A new, rapid colorimetric assay for quantitative determination of cellular proliferation, growth inhibition, and viability

Rieck, Peter; Peters, Dirk; Hartmann, Christian; Courtois, Yves

doi:10.1007/bf02387288

Cited by 22 publications

(9 citation statements)

References 10 publications

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“…Cells were seeded in 24-well culture plates at 10 ~ cells/well and proliferation was quantified using the supravital dye Janus green (26). DNA was assayed with bisbenzimide (18) as modified by Bonis (7).…”

Section: Cell Proliferation and Dna Assaysmentioning

confidence: 99%

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Characterization of human osteoblastic cells: Influence of the culture conditions

Rattner¹,

Sabido²,

Massoubre³

et al. 1997

In Vitro Cell.Dev.Biol.-Animal

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Human osteoblastic cells were isolated enzymatically from adult human spongy bone and grown in MEM-Ham F12 1:1 medium supplemented with 2% Ultroser (USM). They were subcultured and examined for osteoblast features by morphological, histological, and biochemical approaches. The cells had a characteristic polyhedral morphology and produced a high level of alkaline phosphatase (ALKP). Confluent cultures were uniformly stained for ALKP and flow cytometry analysis with fluorescein diphosphate gave a single peak signal, reflecting a highly positive population, distinct from cultures of fibroblasts. The ALKP activity was stimulated by 1,25 (OH)2 vitamin D3. CD 44 was strongly expressed in these cultures, although osteoblasts are negative in vivo and osteocytes are positive. The main collagen synthesized was type I collagen and osteocalcin was produced after stimulation by vitamin D3. 10 mM betaGP induced mineralization and microprobe analysis of the crystals showed a composition close to hydroxyapatite. Changing the culture conditions to MEM-10% calf serum acted on cell behavior: it reduced the production of these biochemical markers of osteoblasts and the morphology became fibroblastlike with more rapid cell multiplication. The parameter most affected by the change in culture medium was ALKP, which was selected as the determinant criterion for defining an osteoblast culture. ALKP activity was then used to characterize a culture of cells seeded in a collagen gel.

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Section: Cell Proliferation and Dna Assaysmentioning

confidence: 99%

“…Many culture systems have been developed using osteoblastlike cells of animal or human origin, derived from normal bone or osteosarcomas (2,5,13,24,26,27,29,31). But the use of animal cell lines raises the problem of extrapolation to humans.…”

Section: Introductionmentioning

confidence: 99%

Characterization of human osteoblastic cells: Influence of the culture conditions

Rattner¹,

Sabido²,

Massoubre³

et al. 1997

In Vitro Cell.Dev.Biol.-Animal

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“…Following chemical exposures, cytotoxicity was assessed by examining cellular proliferation and viability. JG dye was used to assess cellular proliferation . CFDA was used to assess viability.…”

Section: Methodsmentioning

confidence: 99%

“…JG dye was used to assess cellular proliferation. (22) CFDA was used to assess viability. After 24-hour chemical exposures, assays were performed.…”

Section: Cytotoxicitymentioning

confidence: 99%

Binary Mixtures of Polycyclic Aromatic Hydrocarbons Display Nonadditive Mixture Interactions in an In Vitro Liver Cell Model

Gaskill

Bruce

2015

Risk Analysis

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Polycyclic aromatic hydrocarbons (PAHs) have been labeled contaminants of concern due to their carcinogenic potential, insufficient toxicological data, environmental ubiquity, and inconsistencies in the composition of environmental mixtures. The Environmental Protection Agency is reevaluating current methods for assessing the toxicity of PAHs, including the assumption of toxic additivity in mixtures. This study was aimed at testing mixture interactions through in vitro cell culture experimentation, and modeling the toxicity using quantitative structure-activity relationships (QSAR). Clone-9 rat liver cells were used to analyze cellular proliferation, viability, and genotoxicity of 15 PAHs in single doses and binary mixtures. Tests revealed that many mixtures have nonadditive toxicity, but display varying mixture effects depending on the mixture composition. Many mixtures displayed antagonism, similar to other published studies. QSARs were then developed using the genetic function approximation algorithm to predict toxic activity both in single PAH congeners and in binary mixtures. Effective concentrations inhibiting 50% of the cell populations were modeled, with R(2) = 0.90, 0.99, and 0.84, respectively. The QSAR mixture algorithms were then adjusted to account for the observed mixture interactions as well as the mixture composition (ratios) to assess the feasibility of QSARs for mixtures. Based on these results, toxic addition is improbable and therefore environmental PAH mixtures are likely to see nonadditive responses when complex interactions occur between components. Furthermore, QSAR may be a useful tool to help bridge these data gaps surrounding the assessment of human health risks that are associated with PAH exposures.

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“…Briefly to measure the protein content, the cells were harvested from the culture plate and suspended in lysis buffer, using a procedure described by Zhang [216]. The appropriate kits were used following manufacturer's protocols.…”

Section: Monitoring Of No Ros Sos Mmp and Ldh Level(s)mentioning

confidence: 99%

Nanosafety

Bashir

Liu

2015

Advanced Nanomaterials and Their Applications in Renewable Energy

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A new, rapid colorimetric assay for quantitative determination of cellular proliferation, growth inhibition, and viability

Cited by 22 publications

References 10 publications

Characterization of human osteoblastic cells: Influence of the culture conditions

Characterization of human osteoblastic cells: Influence of the culture conditions

Binary Mixtures of Polycyclic Aromatic Hydrocarbons Display Nonadditive Mixture Interactions in an In Vitro Liver Cell Model

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