The 26 chapters provide illustrated instructions and tips for avoiding common problems in reproducing clonogenic, colorimetric, fluorometric, and physiological assays in working with adherent/ nonadherent cell
After incubation for 30 min at 37[degrees]C, the medium was discarded and the cells were washed twice with PBS to remove the nonadherent cells
After a 1-h attachment period in the incubator, nonadherent cells
, mainly MCs, were separated using Percoll density gradient centrifugation at 2500 g for 15 min at 4[degrees]C, and cells that remained at the Percoll interface were aspirated and resuspended in PBS.
The culture medium was changed after 2 clays and nonadherent cells
Cells were allowed to adhere for 72h followed by the removal of nonadherent cells
and media were changed every 3 to 4 days.
After 72 hours, nonadherent cells
were discarded, and the adherent ones were thoroughly washed twice with DMEM.
2], washed three times with HBSS to remove nonadherent cells
, and equilibrated with DMEM that contained 10 per cent FBS before treatment.
After biofilm formation, the medium was aspirated and nonadherent cells
were removed by thoroughly washing the biofilm.
2] at 37 [degrees]C for 5-7 clays; the nonadherent cells
were then removed, whereas the plastic-adherent cells (the MSCs) were maintained under the same culture conditions.
were removed and adherent cells were cultured in 1 ml of fresh medium (5% FBS) containing 5 [micro]g/ml of E.
were removed 20 min after plating by changing the culture medium to DMEM/F12 containing 2% FBS.
for 20 min to remove nonadherent cells
and erythrocytes and eluted adherent cells using a combination of high shear (flow rate 40 [micro]L/min), calcium-free PBS, and air embolism.