Turner biosystems 20 20n luminometer

In order to assess viability of cells grown on the various mineralized chitosan fiber mats, the Promega Cell Titer Glo assay (Promega, Madison, WI) was utilized to qualitatively measure the amount of adenosine triphosphate (ATP) in culture. Both chitosan-genipin and chitosan-HDACS mats were cut into squares (about 1.4 cm × 1.4 cm), covering approximately 90% of the 2.0 cm² area of each well. Cells were seeded at a density of 5 × 10⁴ cells/well and grown in 500 µL of the supplemented α-MEM media. The control was MLO-A5 cells seeded at the same density into the tissue culture polystyrene (TCP) wells. Media was changed every three days during this experiment. After each time point, the assay was performed according to the Promega Protocol. Briefly, the luminescent reagent was mixed with the contents of the well and shaken for 2 min to induce cell lysis. After allowing the signal to stabilize for 10 min, the entire contents of the wells were placed into a Turner Biosystems 20/20ⁿ Luminometer (Promega, Madison, WI) and scanned using the preloaded Promega Cell Titer Glo protocol. Measurements for each well were recorded three times, with the reported values being the average with standard deviation.