The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity

Crouch, S. P. M.; Kozlowski, Roland Z.; Slater, K.; Fletcher, J.

doi:10.1016/0022-1759(93)90011-u

Cited by 805 publications

(588 citation statements)

References 11 publications

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“…We also tested an ATP bioluminescence assay that measures the ATP content of a sample, which is an indicator for metabolically active cells. 18 A sample is taken daily and mixed with the assay reagent containing luciferase, and the light signal is measured in a luminometer. The luciferase signal is proportional to the ATP A single representative curve is shown for each of the four assays (Figure 5a, lower panels).…”

Section: Decrease In Percentage Of Gfp-expressing Cells Is Due To Outmentioning

confidence: 99%

A competitive cell growth assay for the detection of subtle effects of gene transduction on cell proliferation

et al. 2011

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RNA interference (RNAi) is a sequence-specific gene silencing mechanism with therapeutic potential against many human pathogens. To obtain a durable therapeutic effect, stable transduction of target cells with for instance a lentiviral vector that expresses a short hairpin (shRNA) inducer of the RNAi pathway is necessary. Apart from the intended therapeutic effect, this treatment can induce negative effects on cell proliferation via off-target effects. A careful evaluation of the transduced cells is required to develop a safe gene therapy approach. Stably transduced cells are usually selected by expression of the enhanced green fluorescent protein (GFP) marker. In this study we show that the mixed transduction culture, containing both transduced GFP þ and untransduced GFP À cells, can simply be passaged to score the GFP þ /GFP À ratio by longitudinal flow cytometric analysis as a measure of the negative impact of the RNAi treatment on the cellular proliferation rate. We show that this assay is sensitive, easy to use and internally controlled for assessing subtle effects on cell proliferation of lentiviral transduction and transgene expression.

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Section: Decrease In Percentage Of Gfp-expressing Cells Is Due To Outmentioning

confidence: 99%

A competitive cell growth assay for the detection of subtle effects of gene transduction on cell proliferation

et al. 2011

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“…The CellTiter-Glo luminescent cell viability assay is based on the quantitation of ATP present, which signals the presence of metabolically active cells or viable cells [29]. An equal volume of reconstituted CellTiter-Glo reagent was added to 50 ml of cell suspension.…”

Section: Atp Cell Viability Assaymentioning

confidence: 99%

The role of laser fluence in cell viability, proliferation, and membrane integrity of wounded human skin fibroblasts following helium‐neon laser irradiation

2006

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Background: In medicine, lasers have been used predominantly for applications, which are broadly termed low level laser therapy (LLLT), phototherapy or photobiomodulation. This study aimed to establish cellular responses to Helium-Neon (632.8 nm) laser irradiation using different laser fluences (0.5, 2.5, 5, 10, and 16 J/cm 2 ) with a single exposure on 2 consecutive days on normal and wounded human skin fibroblasts. Materials and Methods: Changes in normal and wounded fibroblast cell morphology were evaluated by light microscopy. Changes following laser irradiation were evaluated by assessing the mitochondrial activity using adenosine triphosphate (ATP) luminescence, cell proliferation using neutral red and an alkaline phosphatase (ALP) activity assay, membrane integrity using lactate dehydrogenase (LDH), and percentage cytotoxicity and DNA damage using the Comet assay. Results: Morphologically, wounded cells exposed to 5 J/ cm 2 migrate rapidly across the wound margin indicating a stimulatory or positive influence of phototherapy. A dose of 5 J/cm 2 has a stimulatory influence on wounded fibroblasts with an increase in cell proliferation and cell viability without adversely increasing the amount of cellular and molecular damage. Higher doses (10 and 16 J/cm 2 ) were characterized by a decrease in cell viability and cell proliferation with a significant amount of damage to the cell membrane and DNA. Conclusions: Results show that 5 J/cm 2 stimulates mitochondrial activity, which leads to normalization of cell function and ultimately stimulates cell proliferation and migration of wounded fibroblasts to accelerate wound closure. Laser irradiation can modify cellular processes in a dose or fluence (J/cm 2 ) dependent manner. Lasers Surg.

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“…One Gray (Gy) is a measure of absorbed dose of radiation in the international system (SI) of units, and is defined as the amount of energy absorbed by a kilogram of matter in J kg -1 ; for an irradiator delivering a dosage of 1 Gy · min -1 , we exposed samples for either 2 or 16 mins to receive dosages of 2 Gy or 16 Gy, respectively. We measured the metabolic activity of the cells six days post-irradiation using the CellTiter-Glo ® (CTG) assay, which quantifies the level of ATP present in cells [53]. The sensitivity of cells to radiation (Eq 1 and 2) decreased with increasing seeding density of cells ( Fig.…”

Section: Sensitivity To Radiation Decreases With Increasing Cellular mentioning

confidence: 99%

Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system

et al. 2016

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(100-200 word limit)This work demonstrates the application of a 3D culture system, which is known as Cells-in-Gels-in-Paper or CiGiP, in evaluating the response of lung cancer cells to ionizing radiation. This system has four attributes: (i) multiple layers of paper, containing cell-embedded hydrogels, are assembled into a stack to form a thick (~800 µm) tissue-like construct, (ii) the metabolism of the cells, coupled with the boundary conditions imposed by an impermeable holder, generate a gradient of oxygen and nutrients that decreases monotonically in the stack, (iii) the construct has no peripheral components (e.g., pumps, tubing), that fits easily in an irradiator, and (iv) the construct can be disassembled into individual layers, allowing for the quantification of cellular phenotypes based on their position within the stack. With increased distance from the source of oxygenated media, cells show increased levels of hypoxia-inducible factor, decreased proliferation, and reduced sensitivity to ionizing radiation. The multi-layer culture setup for CiGiP also distinguished differences in the radiosensitivity of three isogenic variants of A549 cancer cells, which have known differences in their metastatic behavior in vivo. The CiGiP system can, therefore, capture aspects of radiosensitivity of populations of cancer cells related to mass-transport phenomenon inaccessible in traditional culture systems.

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The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity

Cited by 805 publications

References 11 publications

A competitive cell growth assay for the detection of subtle effects of gene transduction on cell proliferation

A competitive cell growth assay for the detection of subtle effects of gene transduction on cell proliferation

The role of laser fluence in cell viability, proliferation, and membrane integrity of wounded human skin fibroblasts following helium‐neon laser irradiation

Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system

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