The viability of HDFs was evaluated using MTT assay. HDFs were seeded in 96-well plates at 5 × 104 cells/well in 200 μL of complete media and incubated for 24 hours in the incubator to allow cell adhesion. The cells were exposed to ultrasound beam according to the procedures mentioned above and then returned to the humidified incubator for other 24 hours. Then, supernatant medium was discarded and 100 μL fresh serum-free medium was added. Then, we added a 50 μL MTT solution (0.5 mg/mL) (Bio idea, Tehran, Iran) to each well and incubated plates for an additional 4 hours in dark conditions. After that, we removed the medium and added 200 μL DMSO (Bio idea, Tehran, Iran) to each well in order to dissolve the purple formazan precipitate. Absorbance was measured using an ELISA microplate reader (Anthos 2020, Biochrom Ltd, and UK) at a wavelength of 570 nm. We calculated the percentage of viability according to the following formula: Viability (%) = (average OD (optical density) in the treated sample/ average OD in the control sample) × 100.
Mtt solution
Manufactured by Bioidea
MTT solution is a reagent used in cell-based assays to measure cell viability and proliferation. It is a colorimetric assay that relies on the conversion of a tetrazolium salt (MTT) into a formazan product by the mitochondrial enzymes of living cells. The resulting color change can be quantified using a spectrophotometer, providing an indication of the metabolic activity and, consequently, the number of viable cells in the sample.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mtt solution
1
Evaluating HDF Viability Using MTT Assay
2
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After seeding 5 × 103 cells in each 96-well plate, they were incubated in 5% CO2 at 37 °C. 24 h after transfection, the MTT assay was carried out. Following the addition of 12 mM MTT solution (10 μL; Bio-IDEA, Tehran, Iran) to each well, incubation was performed for 3 h at 37 °C. Next, the medium containing MTT was removed from the cells, and 100 μL of DMSO was added to each well and pipetted several times to form a completely uniform suspension. The solution was then shaken for 10 min at 37 °C until formazan crystals were completely dissolved. The obtained solution was added to a microtube and centrifuged in 1500 RPM for 5 min to eliminate any impurities. An ELISA reader was used to measure absorbance at 570 nm. For instance, all the mentioned steps were performed without adding the cells. The color concentration and percentage of cell survival were finally determined.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!