Celltiter 96 aqueous one solution proliferation assay

Cell viability of macrophages, pre-adipocytes, and mature adipocytes treated with extracts (31–500 μg/mL) was performed with the CellTiter^® 96 Aqueous One Solution Proliferation assay (Promega Corporation, Madison, WI, USA) following manufacturer’s instructions.

Viability was assessed with MTS reagent, CellTiter 96 AQueous One Solution Proliferation Assay from Promega Corporation (Madison, WI, USA) or by Guava ViaCount flow cytometer assay (Millipore, Burlington, MA, USA).

Cell viability after viral infection was measured using a colorimetric cell lysis test (MTS). Ten thousand cells per well were plated on 96-well plates in growth medium (GM) supplemented with 5% fetal calf serum (FCS). Confluent cell layers were infected in triplicates with different concentrations of virus for 1 hour in 2% FCS GM. After that, 5% FCS GM was added into wells and cells were incubated until cell viability was measured by MTS according to manufacturer’s instructions (Cell Titer 96 AQ_ueous One Solution Proliferation Assay, Promega, Madison, WI). Inactivated vaccinia virus was used as a non-replicating control, inactivation of the virus was done as described previously [11 (link)].

To determine viability, IEC-6 and HepG2 cells were grown in 96-well plates at 5.0 × 10⁵ cells per mL. After 24 h, the cells were treated with the digested CSF and the CSE (200 μg mL⁻¹) and incubated for 24 h. Cell viability was determined using the CellTiter^® 96 Aqueous One Solution Proliferation assay (Promega, Madison, WI, USA) as indicated by the manufacturer.

The cell viability of hepatocytes treated as indicated in Section Cell Culture Model of NAFLD Prevention was measured colorimetrically using the CellTiter 96 Aqueous One Solution Proliferation assay (Promega Corporation, Madison, WI, USA).

Growth curves and cellular morphology were assessed using Incucyte (Essen Instruments, Ann Arbor, MI), a kinetic live cell imaging system. Proliferation was measured through quantitative kinetic processing metrics derived from time-lapse image acquisition and presented as percentage of culture confluence over time. Effect of treatment on cell viability was quantified using Celltiter⁹⁶ Aqueous One Solution Proliferation Assay (Promega, Madison, WI).

Influence of matriptase inhibitors in phenol red free Williams medium E on the viability of HepK2 and HepK6 cocultures was tested. Monolayer cocultures, grown on a 96-well plate for 24 h, were incubated with matriptase inhibitors for 24 h in treated groups. The control cells were incubated only with phenol red free Williams medium E. After removal of the medium and washing the cells 3-fold with phosphate-buffered saline (PBS), 20 μL of CellTiter 96 aqueous one solution proliferation assay (MTS, Promega, Bioscience, Budapest, Hungary) containing tetrazolium compound and an electron coupling reagent, phenazine ethosulfate, was pipetted into each well of the 96-well assay plate containing the samples in 100 μL of culture medium. The plate was incubated with dye for 2 h. Viability of HepK2 and HepK6 cocultures was measured at 490 nm using EZ Read Biochrom 400 microplate reader.