• Title/Summary/Keyword: Chinese hamster ovary cells

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Penetration of HEp-2 and Chinese Hamster Ovary Epithelial Cells by Escherichia coli Harbouring the Invasion-Conferring Genomic Region from Salmonella typhimurium

  • Park, Jeong-Uck;Hwang, Sang-Gu;Moon, Ja-Young;Cho, Yoon-Kweon;Kim, Dong-Wan;Jeong, Yong-Kee;Rhee, Kwang-Ho
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.270-274
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    • 2000
  • Pathogenic Salmonella typhimurium can invade the intestinal epithelium and cause a wide range of diseases including gastroenteritis and bacteremia in human and animals. To identify the genes involved in the infection, the invasion determinant was obtained from S. typhimurium 82/6915 and was subcloned into pGEM-7Z. A subclone DHl (pSV6235) invaded HEp-2 and Chinese hamster ovary epithelial cells and contained a 4.4 kb fragment of S. typhimurium genomic region. Compared with the host strain E. coli DHl, the subclone DHl (pSV6235) invaded cultured HEp-2 and Chinese hamster ovary cells at least 75- and 68-fold higher, respectively. The invasion rate of E. coli DHl for the cells significantly increased by harbouring the genomic region derived from pathogenic S. typhimurium 82/6915.

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Correlation Between Enhancing Effect of Sodium Butyrate on Specific Productivity and mRNA Transcription Level in Recombinant Chinese Hamster Ovary Cells Producing Antibody

  • Jeon, Min-Kyoung;Lee, Gyun-Min
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.1036-1040
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    • 2007
  • Sodium butyrate (NaBu) has been used to enhance protein expression levels in mammalian cell culture. To determine the clonal variability of recombinant Chinese hamster ovary (rCHO) cells in response to NaBu addition regarding specific antibody productivity $(q_{Ab})$, three rCHO clones were subjected to different concentrations of NaBu. For all three clones, NaBu addition inhibited cell growth and decreased cell viability in a dose-dependent manner. On the other hand, the enhancing effect of NaBu on $q_{Ab}$ varied significantly among the clones. NaBu addition enhanced the antibody production of only one clone. RT-PCR analysis revealed that the changes in $q_{Ab}$ correlated linearly with those of the mRNA transcription level. Thus, it was concluded that the different enhancing effects of NaBu on protein expression in rCHO cell clones resulted from their different mRNA transcription levels.

Chromosomal Aberrations in Chinese Hamster Ovary Cells Induced by Kojic Acid (Kojic Acid에 의해 유기된 Chinese Hamster 난소세포의 염색체 변이)

  • Lee, Yang-Soon;Wei, Cheng-I
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.454-459
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    • 1992
  • Kojic acid, a fungal metabolite produced by some species of Aspergillus and Penicillium, was found to induce chromosomal aberrations in Chinese hamster ovary cells in the presence or absence of the rat liver homogenate (59 mix). All categories of chromosomal aberrations increased with increased doses of kojic acid. Based on the this result, kojic acid was assumed to be a kind of mutagens. On the potential toxicity of this compound it becomes evident that kojic acid would not be used as a food additive at this time.

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The efficient Erythropoietin expression system in Chinese Hamster Ovary cells by introduction of urea cycle enzymes

  • Lee, Yun-Jeong;Kim, Jung-Kwon;Kim, Hyung-Jin;Kim, Na-Young;Kim, Jung-Hoe;Kim, Hong-Jin
    • 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)

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A Study on the Induction of Sister-Chromatid Exchanges in Chinese Hamster Ovary $K_1$ Cells by Exposure to Cadmium (Chinese Hamster Ovary $K_1$ 세포의 자매염색분체교환에 미치는 카드뮴의 영향에 관한 연구)

  • Hwang, In-Kyung;Kim, Don-Kyoun
    • Journal of Preventive Medicine and Public Health
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    • v.23 no.2 s.30
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    • pp.178-184
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    • 1990
  • In order to examine the mutagenicity of cadmium dichloride the author studied the induction of sister chromatid exchanges in chinese hamster ovary $K_1$ cells which treated with cadmium dichloride at various concentrations. The results obtained were as follows : 1. In cells treated with $10^{-4}M$ cadmium dichloride, a small number of cells were viable but no mitosis was bound. 2. The frequencies of sister chromatid exchanges in cells treated with $10^{-5}M\;and\;10^{-6}M$ cadmium dichloride as $10.7{\pm}1.9\;and\;8.3{\pm}2.1$, respectively, were significantly increased for control ($6.0{\pm}2.3$). (P<0.05). 3. There were dose-dependent relationship between the concentration of cadmium dichloride and frequency of sister chromatid exchanges in cells treated with cadmium dichloride at concentration ranging from $10^{-5}\;to\;10^{-7}M$.

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Zinc and Selenium Requirements for Glutathione Peroxidase Activity and Cell Survival in Chinese Hamster Ovary Cells Overexpressing Metallothionein

  • Kwun, In-Sook;John R. Arthur;John H. Beattie
    • 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.

Effects of Silkworm Gland Hydrolysate on Albumin-erythropoietin Production in Transgenic Chinese Hamster Ovary Cells (형질전환 Chinese Hamster Ovary 세포에서 Albumin-erythropoietin의 생산시 Silkworm Gland Hydrolysate의 효과)

  • Choi, Min-Ho;Cha, Hyun-Myoung;Kim, Sun-Mi;Choi, Yong-Soo;Kim, Dong-Il
    • 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.

Penetration of HEp-2 and Chinese Hamster Ovary Epithelial Cells by Escherichia coli Harbouring the Invasion-Conferring Genomic Region from Salmonella typhimurium

  • 박정욱;황상구;문자영;조용권;김동완;정용기
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.270-270
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    • 2002
  • Pathogenic Salmonella typhimurium can invade the intestinal epithelium and cause a wide range of diseases including gastroenteritis and bacteremia in human and animals. To identify the genes involved in the infection, the invasion determinant was obtained from S. typhimurium 82/6915 and was subcloned into pGEM-7Z. A subclone DHl (pSV6235) invaded HEp-2 and Chinese hamster ovary epithelial cells and contained a 4.4 kb fragment of S. typhimurium genomic region. Compared with the host strain E. coli DHl, the subclone DHl (pSV6235) invaded cultured HEp-2 and Chinese hamster ovary cells at least 75- and 68-fold higher, respectively. The invasion rate of E. coli DHl for the cells significantly increased by harbouring the genomic region derived from pathogenic S. typhimurium 82/6915.

High-Level Expression of Recombinant Human Bone Morphogenetic Protein-4 in Chinese Hamster Ovary Cells

  • PARK JUNHO;YU SUNGRYUL;YOON JAESEUNG;BAEK KWANGHEE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1397-1401
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    • 2005
  • Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

Expression of Recombinant Human Follicle-stimulating Hormone in the Chinese Hamster Ovary Cell

  • Park, Ji-Hyun;Kim, Nam-Hyung;Hosup Shim;Kim, Teoan
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.100-100
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    • 2002
  • As an preliminary experiment for making transgenic animals producing human follicle stimulating hormone (hFSH), we tried to express recombinant hFSH gene in vitro. hFSH is a heterodimeric glycoprotein hormone produced in the anterior pituitary gland. The hormone is essential in the regulation of reproductive processes, such as follicular development and ovulation. Genes encoding the common gonadotrophin alpha subunit and FSH-specific beta subunit were inserted into retroviral vectors under the control of the rat beta actin promoter. Gene transfer to the Chinese hamster ovary (CHO) cells was done by infection of the retroviruses harvested from PT67 packaging cells transfected with recombinant retrovirus vector DNA. After selection with G4l8, PCR and RT-PCR analyses of the G4l8-resistant CHO cells showed successful transfer and expression of both ${\alpha}$ and ${\beta}$ fragments of the FSH gene.

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