MTT Assay solution

Protocol tips

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	After 24 hours, the culture medium was aspirated and the cells were rinsed twice with PBS (Roche). The MTT solution was prepared by adding 8 mg of MTT to 10 mL fresh DMEM and 500 μL of MTT solution was added to each well. Four hours later, the MTT solution was removed and each well was incubated with 500 μL isopropanol for 30 min.	The extinction values of the isopropanol-diluted formazan were measured at 570 nm wavelength. The viability index was determined by calculating the ratio of the extinction values of labeled cells compared to unlabeled control cells.

Protocol tips
After 24 hours, the culture medium was aspirated and the cells were rinsed twice with PBS (Roche). The MTT solution was prepared by adding 8 mg of MTT to 10 mL fresh DMEM and 500 μL of MTT solution was added to each well. Four hours later, the MTT solution was removed and each well was incubated with 500 μL isopropanol for 30 min.
Downstream tips
The extinction values of the isopropanol-diluted formazan were measured at 570 nm wavelength. The viability index was determined by calculating the ratio of the extinction values of labeled cells compared to unlabeled control cells.

Publication protocol

Second, a 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan tetrazolium bromide assay (MTT assay, Sigma-Aldrich) was performed at days 3, 7, 14, and 28. This technique provides an indirect measure for the number of proliferating cells. Here, 104 cells/well were plated in 24-well plates (BD Falcon, Heidelberg, Germany) in ASC medium. After 24 hours, the culture medium was aspirated and the cells were rinsed twice with PBS (Roche). The MTT solution was prepared by adding 8 mg of MTT to 10 mL fresh DMEM and 500 μL of MTT solution was added to each well. Four hours later, the MTT solution was removed and each well was incubated with 500 μL isopropanol for 30 min. The extinction values of the isopropanol-diluted formazan were measured at 570 nm wavelength. The viability index was determined by calculating the ratio of the extinction values of labeled cells compared to unlabeled control cells.

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