Application of recombinant fluorescent mammalian cells as a toxicity biosensor

2002 ◽  
Vol 46 (3) ◽  
pp. 51-56 ◽  
Author(s):  
E.J. Kim ◽  
Y. Lee ◽  
J.E. Lee ◽  
M.B. Gu
Keyword(s):  
Bisphenol A ◽  
Cell Lines ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Environmental Toxicants ◽  
Bacterial Biosensor ◽  
Biosensor System ◽  
Γ Ray ◽  
Ovary Cell Line

With respect to developing a more sensitive biosensor, a recombinant fluorescent Chinese Hamster Ovary cell line was used for the monitoring of various toxicants. Both cell lines, EFC-500 and KFC-A10, were able to detect toxicants sensitively. They were characterized with mitomycin C and γ-ray as genotoxicants and bisphenol A, nonylphenol, ziram and methyl bromide as possible and known EDCs. When compared to each other, the response of KFC-A10 was generally more informative and sensitive. Compared to typical bacterial biosensor systems, these cell lines offered a sensitivity of 2- to 50-fold greater for the tested chemicals. Based on these results, the use of mammalian cells offers a sensitive biosensor system that is not only fast, cheap and reproducible but also capable of monitoring the endocrine-like characteristics of environmental toxicants.

scholarly journals Escherichia coli aspartate transcarbamylase: a novel marker for studies of gene amplification and expression in mammalian cells.

1986 ◽  
Vol 6 (9) ◽  
pp. 3050-3058 ◽  
Author(s):  
J C Ruiz ◽  
G M Wahl
Keyword(s):  
Escherichia Coli ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Expression Vectors ◽  
Aspartate Transcarbamylase ◽  
Multifunctional Protein ◽  
High Concentrations ◽  
Atcase Activity ◽  
Ovary Cell Line

Eucaryotic expression vectors containing the Escherichia coli pyrB gene (pyrB encodes the catalytic subunit of aspartate transcarbamylase [ATCase]) and the Tn5 phosphotransferase gene (G418 resistance module) were transfected into a mutant Chinese hamster ovary cell line possessing a CAD multifunctional protein lacking ATCase activity. G418-resistant transformants were isolated and analyzed for ATCase activity, the ability to complement the CAD ATCase defect, and the ability to resist high concentrations of the ATCase inhibitor N-(phosphonacetyl)-L-aspartate (PALA) by amplifying the donated pyrB gene sequences. We report that bacterial ATCase is expressed in these lines, that it complements the CAD ATCase defect in trans, and that its amplification engenders PALA resistance. In addition, we derived rapid and sensitive assay conditions which enable the determination of bacterial ATCase enzyme activity in the presence of mammalian ATCase.

Escherichia coli aspartate transcarbamylase: a novel marker for studies of gene amplification and expression in mammalian cells

1986 ◽  
Vol 6 (9) ◽  
pp. 3050-3058
Author(s):  
J C Ruiz ◽  
G M Wahl
Keyword(s):  
Escherichia Coli ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Expression Vectors ◽  
Aspartate Transcarbamylase ◽  
Multifunctional Protein ◽  
High Concentrations ◽  
Atcase Activity ◽  
Ovary Cell Line

Eucaryotic expression vectors containing the Escherichia coli pyrB gene (pyrB encodes the catalytic subunit of aspartate transcarbamylase [ATCase]) and the Tn5 phosphotransferase gene (G418 resistance module) were transfected into a mutant Chinese hamster ovary cell line possessing a CAD multifunctional protein lacking ATCase activity. G418-resistant transformants were isolated and analyzed for ATCase activity, the ability to complement the CAD ATCase defect, and the ability to resist high concentrations of the ATCase inhibitor N-(phosphonacetyl)-L-aspartate (PALA) by amplifying the donated pyrB gene sequences. We report that bacterial ATCase is expressed in these lines, that it complements the CAD ATCase defect in trans, and that its amplification engenders PALA resistance. In addition, we derived rapid and sensitive assay conditions which enable the determination of bacterial ATCase enzyme activity in the presence of mammalian ATCase.

Counteracting effects of urea and betaine in mammalian cells in culture

1990 ◽  
Vol 258 (1) ◽  
pp. R198-R204 ◽  
Author(s):  
P. H. Yancey ◽  
M. B. Burg
Keyword(s):  
Mammalian Cells ◽  
Mdck Cells ◽  
Chinese Hamster ◽  
Renal Medulla ◽  
Nacl Stress ◽  
Ovary Cell ◽  
Growth And Survival ◽  
Colony Forming Efficiency ◽  
Forming Efficiency ◽  
Ovary Cell Line

Urea and methylamines, such as betaine, are among the major organic osmotic effectors accumulated by organisms under hyperosmotic (high NaCl) stress; the mammalian renal medulla also accumulates such compounds in antidiuresis. Studies on isolated proteins show that urea generally destabilizes protein structure, whereas methylamines are generally stabilizers capable of offsetting the effects of urea. The counteracting-osmolytes hypothesis predicts that cells exposed to high urea concentrations require methylamines for optimal function. In this study, urea, betaine, and other solutes (NaCl, glycerol, sorbitol) were added to growth medium of cultured mammalian cells under conditions in which most solutes entered the cells. For two renal [Madin-Darby canine kidney (MDCK) and PAP-HT25] and one nonrenal (Chinese hamster ovary) cell line, urea (greater than 100 mM) or betaine (greater than 50-100 mM) alone inhibited cell growth and survival, measured as colony-forming efficiency. However, the addition of betaine (up to 120 mM) to media with urea (50-300 mM) greatly increased colony-forming efficiency above that with urea alone. A similar, although less marked effect, was seen on colony sizes with MDCK cells. These results support the counteracting-osmolytes hypothesis.

scholarly journals Chinese hamster ovary cell line DXB-11: chromosomal instability and karyotype heterogeneity

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Victoria I. Turilova ◽  
Tatyana S. Goryachaya ◽  
Tatiana K. Yakovleva
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Chinese Hamster Ovary Cell ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Chromosome 8 ◽  
Ovary Cell ◽  
Differential Staining ◽  
Nucleolar Organizer Regions ◽  
Karyotype Heterogeneity

Abstract Background Chinese hamster ovary cell lines, also known as CHO cells, represent a large family of related, yet quite different, cell lines which are metabolic mutants derived from the original cell line, CHO-ori. Dihydrofolate reductase-deficient DXB-11 cell line, one of the first CHO derivatives, serves as the host cell line for the production of therapeutic proteins. It is generally assumed that DXB-11 is identical to DUKX or CHO-DUK cell lines, but, to our knowledge, DXB-11 karyotype has not been described yet. Results Using differential staining approaches (G-, C-banding and Ag-staining), we presented DXB-11 karyotype and revealed that karyotypes of DXB-11 and CHO-DUK cells have a number of differences. Although the number of chromosomes is equal—20 in each cell line—DXB-11 has normal chromosomes of the 1st and 5th pairs as well as an intact chromosome 8. Besides, in DXB-11 line, chromosome der(Z9) includes the material of chromosomes X and 6, whereas in CHO-DUK it results from the translocation of chromosomes 1 and 6. Ag-positive nucleolar organizer regions were revealed in the long arms of chromosome del(4)(q11q12) and both chromosome 5 homologues, as well as in the short arms of chromosomes 8 and add(8)(q11). Only 19 from 112 (16.96%) DXB-11 cells display identical chromosome complement accepted as the main structural variant of karyotype. The karyotype heterogeneity of all the rest of cells (93, 83.04%) occurs due to clonal and nonclonal additional structural rearrangements of chromosomes. Estimation of the frequency of chromosome involvement in these rearrangements allowed us to reveal that chromosomes 9, der(X)t(X;3;4), del(2)(p21p23), del(2)(q11q22) /Z2, der(4) /Z7, add(6)(p11) /Z8 are the most stable, whereas mar2, probably der(10), is the most unstable chromosome. A comparative analysis of our own and literary data on CHO karyotypes allowed to designate conservative chromosomes, both normal and rearranged, that remain unchanged in different CHO cell lines, as well as variable chromosomes that determine the individuality of karyotypes of CHO derivatives. Conclusion DXB-11and CHO-DUK cell lines differ in karyotypes. The revealed differential instability of DXB-11 chromosomes is likely not incidental and results in karyotype heterogeneity of cell population.

Sodium Nitroprusside Induces Internalization of Muscarinic Receptors Stably Expressed in Chinese Hamster Ovary Cell Lines

2002 ◽  
Vol 65 (2) ◽  
pp. 943-946 ◽  
Author(s):  
Roberto Maggio ◽  
Pascaline Barbier ◽  
Andrea Toso ◽  
Davide Barletta ◽  
Giovanni U. Corsini
Keyword(s):  
Sodium Nitroprusside ◽  
Cell Lines ◽  
Chinese Hamster Ovary Cell ◽  
Muscarinic Receptors ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Ovary Cell

scholarly journals Assignment of the human dihydrofolate reductase gene to the q11----q22 region of chromosome 5.

1984 ◽  
Vol 4 (10) ◽  
pp. 2010-2016 ◽  
Author(s):  
V L Funanage ◽  
T T Myoda ◽  
P A Moses ◽  
H R Cowell
Keyword(s):  
Dihydrofolate Reductase ◽  
Hybrid Cell ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Chromosome 5 ◽  
Dhfr Gene ◽  
Reductase Gene ◽  
Ovary Cell Line ◽  
Biochemical Analyses ◽  
Human Dihydrofolate Reductase

Cells from a dihydrofolate reductase-deficient Chinese hamster ovary cell line were hybridized to human fetal skin fibroblast cells. Nineteen dihydrofolate reductase-positive hybrid clones were isolated and characterized. Cytogenetic and biochemical analyses of these clones have shown that the human dihydrofolate reductase (DHFR) gene is located on chromosome 5. Three of these hybrid cell lines contained different terminal deletions of chromosome 5. An analysis of the breakpoints of these deletions has demonstrated that the DHFR gene resides in the q11----q22 region.

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