Establishment of CM for CLL. (A) Stromal NKTert cell line was cultured in complete media containing RPMI supplemented with 10% Fetal bovine serum (FBS) and penicillin-streptomycin (BM). Once the cells reached around 90% confluency, the supernatant was collected and filtered and stored at –80°C . Mononuclear cells, isolated from peripheral blood of samples from patients with CLL, were cultured in the presence of this CM (100%). More than 90% cells were CD19+ CD5+. (B) Relative CLL cells viability were measured when grown in presence of either CM, BM, or NKtert cells for 4 days (n = 5). CLL cells were separated from NKtert cells before measuring cell viability. (C) Annexin V staining of CLL cells after 2 days in culture with CM or BM (n = 4). Error bars represent standard error of the mean, P values were calculated using paired t test, and P value < .05 indicate statistical significance. Figure created with BioRender.com.