Infinite f50 robotic absorbance microplate readers

To determine if CTEE affects cell viability, an MTT assay was used. In particular, pAECs were seeded in 96-well culture plates at a density of 3 × 10³ cells/well, incubated overnight; then, media were replaced with hESFM containing 5% FBS and increasing CTEE doses (10, 25, 50, and 100 μg/ml). After 24 h of incubation, MTT solution (5 mg/ml in PBS) was added to a final concentration of 0.5 mg/ml. After 2 h of additional incubation, 0.1 ml/well of MTT solubilization solution was added. Formazan Abs was determined at 570 nm, using Infinite® F50/Robotic absorbance microplate readers from TECAN Life Sciences (Männedorf, Switzerland).
The effect of CTEE on LPS-induced injury was also studied; pAECs were seeded in 96-well culture plates at a density of 12 × 10³ cells/well and incubated for 24 h, then exposed to different concentrations of CTEE (10, 25, 50, and 100 μg/ml) in the presence of LPS (25 μg/ml). After another 24 h of incubation, cell viability was evaluated by the MTT assay, as previously described.

pVW–MSCs were seeded in a 96-well plate (3 × 10³/well) and the day after, were treated with NaHS at increasing doses (0, 1, 5, 10, 50, 100, 200, 300 μM) for 24 h. The MTT assay was performed at the end of the treatment time following the manufacturer’s instructions. Briefly, the MTT substrate was added to the culture medium and incubated for 4 h, then the MTT solubilization solution was added to the cells to dissolve the formazan crystals. Formazan Abs were measured at 570 nm, using Infinite^® F50/Robotic absorbance microplate readers from TECAN Life Sciences (Männedorf, Switzerland). Cell morphology was routinely checked using an inverted Eclipse Microscope (TS100) equipped with a digital C-Mount Nikon phothocamer (TP3100)