The cells were seeded in 96-well plates. After overnight culture, the cells were treated with 0, 25, 50, 100, 200, and 400 μg/mL of PM2.5 or 5, 10, 20, 30, 40, and 50 mM of glucose for 24 h. The cytotoxicity of PM2.5 or glucose in MH-S cells was determined by two different methods: CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay (MTS assay) (Promega, Madison, WI, USA) and alamarBlue™ assay (Invitrogen, Eugene, OR, USA) according to the manufacturer’s instructions. To observe whether there was cytotoxicity with PM2.5 treatment in a high glucose environment, the cells were pretreated with 30 mM glucose for 18 days followed by 25 or 50 μg/mL of PM2.5 treatment for another 24 h. Mannitol was used as an osmolality control.
Celltiter 96 aqueous non radioactive cell proliferation assay mts assay
Manufactured by Promega
Sourced in United States
The CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay (MTS assay) is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The assay measures the bioreduction of a tetrazolium compound (MTS) into a colored formazan product by viable cells. The amount of formazan produced is directly proportional to the number of living cells in culture.
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Lab products found in correlation
Alamarblue assay, by Thermo Fisher Scientific (2 mentions)
Optical power meter, by Thorlabs (2 mentions)
Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Ldh cytotoxicity detection kit, by Promega (1 mentions)
Infinite f200, by Tecan (1 mentions)
Hacat cells, by American Type Culture Collection (1 mentions)
Hdf cells, by American Type Culture Collection (1 mentions)
Mouse 3t3 fibroblasts, by American Type Culture Collection (1 mentions)
16 protocols using celltiter 96 aqueous non radioactive cell proliferation assay mts assay
1
Cytotoxicity of PM2.5 and Glucose in Macrophages
2
Cytotoxicity of Nano-CuO and Nano-TiO2
3
Cell Cytotoxicity Assay Protocol
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Cell cytotoxicity was assessed through CellTiter 96® Aqueous Non-Radioactive Cell Proliferation Assay (MTS Assay) (Promega, USA) as per manufacturer's instructions [33 ]. Cell numbers were adjusted after cell counts (MCF-7: 8.0x103 cells/well; HT-29: 1.0x104 cells/well; HCT-116: 2.0x104 cells/well; MDA-MB-231 and Reh: 2.5x104 cells/well) using their respective complete medium without phenol red. 90μL of cell solutions were inoculated into a 96-well plate and incubated overnight. After incubation, 10μL of different concentrations of various extracts and fractions were introduced into respective well. 5-fluorouracil (5-FU) (donated by Penang General Hospital, Malaysia) was used as reference drug for comparison. After 48 hours, 20μL of MTS reagents was added into each well followed by incubation for another 2-5 hours (HT-29: 2 hours; HCT-116: 3 hours; MCF-7, MDA-MB-231, and Reh: 5 hours). The absorbance value for each well was measured at 490nm and 630nm (reference wavelength) using a microplate reader. The percentage of inhibition for each extract and fraction was calculated as follows: whereby
An = absorbance of negative control
As = absorbance of test samples (extracts, fractions, or 5-FU)
An = absorbance of negative control
As = absorbance of test samples (extracts, fractions, or 5-FU)
4
Regorafenib Cytotoxicity in Cell Lines
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Cells were seeded at 5,000 or 25,000 per well in 96-well plates. After 24 hours regorafenib was added to final concentrations between 0.01–100 μmol/L for 72 hours. Cell viability was determined using the CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS assay; Promega) according to manufacturer’s instructions. Each concentration and time point was tested in quadruplet and all experiments in triplicates. Means and standard deviations were calculated from quadruple optical density (OD) measurements. Viability of treated cells was compared to that of untreated cells and drug concentrations which inhibited cell growth by 50% were calculated as described previously [42 (link)].
5
Optimizing Biomimetic Bone Scaffolds for hMSC Growth
6
Cell Viability Assay with MM80
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3T3 mouse fibroblasts and HEK293 cells were cultured as previously described [13 (link)]. The cells were seeded in a 96-well plate at a density of 5,000 cells per well. The plates were incubated at 37 °C under a humidified atmosphere of 5% CO2 for 24 h. Meanwhile, lyophilised MM80 was ground in DMEM using mortar and pestle to obtain a suspension with the desired concentration. The cell culture medium was replaced with 100 μL of the suspension. After 5 h incubation at 37 °C, the suspension in each well was replaced with the fresh cell culture medium. The CellTiter 96 Aqueous Non-radioactive Cell Proliferation Assay (MTS assay; Promega Corp., Madison, WI, USA) was performed according to the manufacturer’s instructions, either immediately or after 24 h of post-treatment incubation to determine the cell viability (%) in each well.
7
Cytotoxicity Evaluation of Biomaterials
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HEK293 cells and 3T3 mouse fibroblasts were cultured as previously described.23 24 hours before the experiment, the cells were seeded in 96-well plates at a density of 5000 cells per well. The plates were put under a humidified atmosphere of 5% CO2 at 37 °C. An appropriate amount of CS, GE, or lyophilized gels (including G100, G050 and G000) was ground in the fresh cell culture medium using mortar and pestle to obtain a suspension with a desired concentration. The suspension was then filtered by using a microfilter with the pore size of 0.45 μm. During the assay, the medium in each well was replaced with 100 μL of the filtrate. After 5 hour incubation at 37 °C under a humidified atmosphere of 5% CO2, the medium in each well was replaced with the fresh cell culture medium. The viability of cells in each well was examined by using the CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay (MTS assay; Promega Corp., Madison, WI), according to the manufacturer's instructions, either immediately or after 24 hours of post-treatment incubation.
8
Cellular Viability and Proliferation Assays
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Changes in cellular viability were investigated 4 days post-differentiation, using the CyQUANT lactate dehydrogenase (LDH) Cytotoxicity Assay (LDH assay) (Thermo Fisher, C20300). Furthermore, changes in cellular proliferation were investigated 4 days post-differentiation, using the CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay (MTS assay) (Promega, G5421). Both assays were conducted according to the manufacturer’s instructions.
9
Cytotoxicity and Metabolic Activity of HRV-Infected A549 Cells Treated with LL-37
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The LDH assay was performed using the LDH cytotoxicity detection kit (Promega) according to the manufacturer’s instructions. Metabolic activity was assessed by use of the CellTiter96 Aqueous Non-Radioactive Cell Proliferation Assay (MTS assay) (Promega) according to the manufacturer’s instructions. Briefly, A549 cells were seeded at 2 × 104 per well in a 96-well plate and cultured overnight at 37 °C. Cells were infected with HRV at an MOI of 10 (with or without LL-37). Cells were also incubated with peptide alone. After incubation of cells with HRV and/or peptide at 33 °C, supernatant was aspirated and LDH activity was measured. Cells were resuspended in DMEM containing MTS reagent and metabolic conversion of MTS reagent was assessed after 2 h by measuring the optical density at 595 nm. Controls included untreated cells as negative control, and cells treated with Triton-X as positive control for reduction in metabolic activity.
10
Doxorubicin Cytotoxicity in CSF-1R Populations
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Cells were plated in triplicate at 2,500 (DD-1), 5,000 (COSB) or 10,000 (AS5) cells per well in 100 μL of culture medium and exposed to increasing concentrations of doxorubicin (Bedford Laboratories, Bedford, OH). Cell viability was determined 72 hours later using the colorimetric Cell Titer 96® Aqueous Non-Radioactive Cell Proliferation Assay (MTS Assay; Promega, Madison, WI) according to the manufacturer’s instructions. In order to compare the percent viability between the CSF-1Rlow and the CSF-1Rhigh populations and reduce artifacts due to potential differences in metabolic properties between cell populations, standard curves were generated for each CSF-1R population in each cell line, and the cell viability was determined based upon the total cell number from the standard curve. A separate standard curve for the CSF-1Rlow and the CSF-1Rhigh populations was generated for each experiment.
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