Comparison of XTT and Alamar blue assays in the assessment of the viability of various human cancer cell lines by AT-101 (−/− gossypol)

Uzunoğlu, Selim; Karaca, Burçak; Atmaca, Harika; Kısım, Aslı; Sezgin, Canfeza; Karabulut, Bülent; Uslu, Rüçhan

doi:10.3109/15376516.2010.508080

Cited by 38 publications

(31 citation statements)

References 14 publications

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“…[58,59], Trichomonas vaginalis [60], Trypanosoma brucei [61], Leishmania sp. [36], Aspergillus fumigatus [62], as well as cancer [63,64] and Clostridium perfringens e-toxin [65]. The resazurin reduction assay assesses cell viability by measuring the The resulting output shows each individual nucleus with distinct color (A color map is drawn in frame C).…”

Section: Discussionmentioning

confidence: 99%

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

et al. 2013

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Chikungunya virus (CHIKV) is a mosquito-borne arthrogenic alphavirus that causes acute febrile illness in humans accompanied by joint pains and in many cases, persistent arthralgia lasting weeks to years. The re-emergence of CHIKV has resulted in numerous outbreaks in the eastern hemisphere, and threatens to expand in the foreseeable future. Unfortunately, no effective treatment is currently available. The present study reports the use of resazurin in a cell-based high-throughput assay, and an image-based high-content assay to identify and characterize inhibitors of CHIKV-infection in vitro. CHIKV is a highly cytopathic virus that rapidly kills infected cells. Thus, cell viability of HuH-7 cells infected with CHIKV in the presence of compounds was determined by measuring metabolic reduction of resazurin to identify inhibitors of CHIKV-associated cell death. A kinase inhibitor library of 4,000 compounds was screened against CHIKV infection of HuH-7 cells using the resazurin reduction assay, and the cell toxicity was also measured in non-infected cells. Seventy-two compounds showing ≥50% inhibition property against CHIKV at 10 µM were selected as primary hits. Four compounds having a benzofuran core scaffold (CND0335, CND0364, CND0366 and CND0415), one pyrrolopyridine (CND0545) and one thiazol-carboxamide (CND3514) inhibited CHIKV-associated cell death in a dose-dependent manner, with EC50 values between 2.2 µM and 7.1 µM. Based on image analysis, these 6 hit compounds did not inhibit CHIKV replication in the host cell. However, CHIKV-infected cells manifested less prominent apoptotic blebs typical of CHIKV cytopathic effect compared with the control infection. Moreover, treatment with these compounds reduced viral titers in the medium of CHIKV-infected cells by up to 100-fold. In conclusion, this cell-based high-throughput screening assay using resazurin, combined with the image-based high content assay approach identified compounds against CHIKV having a novel antiviral activity - inhibition of virus-induced CPE - likely by targeting kinases involved in apoptosis.

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Section: Discussionmentioning

confidence: 99%

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

et al. 2013

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“…[19][20][21][22][23][24] MTT and other tetrazolium-based assays are based on the cleavage and reduction of the tetrazolium ring to blue formazan crystals by the mitochondrial dehydrogenases. 22,23 These crystals can be solubilized and quantified in the cell lysate as a measure for metabolic cell activity.…”

Section: Introductionmentioning

confidence: 99%

Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

Sonnaert

Papantoniou

Luyten

et al. 2015

Tissue Engineering Part C: Methods

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As the fields of tissue engineering and regenerative medicine mature toward clinical applications, the need for online monitoring both for quantitative and qualitative use becomes essential. Resazurin-based metabolic assays are frequently applied for determining cytotoxicity and have shown great potential for monitoring 3D bioreactor-facilitated cell culture. However, no quantitative correlation between the metabolic conversion rate of resazurin and cell number has been defined yet. In this work, we determined conversion rates of Presto BlueÔ, a resazurin-based metabolic assay, for human periosteal cells during 2D and 3D static and 3D perfusion cultures. Our results showed that for the evaluated culture systems there is a quantitative correlation between the Presto Blue conversion rate and the cell number during the expansion phase with no influence of the perfusion-related parameters, that is, flow rate and shear stress. The correlation between the cell number and Presto Blue conversion subsequently enabled the definition of operating windows for optimal signal readouts. In conclusion, our data showed that the conversion of the resazurin-based Presto Blue metabolic assay can be used as a quantitative readout for online monitoring of cell proliferation in a 3D perfusion bioreactor system, although a system-specific validation is required.

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“…Moreover, inconsistencies between measurements with various assays are not uncommon [23][24][25][26][27], and they arise from individual characteristics of the measuring techniques. It has been reported that the MTT assay, which is the most commonly used method, has several disadvantages, including poor linearity with the cell number, sensitivity to environmental conditions, and dependence of the cell metabolism on formazan, which is formed during enzymatic conversion of a dye in the cell mitochondria, as well as in other cell organelles [28].…”

Section: Resultsmentioning

confidence: 99%

Endothelialization of Polyethylene Terephthalate Treated in SO2 Plasma Determined by the Degree of Material Cytotoxicity

Vesel¹,

Recek²,

Motaln³

2017

Plasma

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Improving the biocompatibility of polyethylene terephthalate (PET) vascular grafts is an important task for avoiding thrombus formation. Therefore, SO 2 plasma at various treatment periods were used to modify PET surface properties by forming sulfate functional groups. These groups were shown to act antithrombogenically, ensuring good hemocompatibility of the materials, although the biocompatibility of such materials still remains a mystery. For this reason, the adhesion and viability of HUVEC cells on SO 2 plasma-modified PET surfaces were studied, and the possible toxicity of the tested material was determined using two different assays, MTT (metabolic activity assay) and SRB (in-vitro toxicology assay). Changes in chemical composition, morphology and wettability were determined as well. Improved endothelialization was observed for all plasma-treated samples, with the most optimal being the sample treated for 80 s, which can be explained by it having the best combination of surface functionalization, roughness and morphology. Furthermore, toxicity was observed to some extent on the sample treated for 160 s, indicating the lowest cell density among the plasma-treated samples. X-ray photoelectron spectroscopy showed increased oxygen and sulfur content on the surface, which was independent on treatment time. Surface roughness of the plasma-treated samples increased, reaching its maximum after 80 s of treatment, and decreased thereafter.

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Comparison of XTT and Alamar blue assays in the assessment of the viability of various human cancer cell lines by AT-101 (−/− gossypol)

Cited by 38 publications

References 14 publications

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

Endothelialization of Polyethylene Terephthalate Treated in SO2 Plasma Determined by the Degree of Material Cytotoxicity

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