Cell viability was assessed using a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) reagent (CellTiter96® Aqueous One Solution Cell Proliferation Assay, Promega Madison, WI, USA). Primary human trabecular meshwork (hTM) cells (30,000 cells/well/200 µL) were seeded in 96-well plates to confluency. The cells were serum-deprived for 24 h and were exposed to different concentrations of SA-2 (1 µM, 10 µM, 100 µM, and 1000 µM). After 24 h of incubation at 37 °C, cell viability was measured using an MTT assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega Madison, WI, USA) following the manufacturer’s protocol. Experiments were repeated twice with three technical replicates, and the percentage of viable cells was determined by normalizing to the untreated control group.
Celltiter 96 aqueous one solution cell proliferation assay
Manufactured by Promega
Sourced in United States, Italy, Germany, United Kingdom, Spain, Switzerland, Sweden, China, Japan, France, Australia, Finland
The CellTiter 96® AQueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The assay reagent contains a tetrazolium compound that is bioreduced by cells into a colored formazan product that is soluble in tissue culture medium, allowing the determination of viable cells.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (39 mentions)
Synergy ht, by Agilent Technologies (29 mentions)
Microplate reader, by Bio-Rad (22 mentions)
Infinite m200, by Tecan (17 mentions)
Infinite m200 pro, by Tecan (16 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (15 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (14 mentions)
Celltiter glo luminescent cell viability assay, by Promega (14 mentions)
96 well plate, by Corning (14 mentions)
Prism 8, by GraphPad (14 mentions)
Cisplatin, by Merck Group (12 mentions)
Microtest 96, by BD (12 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (12 mentions)
Epoch microplate spectrophotometer, by Agilent Technologies (11 mentions)
Elx800, by Agilent Technologies (11 mentions)
Imark microplate reader, by Bio-Rad (11 mentions)
Spectramax m2, by Molecular Devices (11 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (11 mentions)
Streptomycin, by Thermo Fisher Scientific (11 mentions)
Imark microplate absorbance reader, by Bio-Rad (10 mentions)
2 189 protocols using celltiter 96 aqueous one solution cell proliferation assay
1
Evaluating Trabecular Meshwork Cell Viability
2
Cell Viability Assay Protocols
3
Cytotoxicity Evaluation of Compounds
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The cytotoxicity of the
compounds was determined using the MTS cell proliferation assay (CellTiter
96 AQueous One Solution Cell Proliferation Assay, Promega, Wisconsin,
USA), according to the manufacturer's protocol on the HEK293
cell
line, cotransfected with the hTLR7 gene (InVivoGen). Cells were seeded
in 96-well plates (1.5 × 104 cells/well). After 24
h incubation at 5% CO2 and 37 °C, the cells were treated
either with DMSO (negative control) or with solutions of the tested
compounds in DMSO. After 24 h, MTS reagent (CellTiter 96 AQueous One
Solution Cell Proliferation Assay, Promega, Wisconsin, USA) was added.
Cells were incubated for another 2 h, and then absorbance was measured
at 490 nm.
compounds was determined using the MTS cell proliferation assay (CellTiter
96 AQueous One Solution Cell Proliferation Assay, Promega, Wisconsin,
USA), according to the manufacturer's protocol on the HEK293
cell
line, cotransfected with the hTLR7 gene (InVivoGen). Cells were seeded
in 96-well plates (1.5 × 104 cells/well). After 24
h incubation at 5% CO2 and 37 °C, the cells were treated
either with DMSO (negative control) or with solutions of the tested
compounds in DMSO. After 24 h, MTS reagent (CellTiter 96 AQueous One
Solution Cell Proliferation Assay, Promega, Wisconsin, USA) was added.
Cells were incubated for another 2 h, and then absorbance was measured
at 490 nm.
4
Cytotoxicity of Gold Nanoparticles on Colon Cancer
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Cytotoxic activity of AuS NPs against human colon cancer cells (SW480 and SW620) was determined using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI, USA). Briefly, cells were cultured in flat-bottom 96-well plates (Sarstedt, Numbrecht, Germany) at a density of 1 × 104/well in DMEM medium containing 10% FBS. After 24 h, 20 μL of 3.88 × 10−3 mg/mL Au NPs solution was added to the cells. After additional 24 h of culture, 20 µL of MTS (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega) dye solution was added per well and incubated for 2 h. The quantity of formazan product, directly proportional to the number of living cells in culture, was detected by absorbance measurements at 490 nm with a 96-well plate reader (Spark® Tecan, Mannedorf, Switzerland).
5
Deguelin Cytotoxicity Assay in NuFF-1 Cells
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Cytotoxic effects of deguelin were measured by treating confluent NuFF-1 cells with varying concentrations of deguelin for 3 days (d), as indicated. Cell proliferation was measured using the CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega, Madison, WI, USA), in accordance to the manufacturer’s instructions. Absorbance was read at 490 nm using a Cytation 3 cell imaging plate reader (BioTek, Winooski, VT, USA). Data is plotted as percent inhibition relative to vehicle (DMSO)-treated cells. Cell doubling was assessed by seeding NuFF-1 cells at 25% confluence and treating with varying concentrations of deguelin, as described in the text. After two cell doublings, cell proliferation was measured using CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega), as described above. Drug-treated cells were normalized to vehicle (DMSO)-treated cells.
6
Evaluating Nanomaterial Cytotoxicity on Colon Cancer
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Cytotoxic activity of NPs, cPt, laser irradiation and combination of these three factors against human colon cancer cells (SW480 and SW620) was determined using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI). Briefly, the cells were cultured in flat-bottom 96-well plates (Sarstedt, Numbrecht, Germany) at a density 1 × 104 per well in DMEM medium containing 10% FBS. After 24 h, 20 μL of 50 μg mL−1 functionalized Fe3O4@SiO2@Au NPs as well as non-functionalized solutions were added to the cells. After additional 24 hours of culture, 20 μL of MTS (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega) dye solution was added per well and incubated for 1.5 h. The quantity of formazan product, directly proportional to the number of living cells in culture, was detected by absorbance measurements at 490 nm with a 96-well plate reader (Spark® Tecan, Mannedorf, Switzerland).
7
Cytotoxic Effects of PtAu Nanoraspberries on Colon Cancer Cells
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The cytotoxic activity of PtAu nanoraspberries against human colon cancer cells (SW480 and SW620) was determined by using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI). Briefly, the cells were cultured in flat-bottom 96-well plates (Sarstedt, Numbrecht, Germany) at a density of 1 × 104/well in DMEM medium containing 10% FBS. After 24 h, 20 μL of PtAu NRs solution was added to the cells. After additional 24 h of culture, 20 µL of MTS (CellTiter 96® Aqueous One Solution Cell Proliferation Assay, Promega) dye solution was added per well and incubated for 2 h. The quantity of formazan product, directly proportional to the number of living cells in culture, was detected by absorbance measurement at 490 nm with a 96-well plate reader (Spark® Tecan, Mannedorf, Switzerland). SW480 and SW620 cancer cell lines cultured without PtAu NRs and without irradiation were used as control samples.
In the present study the following samples were investigated (Table1 ).
In the present study the following samples were investigated (Table
Investigated samples
| Control | SW480 | SW620 |
|---|---|---|
| Investigated samples | SW480 + PtAu NRs | SW620 + PtAu NRs |
| SW480 + laser 650 nm and 808 nm | SW620 + laser 650 nm and 808 nm | |
| SW480 + PtAu NRs + laser 650 nm and 808 nm | SW620 + PtAu NRs + laser 650 nm and 808 nm |
8
MDA-MB231 Cell Proliferation Under DMEM and SCM
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For MDA-MB231 cell proliferation under DMEM conditions, cells were seeded in DMEM plus 10% FBS and 1% Penicillin/Streptomycin at a density of 1×105 cells per well in 6 well plate, and stained with trypan blue and counted for viable cells everyday for 5 days. Alternatively, cells were seeded at a density of 3,000 cells per well in 96 well tissue culture plate (08-772-3; Fisher), and cell growth was determined everyday for 4–5 days by CellTiter96 AQueous One Solution Cell Proliferation Assay as instructed by the manufacturer (Promega). For cell growth under SCM conditions, cells were seeded in SCM at a density of 3,000 cells per well in 96 well non-treated microplate (12-565-226; Fisher), and supplemented with fresh human EGF (25 ng/ml), basic FGF (25 ng/ml) and Insulin (5 ug/ml) every 2 days. Cell growth was determined at indicated time points by CellTiter96 AQueous One Solution Cell Proliferation Assay as instructed by the manufacturer (Promega). Triplicates were performed for each time point of the growth curve.
9
Quantifying Cell Proliferation Inhibition
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The MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] Assay (CellTiter 96 AQueous One Solution Cell Proliferation Assay; Promega, Madison, WI) was performed as described in the manufacturer's instructions. Briefly, cells (HT-29 or T4056) were initially passaged, and counted in a hemocytometer for the purpose of calculating the appropriate seeding. Cells were plated at the concentration of 1.0 × 10 3 cells per well and the inhibition of cell proliferation was assayed using the MTS assay (CellTiter 96 AQueous One Solution Cell Proliferation Assay, Promega). Briefly, 20 μL of soluble extract sample were added to the cells (1.0 × 10 3 in 80 μL of complete medium). After 72 h of exposure of the sample to the cells, the MTS reagent (20 μL/100 μL of medium) was added and incubated for an additional 4 h under the same conditions. The absorbance of formazan was measured at 495 nm and unstimulated cells were used as a control. All assays were performed in triplicates and the results expressed as mean values ± standard error. The percentage of cell inhibition of colon cancer and normal colon cells was calculated (Kim et al., 2000) from a ratio of treatment values compared with controls as follows:
Proliferative inhibition (%) = (Treatment A 495 /Control A 495 ) × 100, where A 495 = absorbance at 495 nm.
Proliferative inhibition (%) = (Treatment A 495 /Control A 495 ) × 100, where A 495 = absorbance at 495 nm.
10
Cytotoxicity of Gold Nanoparticles on A549 and Du145 Cells
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A549 and Du145 were seeded in 96-well plates with 3,000 cells per well and incubated at 37°C with 5% CO2 in a humidified environment. After 24 hours of incubation, the cells were treated with various concentration of AuNPs ranging from 0.0 (control) to 4.0 mM.
MTS (3 - (4, 5 - dimethylthiazol - 2 - y l) - 5 - (3 -carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay was performed using CellTiter 96® AQueous One Solution Cell Proliferation Assay (Promega Corporation, Fitchburg, WI, USA); 24 and/or 48 hours after treating with AuNPs, the medium was removed and 100 µL of cultured medium supplied with 10 µL of CellTiter 96® AQueous One Solution Cell Proliferation Assay was added to the cells. Immediately after adding MTS, the optical absorbance of the formazan was measured at 490 nm using a CLARIOstar® microplate reader (BMG LABTECH, Mornington, Australia) to determine the background (BG). The plates were then incubated for 1 hour followed by measuring the optical absorbance. The results are expressed as a percentage relative to the control groups (Equation 2 ).
The cell viability was measured at 24 and 48 hours after inclusion of AuNPs. Cytotoxicity assay indicated probabilistic toxic effects of AuNPs on cells without any exposure to radiations.
MTS (3 - (4, 5 - dimethylthiazol - 2 - y l) - 5 - (3 -carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay was performed using CellTiter 96® AQueous One Solution Cell Proliferation Assay (Promega Corporation, Fitchburg, WI, USA); 24 and/or 48 hours after treating with AuNPs, the medium was removed and 100 µL of cultured medium supplied with 10 µL of CellTiter 96® AQueous One Solution Cell Proliferation Assay was added to the cells. Immediately after adding MTS, the optical absorbance of the formazan was measured at 490 nm using a CLARIOstar® microplate reader (BMG LABTECH, Mornington, Australia) to determine the background (BG). The plates were then incubated for 1 hour followed by measuring the optical absorbance. The results are expressed as a percentage relative to the control groups (
The cell viability was measured at 24 and 48 hours after inclusion of AuNPs. Cytotoxicity assay indicated probabilistic toxic effects of AuNPs on cells without any exposure to radiations.
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