The inoculum was then replaced by fresh medium and cells were further incubated.
After four weeks, the culture medium was replaced with the serum-free growth medium and cells were serum-starved for 72 hours before treatment.
Then, the culture medium was replaced with 10% FBS supplemented DMEM medium and cells were irradiated using a homemade device at the desired light dose (10 or 27 J/cm27.
18 24 hrs after plating passaging medium was changed to complete culture medium and cells were grown for either 48 or 24 hrs prior to live cell imaging or fixation and immunocytochemistry procedures respectively.
Using the pharmacokinetic model, the cumulated activity concentrations in the medium and cells were calculated.
Also, a first-order pharmacokinetic model was used to simulate the closed compartmental system between the medium and cells.
Briefly, for the Click-iT method (Invitrogen), EU, a nucleotide analog, was added into the medium and cells were incubated for 1 h at 37°C.
Then, the cell medium was replaced by a serum-free medium and cells were incubated for 30 min at 37 °C under 5%% CO2 and 95%% humidity.
Experimental data were then fitted to this model and used to estimate the transfer coefficients between medium and cells, k(m)(c), and between cells and medium, k(c)(m).
The medium and cells were collected after 48 hours.
Medium and cells were harvested, and immunoprecipitated again apoA-I, apoA-IV, and albumin antibodies.
