Cells were cultured in 96-well plates for 24 hours, before serum starvation for 16 hours. Test groups were cultured in ADSC-CM for another 48 hours in triplicate. Control groups were cultured in normal medium. Cell viability was then measured by MTS assay (CellTiter96 AQueous Assay; Promega, France).
Celltiter 96 aqueous assay
Manufactured by Promega
Sourced in United States, France, Germany
The CellTiter 96® AQueous Assay is a colorimetric method for determining the number of viable cells in cell proliferation or cytotoxicity assays. It utilizes a novel tetrazolium compound that produces a colored formazan product when reduced by viable cells.
Automatically generated - may contain errors
Lab products found in correlation
Mts tetrazolium reagent, by Promega (4 mentions)
El808, by Agilent Technologies (4 mentions)
Infinite m1000 pro microplate reader, by Tecan (3 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Automatic handheld cell counter, by Merck Group (2 mentions)
Spektramax, by BMG Labtech (2 mentions)
Spectramax m2, by Molecular Devices (2 mentions)
96 well microtiter plates, by Brandel Inc (2 mentions)
Mc3t3 e1, by American Type Culture Collection (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Fetal calf serum (fcs), by Merck Group (1 mentions)
Dmem f12, by Corning (1 mentions)
Multimode microplate reader, by Tecan (1 mentions)
Victor3 multilabel counter, by PerkinElmer (1 mentions)
Sh sy5y cell line, by Thermo Fisher Scientific (1 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions)
Prism software, by GraphPad (1 mentions)
Eμ myc mice, by Jackson ImmunoResearch (1 mentions)
Trametinib, by Cayman Chemical (1 mentions)
51 protocols using celltiter 96 aqueous assay
1
Cell Viability Assay with ADSC-CM
2
Splenocyte Proliferation Assay
3
Evaluating Osteoblastic Cell Viability with MSNs
4
Curcumin and C-150 Cytotoxicity in Glioblastoma
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Glioblastoma cells (U373 MG, U87 MG, GBM1-GBM6 cell lines were a kind gift from Balazs Hegedus, Semmelweis University, Hungary, [53 (link)] while the U251 MG cell line was a gift from Szabolcs Bellyei, University of Pecs, originally obtained from American Type Culture Collection, Manassas, VA) were grown at 37°C under 5% of CO2 and 100% humidity in DMEM and RPMI medium supplemented with 10% fetal calf serum (FCS) (Sigma-Aldrich), and penicillin-streptomycin antibiotics. For cytotoxicity assays, 10,000 cells were seeded into each well of 96-well cell culture plates in culture medium containing 10% FCS. Effects of curcumin and C-150 were recorded 48 h after treatment. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazo-lium) reagent was applied to drug treated and control (0.2% DMSO) cells (CellTiter 96® AQueous Assay, Promega, Madison, WI) according to the manufacturer’s protocol.
5
Optimizing PDCL Cell Density and Cytotoxicity
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The optimum cell density of each PDCL was established using the MTS, CellTiter 96® Aqueous Assay (Promega®) and viability was measured 8 days post-treatment. PDCLs were treated with increasing concentrations of veliparib (1–10 μM); TMZ (100–300 μM) and/or radiation (1–4 Gy) to determine the cytotoxic effects of the chemotherapeutic drugs, and the half-maximal inhibitory concentration (IC50).
6
Determining 5-FU Sensitivity in CRC-stem Cells
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Tu11, Tu14, Tu27, Tu28, and Tu42 CRC-stem like cells were cultured in Dulbecco’s Modified Eagle Medium:Nutrient Mixture F-12 (DMEM/F12, Corning, cat. no. 10090CV; Fisher Scientific, Pittsburgh, PA, USA) containing 1% insulin-transferrin-selenium (ITS, Corning, cat. no. 25800CR) and 0.5% fetal bovine serum (FBS, Atlanta Biologicals, cat. no. S1245OH; Fisher Scientific). To establish 5-FU IC50, cells were seeded onto 96-well plates (10,000 cells/well) in 100 µL of culture medium overnight, and then treated in triplicate with serial dilutions of 5-FU (10–250 µM) (Sigma-Aldrich, cat. no. F6627-1G; Saint Louis, MO, USA). The number of viable cells was measured using the MTS-based CellTiter 96 AQueous Assay (Promega, cat. no. G3582; Madison, WI, USA) 72 h after drug exposure. 20 µL of the MTS reagent were added to each well, and the absorbance was recorded at 490 nm after 4 h incubation at 37 °C. Data were expressed as mean percentage (±SD) of cell numbers relative to control culture.
7
Cell Viability Assay Using CellTiter 96 AQ
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The CellTiter 96 AQueous assay (Promega, USA) was used to determine cell viability (8 (link)). The CellTiter 96 AQueous Assay is composed of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and phenazine methosulfate (PMS). RAW264.7 or HaCaT were seeded to 96-well plates (1 × 104 cells/well) for 18 to 24 h. Fresh media with or without different concentrations of the test peptide were added slightly to the wells and incubated for another 24 h. The vehicle group was treated with fresh medium containing 5% water. After incubation with tested peptide, 20 μL of MTS-PMS reagent was added to each well and incubated for 1 to 4 h. The absorbance of each well was measured at 492 nm in a multimode microplate reader (Tecan, Switzerland). The experiments were repeated three times independently.
8
Cytotoxicity of Allium Extracts
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Cytotoxicity of Allium extracts was determined on three cancer cell lines: cervical cancer cell line (HeLa), human colon cancer cell line (HCT116), and human osteosarcoma cell line (U2OS). The cells were a kind gift from Janoš Terzić laboratory at the School of Medicine, University of Split. The cytotoxicity of the extracts was determined by using the MTS-based CellTiter 96® Aqueous assay (Promega). Cells were grown in an incubator at 37 °C and 5% CO2, until they reached 80% confluency. Cells were counted with a handheld automated cell counter (Scepter, Merck), and 5000 cells/well were seeded in 96-well plates containing a serial dilution of both extracts. Cells were further grown for an additional 48 h, after which 20 µL of MTS tetrazolium reagent (Promega) was added to each well. DMSO was used as vehicle control. After 3 h of incubation, the absorbance at 490 nm was measured by a 96-well plate reader (Bio-Tek, EL808). The measurements were performed in quadruplets, and IC50 values were calculated from three independent experiments, using GraFit 6 data analysis software (Erithacus, East Grinstead, UK).
9
Doxorubicin and LHRH Analog Cytotoxicity
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Cells were seeded in 96-well plates at 2,500 cells/well density in complete growth medium and were incubated for 24 hrs. After 24 hrs, culture medium was replaced by serum-free medium for another 24 hrs. This was followed by the addition of 100 nM or 250 nM of unconjugated doxorubicin, [D-Lys(6)]LHRH), their combination, or AEZS-108 in medium containing 0.5% heat-inactivated serum; the cells were then incubated for a further 48 h. At the end of the treatment the relative number of viable cells was measured using MTS assay (CellTiter 96 AQueous Assay; Promega, Madison, WI) following manufacturer's instructions. Absorbance was measured at 490 nm in a Victor 3 Multilabel Counter (Perkin-Elmer, Waltham, MD). Experiments were performed in hexaplicate. Values were expressed relative to the control.
10
Antiproliferative Effects of P. ramosissima Extract on Cancer Cell Lines
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The antiproliferative analysis of methanol extract of P. ramosissima on three cancer cell lines, cervical cancer cell line (HeLa), human colon cancer cell line (HCT116), and human osteosarcoma cell line (U2OS) was performed according to the protocol described in our previously published papers [39 (link)] using the MTS-based CellTiter 96® Aqueous Assay (Promega, Madison, WI, USA). Cells were donated by prof. Janoš Terzić from the School of Medicine, University of Split. Cells were grown in a CO2 incubator at 37 °C and 5% CO2 until they reached 80% confluency. They were further counted using the automatic handheld cell counter (Merck, Darmstadt, Germany), 5000 cells per well were seeded in 96-well plates and treated with serially diluted methanolic extract of P. ramosissima. The cells were cultured for an additional 48 h, after which 20 µL of MTS tetrazolium reagent (Promega, Madison, WI, USA) was added to each well and left for 3 h in the incubator at 37 °C and 5% CO2. The absorbance was measured at 490 nm using a 96-well plate reader (Bio-Tek, EL808, Winooski, VT, USA). Measurements were performed in four replicates for each concentration and IC50 values were calculated from three independent experiments.
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