The harvested cells were seeded into the 96-well plate at the density of 3 × 103 cells per well and maintained at 37°C with 5% CO2 overnight. When the confluence reached 80–90%, the cells were treated for 24 h with different concentration of quercetin (Que, 0, 12.5, 25, 50, and 100 μM) [20 (link)], followed by addition 200 μM H2O2 treatment. The cells were then treated with addition of 10 μl of cell counting kit-8 (CCK-8) solution (Solarbio) and cultured for 4 h. Then, the cells were subjected to a microplate reader (Allsheng) to detect the optical density of cell at 450 nm.
Cell counting kit 8 cck 8 solution
Manufactured by Solarbio
Sourced in China
The Cell Counting Kit-8 (CCK-8) solution is a colorimetric assay for the determination of cell viability and proliferation. It utilizes the highly water-soluble tetrazolium salt WST-8, which is reduced by cellular dehydrogenases to give a yellow-colored formazan dye. The amount of the formazan dye generated is directly proportional to the number of living cells.
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9 protocols using cell counting kit 8 cck 8 solution
1
Quercetin Cytotoxicity Assay in H2O2-Treated Cells
2
Evaluating Doxorubicin Cytotoxicity in Cells
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Cells (3 × 103) were seeded into 96-well plates and stimulated with escalating doses of DXR for 48 h. At the same time, transfected DXR-resistant cells (3 × 103) were cultured in 96-well plates for 0 h, 24 h, 48 h or 72 h. Subsequently, cells were interacted with 10 μL Cell Counting Kit-8 (CCK-8) solution (Solarbio) for 2 h. Then, the optical density was measured at 450 nm using a Multi-Mode Reader (BioTek, Burlington, VT, USA). The half maximal inhibitory concentration (IC50) of DXR was the concentration of DXR when cell viability was reduced to 50%.
3
Constructing Tissue-Engineered Bone with BMSCs/XACB
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Based on our previous research3 (link), we cut XACB into small cylinders 5 mm in diameter at the bottom and 5 mm high and immersed them in complete L-DMEM for 24 h. We added BMSCs suspension (1 × 107/mL) to XACB, allowed the cells to adhere for 3 h in an incubator (37 °C, 5% CO2), slowly added complete L-DMEM along the wall of the culture plate, and continued to culture the mixture in the incubator (37 °C, 5% CO2) to construct tissue-engineered bone (BMSCs/XACB). Each day for the next week, we took five pieces of BMSCs/XACB out of the incubator, collected cells from them via digestion and centrifugation, and added 10 μL of Cell Counting Kit-8 (CCK-8) solution (Solarbio) to each well. We measured the absorbance value (450 nm) until day 7, when we drew the growth curve based on culture time and absorbance value. We cultured XACB and BMSCs on day 6 and observed the growth of BMSCs on the surface of XACB using a SEM (Hitachi).
4
Cell Viability Assay with CCK-8
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Cardiomyocytes were plated in a 96-well plate at a density of 2×104 cells/well. Next, 10 µl Cell Counting Kit-8 (CCK-8) solution (Beijing Solarbio Science & Technology Co., Ltd.) was added and cells were cultured for 4 h at 37°C in the dark. Finally, the absorbance was measured at 450 nm using a Universal Microplate Spectrophotometer (Bio-Rad Laboratories, Inc.).
5
Cell Viability Quantification via CCK-8
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The cardiomyocytes were seeded in a 96-well plate at a density of 2 × 104 cells/well. Next, the cells were added to 10 μl Cell Counting Kit-8 (CCK-8) solution (Solarbio, Beijing, China) and incubated for 4 h at 37°C in the dark. Finally, the absorbance was measured at 450 nm using a Universal Microplate Spectrophotometer (Bio-Rad, CA, USA).
6
Neuronal Cell Viability Assay
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Neurons cultured in 96-well plates at a concentration of 1 × 105 cells/well were treated with ICT (0.1, 0.3, and 1 μM) and glutamate as previously described. Fourteen hours after the treatment, 10 μL of Cell Counting Kit-8 (CCK-8) solution (Solarbio) was added to each well according to the manufacturer’s instructions. After incubation at 37°C with 5% CO2 for 3 h, the absorbance value of each well was measured at a 490 nm wavelength by a Multi-Mode Microplate Reader (Biotek Instruments, INC Winooski, Vermont, United States). The absorbance value of each well was calibrated by subtracting the value of an empty well.
7
Evaluating H9C2 Cell Viability
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H9C2 cells were seeded at a density of 5,000 cells per well in 96-well culture plates lined with different hydrogels. When the cells reached 70% to 80% density, 0.1 mM H2O2 was added and treated for 24 h. Subsequently, 10 μl of Cell Counting Kit-8 (CCK-8) solution (Solarbio, China) was added to each well and treated for 2 h. The absorbance was measured at a wavelength of 450 nm.
8
Cell Viability Assessment using CCK-8
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The treated or untreated SK-N-SH cells (3×103) were plated into 96-well plates. Subsequently, cells were incubated for 3 h after adding 10 μL Cell Counting Kit-8 (CCK-8) solution (Solarbio, Beijing, China). Next, Microplate Reader (BioTek, Burlington, VT, USA) was used to evaluate cell viability by measuring absorbance at 450 nm.
9
Cell Viability Assay in Cardiomyocytes
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The cardiomyocytes were seeded in a 96-well plate at a density of 2 × 10 4 cells/well. Next, the cells were added to 10 µl Cell Counting Kit-8 (CCK-8) solution (Solarbio, Beijing, China) and incubated for 4 h at 37 °C in the dark. Finally, the absorbance was measured at 450 nm using a Universal Microplate Spectrophotometer (Bio-Rad, CA, USA).
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