we compared viability of the CHO cells cryopreserved in the SFMs(Sigma C5467, C4726, and C1707, JBI SF486 and PF486), the cryoprotectant(Genenmed CAN-1000) and the MEM-${\alpha}$ with serum. All solution contained 10% DMSO. As a result of the comparison, cryopreserved cells in the SFMs showed over 95% of viability and appeared predominant viability better than cryoprotectant CAN-1000. Finally, we assessed the stability of the CHO cells in the long-term cryopreservation(LTC) using SFM. Every three months, the cryopreserved CHO cells were thawed to estimate the cell viability and the recovery rates. Then, real-time RT-PCR analyzed the inserted CHO DHFR gene. All results for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion, it could be seen that the LTC in the SFM can substitute for serum using methods in the bioprocess proceeded by CHO cells for more than 18 months.

• IMMUNE NETWORK
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• v.16 no.1
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• pp.1-12
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• 2016

Th2 cell immunity is required for host defense against helminths, but it is detrimental in allergic diseases in humans. Unlike Th1 cell and Th17 cell subsets, the mechanism by which dendritic cells modulate Th2 cell responses has been obscure, in part because of the inability of dendritic cells to provide IL-4, which is indispensable for Th2 cell lineage commitment. In this regard, immune cells other than dendritic cells, such as basophils and innate lymphoid cells, have been suggested as Th2 cell inducers. More recently, multiple independent researchers have shown that specialized subsets of dendritic cells mediate Th2 cell responses. This review will discuss the current understanding related to the regulation of Th2 cell responses by dendritic cells and other immune cells.

• Experimental and Molecular Medicine
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• v.50 no.11
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• pp.1.1-1.10
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• 2018

Bone marrow-derived mesenchymal stem cells (BMMSCs) are used extensively for cardiac repair and interact with immune cells in the damaged heart. Macrophages are known to be modulated by stem cells, and we hypothesized that priming macrophages with BMMSCs would enhance their therapeutic efficacy. Rat bone marrow-derived macrophages (BMDMs) were stimulated by lipopolysaccharide (LPS) with or without coculture with rat BMCs. In the LPS-stimulated BMDMs, induction of the inflammatory marker iNOS was attenuated, and the anti-inflammatory marker Arg1 was markedly upregulated by coculture with BMMSCs. Myocardial infarction (MI) was induced in rats. One group was injected with BMMSCs, and a second group was injected with MIX (a mixture of BMMSCs and BMDMs after coculture). The reduction in cardiac fibrosis was greater in the MIX group than in the BMC group. Cardiac function was improved in the BMMSC group and was substantially improved in the MIX group. Angiogenesis was better in the MIX group, and anti-inflammatory macrophages were more abundant in the MIX group than in the BMMSC group. In the BMMSCs, interferon regulatory factor 5 (IRF5) was exclusively induced by coculture with macrophages. IRF5 knockdown in BMMSCs failed to suppress inflammatory marker induction in the macrophages. In this study, we demonstrated the successful application of BMDMs primed with BMMSCs as an adjuvant to cell therapy for cardiac repair.

• 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.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.97-97
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• 2002

Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.12 no.5
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• pp.30-35
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• 2013

In order to increase cell efficiency in crystalline silicon solar cell, reduction of light reflection is one of the essential problem. Until now silicon wafer was textured by wet etching process which has random patterns along crystal orientation. In this study, high aspect ratio patterns are manufactured by nano imprint process and reflectance could be minimized under 1%. After that, screen printed solar cell was fabricated on the textured wafer and I-V characteristics was measured by solar simulator. Consequently cell efficiency of solar cell fabricated using the wafer textured by nano imprint process increased 1.15% than reference solar cell textured by wet etching. Internal quantum efficiency was increased in the range of IR wave length but decreased in the UV wavelength. In spite of improved result, optimization between nano imprinted pattern and solar cell process should be followed.