MTT PBMC Proliferation and Cytotoxicity Assay

MTT cell proliferation and cytotoxicity assay kits (Beyotime Biotechnology, Shanghai, China) were used in this study for the cell viability assay of PBMCs stimulated by LPS and PHA [13 (link)]. Human PBMCs were resuspended to a concentration of 2 × 10⁴ living cells/mL. 100 μL aliquot was added immediately to each well of a 96 well flat bottom microplate in triplicate. LPS or PHA were then added to wells to final concentrations of 100 ng/ml (LPS) and 10 μg/ml (PHA). After 72 h incubation at 37 °C at 5% CO₂, PBMCs viability was assayed with an MTT kit. The ratio (test/control) of optical density at 570 nm was described as PBMCs-viability.