Cell viability was assessed using the MTT Cell Proliferation and Cytotoxicity Detection Kit (Beyotime). For the colony formation assay, cells were seeded into 6-well plates at a concentration of 1×103 cells per well and incubated at 37 °C with a 5% CO2 atmosphere. After two weeks, cells were fixed and stained with Giemsa stain, modified solution (Sigma). Colonies containing more than 50 individual cells were counted.
Mtt cell proliferation and cytotoxicity detection kit
Manufactured by Beyotime
The MTT Cell Proliferation and Cytotoxicity Detection Kit is a colorimetric assay used to measure cell proliferation and cytotoxicity. It utilizes the yellow tetrazolium salt MTT, which is reduced by metabolically active cells to form purple formazan crystals. The amount of formazan produced is directly proportional to the number of viable cells, and can be quantified by measuring the absorbance.
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2 protocols using mtt cell proliferation and cytotoxicity detection kit
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Cell Viability and Colony Formation
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MTT Assay for Cell Proliferation
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Neuro2a cells were seeded into a 96-well plate at a density of 10^5. After overnight culture, transfection or Butylphthalide treatment was given according to the requirements of each group with MTT cell proliferation and cytotoxicity detection kit (Beyotime). Then 10 μL MTT solution was added to each well and they were continued to be incubated for 4 hours in the cell incubator. 100 μL formazan solution was added into each well. After mixing, they were continued to be incubated in the cell incubator. The incubation did not stop until the formazan was completely dissolved under the ordinary optical microscope. Generally, after incubation at 37°C for 3–4 hours, all purple crystals will dissolve. The absorbance value was measured at 570 nm and observed under 20x optical microscope.
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