The frame multicentre project on In vitro cytotoxicology

Knox, P. P.; Uphill, P.F.; Fry, J. R.; Benford, J.; Balls, Michael

doi:10.1016/0278-6915(86)90092-x

Cited by 187 publications

(61 citation statements)

References 4 publications

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“…Kenacid blue assay determines the total cellular protein content by measuring the uptake of dye (Knox et al, 1986;Putnam et al, 2002;Memon and Pratten, 2009), which helps to assess the total cell number in comparison to the control (Putnam et al, 2002). The procedure is described in detail by Ahir and Pratten (2011).…”

Section: End Pointsmentioning

confidence: 99%

Evaluation of Bupropion Hydrochloride Developmental Cardiotoxic Effects in Chick Cardiomyocyte Micromass Culture and stem cell derived Cardiomyocyte Systems

Qureshi

Latif

Parker

et al. 2014

Birth Defects Research Pt B

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The use of antidepressant drug bupropion hydrochloride (BPN) during pregnancy results in increased cardiovascular anomalies. In this study, BPN developmental cardiotoxic effects in in vitro system were evaluated using chick cardiomyocyte micromass (MM) culture system and mouse embryonic stem cell derived cardiomyocyte (ESDC) system. In MM system, the cardiomyocyte contractile activity significantly decreased only at BPN 200 μM, while in ESDC system BPN concentration above 75 μM resulted in decreased contractile activity. The increase in drug concentration also affected the cardiomyocyte viability and total cellular protein content in both systems, but in ESDC system the cell viability failed to attain significant difference. The drug failed to induce reactive oxygen species production in both systems, but has affected the cardiac connexin43 expression especially in MM system. We observed that BPN showed developmental cardiotoxic effects irrespective of the stage of cardiac development in both in vitro systems.

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Section: End Pointsmentioning

confidence: 99%

Evaluation of Bupropion Hydrochloride Developmental Cardiotoxic Effects in Chick Cardiomyocyte Micromass Culture and stem cell derived Cardiomyocyte Systems

Qureshi

Latif

Parker

et al. 2014

Birth Defects Research Pt B

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“…This method is based on that of Knox et al 27 Briefly, after 90 min, the medium was discarded from the wells and the cultures washed twice with PBS. After addition of 150 L fixative to each well, the plate was gently agitated for 20 min.…”

Section: Kenacid Blue R Dye Binding Assaymentioning

confidence: 99%

Fibronectin interactions with osteoblasts: Identification of a non–integrin‐mediated binding mechanism using a real‐time fluorescence binding assay

Sim

Cladera

O’Shea

2003

J Biomedical Materials Res

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Fibronectin (Fn) is an extracellular matrix protein that interacts with specific integrins on the cell surface, initiating signal transduction processes that lead to a reorganization of the cytoskeleton and the assembly of focal adhesions. Cell surface proteoglycans or glycosaminoglycans (GAGs) such as heparan sulfate are also known to participate in the interaction of Fn with the cell surface by binding to two different heparin-binding domains. The influence of Fn and GAGs on the spreading and differentiation of human osteoblasts was also previously described. In the current work, a method developed in our laboratory is established to evaluate the interaction between Fn and human osteoblasts and the influence of GAGs on such interactions. This technique makes use of fluoresceinphosphatidylethanolamine (FPE) such that when inserted into the lipidic bilayer, it acts as a fluorescent indicator of membrane interactions. The results indicate that the binding profile of Fn with the osteoblast cell surface is best represented by a hyperbolic single binding site model with a membrane affinity of 120 nM. Removal of cell surface heparan sulfate by treatment with heparitinase indicates that the cell surface moiety is directly involved in the binding process. Studies directed to assess the influence of heparin on the interaction of Fn with osteoblasts reveal that although it does not hamper Fn binding to the cell surface, it blocks the initial attachment to Fn-coated surfaces, indicating that binding to the integrin receptor alone is not enough to promote cell attachment but that the participation of the cell-surface GAGs is also a necessary condition.

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“…The method used was that proposed by Knox et al [9], in which kenacid blue is employed to stain cell protein. The absorbance of the solution in each well was measured at 620 nm with a Microplate Reader (EL 312 c).…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

Cytotoxicity and mutagenicity of 6-aminonicotinamide in Chinese hamster ovary cells

Pozuelo

Maria

1996

Teratog. Carcinog. Mutagen.

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A strong teratogen—6‐aminonicotinamide (6‐AN)—was tested for its ability to induce cytotoxicity and mutagenicity in Chinese hamster ovary (CHO) cells. Tests were performed in the presence and absence of a metabolic activation system (S‐9 mix). Cytotoxicity was evaluated in CHO cells by the total protein content. The two single‐gene mutation systems in CHO cells have been investigated. Both involve evaluating the response of the hypoxanthine‐guanine phosphoribosyl transferase (HGPRT) locus and specific inhibitors of the cellular (Na+ + K+)‐ATPase using 6‐thioguanine and ouabain as selective agents, respectively. From our results, 6‐AN showed a higher cytotoxic effect at concentrations over 1 × 10−1 mg/ml. Cytotoxicity was significantly different with and without S‐9 mix. 6‐AN was cytotoxic per se, however, when 6‐AN was biotransformed, in the presence of S‐9 mix no biological activity (cytotoxic) was detected. Non‐significant mutagenic activity was detected with 6‐AN in the presence and in the absence of the metabolic activation system. © 1996 Wiley‐Liss, Inc.

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The frame multicentre project on In vitro cytotoxicology

Cited by 187 publications

References 4 publications

Evaluation of Bupropion Hydrochloride Developmental Cardiotoxic Effects in Chick Cardiomyocyte Micromass Culture and stem cell derived Cardiomyocyte Systems

Evaluation of Bupropion Hydrochloride Developmental Cardiotoxic Effects in Chick Cardiomyocyte Micromass Culture and stem cell derived Cardiomyocyte Systems

Fibronectin interactions with osteoblasts: Identification of a non–integrin‐mediated binding mechanism using a real‐time fluorescence binding assay

Cytotoxicity and mutagenicity of 6-aminonicotinamide in Chinese hamster ovary cells

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