Tissue homogenates from three control polyps, previously rinsed with sterile hydra medium, were plated on R2A agar and grown at 18 °C for three days (Fig. 4d).
Cells were collected by trypsinizaion 48 h after siRNA transfection, and 40,000 cells suspended in phenol red-free medium were plated on collagen I-coated 96 well plates.
104 cells/well in 90 μL of complete culture medium were plated in a 96-well plate and incubated for 24 h at 37 °C and 5% CO2, before adding 10 μL of either SLN or sorafenib in the culture medium.
The cultured glioma cells and co-cultures with hUCBSC (100 μL medium) were plated in 96-well microtiter plates at a final concentration of 0.5×105 cells/well.
Subsequently wells were washed three times with PBS and 2×105 CD4 positive mouse T cells in 200 μl culture medium were plated on each well.
Two thousand cells in 200 µl medium were plated into each well of 96-well plates and incubated overnight to allow cell adherence.
Lymph node cell suspensions in complete medium were plated at a concentration of 2×105 cells per well in 96-well plates in the presence of Ab2-β (5, 10 and 15 µg/mL) or 10 µg of gp43.
HUVECs were washing by serum free EBM medium twice and re-suspended containing 5×104 cells/well in 0.6 ml with medium were plated into 24-well inserts (Coring, 8 µm pore size) in duplicatation.
Three thousand cells (in 100 µl culture medium) were plated in a well of a 96-well clustered dish, and maintained until samples were taken at scheduled timepoints (48 120 h post-transfection).
MNCs (8×106 cells/ml medium) were plated on culture dishes coated with human fibronectin (Sigma) and maintained in endothelial basal medium (EBM; CellSystems) supplemented with 1 µg/ml hydrocortisone, 12 µg/ml bovine brain extract, 50 µg/ml gentamycin, 50 ng/ml amphotericin B, 10 ng/ml epidermal growth factor and 20% FCS.
The sorted cells suspended in serum-free medium were plated onto the Transwell inserts at 2.5 × 10 cells per well.
