2.3. Cell Cultures

The whole PBMC and isolated CLL cells cultures were cultured in a complete medium: RPMI 1640 with GlutaMAX (Gibco, France) supplemented with 10% of FBS (Life Technologies, France) and 100 µg/mL of penicillin/streptomycin (Sigma-Aldrich, France). Cells were plated at high density (10 × 10⁶ cell/mL) in tissue culture treated plates (Corning, USA), and maintained at 37 °C in a humidified atmosphere with 5% CO₂. In experiments to evaluate the NLC phenotype, cells were cultured in a 4 mL/well of 6-well plates, and to evaluate CLL cells viability, cells were cultured in a 200 µL/well of a 96-well plate or in a 400 µL/well of a 48-well plate in the case of fluorescence microscopy imaging.