Human <scp>BJ</scp> Fibroblasts is an Alternative to Mouse <scp>BALB</scp>/c 3T3 Cells in <i>In Vitro</i> Neutral Red Uptake Assay

Mannerström, Marika; Toimela, Tarja; Sarkanen, Jertta-Riina; Heinonen, Tuula

doi:10.1111/bcpt.12790

Cited by 36 publications

(20 citation statements)

References 22 publications

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“…It is important to note that the responses of HepG2, Caco‐2, A549 and HMEC‐1 to each chemical did not differ significantly from each other (Figure ). Our findings are in line with those of a previous report, which found that half the tested chemicals (13 out of 26) were more toxic to mouse BALB/c 3T3 cells than to human BJ fibroblasts; the remaining tested chemicals were equally (7 out of 26) or less toxic (6 out of 26) to 3T3 than BJ fibroblasts . These results are also in accordance with observations in MTT assay indicating that the 3T3 cell line was more sensitive than BJ human fibroblasts to various nanoparticles: the IC 50 values obtained for each type of nanoparticles were roughly 1.5‐fold lower in the experiments with the 3T3 mouse fibroblasts than with the BJ human fibroblasts .…”

Section: Discussionsupporting

confidence: 92%

“…For example, Knels et al recently reported the beneficial effects of infrared A on 3T3 fibroblast culture, which was regarded as a well‐defined cell line with a standard, stable growth rate . An additional argument given in favour of using the 3T3 cell line is that it is a globally accepted cell line for use in the neutral red uptake assay (NRU), which is a standardized method for acute toxicity testing in rodents . Another reason for using the 3T3 fibroblasts by some researchers is their good responsiveness to external stimuli; however, this feature has been primarily ascribed to cardiac fibroblasts: the main cellular constituents of the heart .…”

Section: Discussionmentioning

confidence: 99%

“…The capability of fibroblasts to respond to external stimuli together with their role in vivo (wound healing, angiogenesis and tissue regeneration) makes them an attractive resource for various biological studies . In fact, the mouse embryonic BALB/c 3T3 fibroblasts are the most common cell line used in the neutral red uptake (NRU) assay, which is a standardized test for estimating the starting dose for acute oral systemic toxicity tests in rodents . Likewise, the carcinogenic potential of chemicals is tested in the in vitro cell transformation assay (CTA) using BALB/c 3T3 cells .…”

Section: Introductionmentioning

confidence: 99%

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Differentiated mitochondrial function in mouse 3T3 fibroblasts and human epithelial or endothelial cells in response to chemical exposure

Boncler

Łukasiak²,

Dastych

et al. 2018

Basic Clin Pharma Tox

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Mouse 3T3 fibroblasts are commonly used for in vitro toxicity testing; however, their sensitivity to stimuli is not well defined. To assess the sensitivity of the 3T3 cell line, the study compared the changes in mitochondrial membrane potential (MMP) occurring after exposure to eight chemicals known to demonstrate pro-apoptotic activity (glycerol, isopropanol, ethanol, paracetamol, propranolol, cobalt chloride, formaldehyde and atropine). Five cell lines were used as follows: mouse 3T3 fibroblasts, human epithelial cells (A549, Caco-2 and HepG2) and human endothelial cells (HMEC-1). Cell sensitivity was assessed based on the total area under and over the dose-response curves (AUOC) in relation to baselines. The 3T3 fibroblasts had the highest AUOC values and were the most sensitive to the action of all the examined chemicals, with the exception of formaldehyde. Significant changes in MMP between the 3T3 cell line and other cells were observed after cell treatment with atropine (A549, Caco-2 or HMEC-1 cells vs 3T3 cells, P < 0.05), propranolol (A549 vs 3T3 cells, P < 0.01; HepG2 vs 3T3 cells, P < 0.05), cobalt chloride (A549 cells vs 3T3 cells, P < 0.01) or ethanol (HMEC-1 vs 3T3, P < 0.05). Formaldehyde appeared the most toxic compound for Caco-2 cells (Caco-2 vs 3T3 cells, P < 0.05). The surface areas (AUOC) calculated for each other chemical and obtained for HepG2, Caco-2, A549 and HMEC-1 did not differ significantly between cell lines. We postulate that mouse 3T3 fibroblasts demonstrate significantly higher relative sensitivity to many agents with toxic potential.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Differentiated mitochondrial function in mouse 3T3 fibroblasts and human epithelial or endothelial cells in response to chemical exposure

Boncler

Łukasiak²,

Dastych

et al. 2018

Basic Clin Pharma Tox

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“…The solutions of each well were then transferred to 96-well plate and read at 450 nm by microplate reader [ 17 ].…”

Section: Methodsmentioning

confidence: 99%

Pomegranate Juice Extract Decreases Cisplatin Toxicity on Peripheral Blood Mononuclear Cells

et al. 2020

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Background: Lung cancer is one of the most prevalent cancers worldwide. Chemotherapy regimens, targeted against lung cancer, are considered an effective treatment; albeit with multiple fatal side effects. An alternative strategy, nowadays, is using natural products. Medicinal plants have been used, in combination with chemotherapy, to ameliorate side effects. Aims: This study aims to investigate the antitumor effect of pomegranate juice (Punica granatum) on human lung adenocarcinoma basal epithelial cells (A549), to check the effect, when combined with low dose cisplatin (CDDP), at different doses. We also have evaluated the potential protective effect of pomegranate on normal peripheral blood mononuclear cells (PBMC). Methods: Phytochemical screening of the extract was done using standard classical tests. Total phenolic and sugar contents were determined using the Folin–Ciocalteu and anthrone reagents, respectively. The antioxidant activity of pomegranate was estimated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The viability of A549 cells and PBMC was evaluated using the neutral red assay. Results: Our results demonstrated that Punica granatum or pomegranate juice (with different concentrations: 150, 300, 600 µg/mL) contained high levels of flavonoids, alkaloids, tanins, lignins, terpenoids, and phenols. The DPPH method showed that pomegranate juice had a strong antioxidant scavenging activity. Neutral red showed that combining pomegranate juice with low dose CDDP (8 µg/mL) decreased the cell viability of A549 cells, by 64%, compared to treatment with CDDP or pomegranate alone. When added to low dose CDDP, pomegranate increased the viability of normal PBMC cells by 46%. Conclusions: These results demonstrated that pomegranate could potentiate the anticancer effect of low dose CDDP on human lung adenocarcinoma cells (A549 cells) and could as well decrease its toxicity on PBMC.

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“…Further data of in vitro experiments are needed for the weight of evidence approach in risk assessment and licensing . New end‐point tests predicting the mechanism by which a genotoxic chemical is acting to damage DNA were presented and the importance of cytotoxicity discussed . Techniques measuring alterations in yH2AX and p53 activation can be used to characterize the mode of action at various concentrations .…”

mentioning

confidence: 99%

The European Environmental Mutagenesis and Genomics Society Annual Meeting, 14–18 August 2016, Copenhagen, Denmark

Holme

Frötschl

Knudsen

2017

Basic Clin Pharma Tox

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The meeting was organized by EEMGS and ECOPA (The European Consensus Platform of Alternatives). It consisted of 10 sessions and a poster exhibition of 53 posters, 10 of which were by young scientists receiving grants from EEMGS, ECOPA, the 3R centre in Denmark and GUM. Together, there were four key note presentations, 50 oral presentations and 16 oral flash poster presentations. The number of participants was 180, from 18 countries (

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Human BJ Fibroblasts is an Alternative to Mouse BALB/c 3T3 Cells in In Vitro Neutral Red Uptake Assay

Cited by 36 publications

References 22 publications

Differentiated mitochondrial function in mouse 3T3 fibroblasts and human epithelial or endothelial cells in response to chemical exposure

Differentiated mitochondrial function in mouse 3T3 fibroblasts and human epithelial or endothelial cells in response to chemical exposure

Pomegranate Juice Extract Decreases Cisplatin Toxicity on Peripheral Blood Mononuclear Cells

The European Environmental Mutagenesis and Genomics Society Annual Meeting, 14–18 August 2016, Copenhagen, Denmark

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