Comparison of different media to expand t cells over a time period of 20 days. X-vivo 15 , TexMacs and cell genix were compared and checked for viability, identity and fold increase. No FBS, HS or HSA has been added
CIK (CD3+CD56+) generation: 30×10exp6 of cells were seeded in 15 ml of culture medium. We cultured the CIKs in T75 fasks (Corning Incorporated, NY, USA).
At Day 0, 1000 U/ml IFN-? was added (Boehringer Ingelheim, Vienna, Austria).
At Day 1, 50 ng/ml CD3 Pure, which is IgG anti-CD3 (OKT-3) (170-076-124, Miltenyi Biotec, Bergisch Gladbach, Germany), and 300 IU/ml of Proleukin which is IL-2 cytokines (Novartis, Origgio, VA, Italy) were added .
Fresh medium and Proleukin (300 U/ml) were added weekly (every 3 days) during culture, and the cell concentration was maintained at 1-1.5×106 cells . The fold increase of CIKs was calculated as follow: (%CD3+×total cells counted at Day 20–21)/(%CD3+×seeded cells at Day 0). After 20–21 days, CIK cells culture was stopped and the quality controls were performed (cell count and viability, immunophenotype and sterility).