Culture media

Historically, mononuclear blood cells have always been cultured in vitro in a medium (usually RPMI 1640 or IMDM) containing serum as a source of undefined growth supporter. FBS and pooled human AB serum are routinely used in this system. However, cells intended for clinical use should preferably not be cultured with animal or human sera due to possible viral transmissions and side-effects, including patients' anaphylactic reactions caused by the presence of xenogeneic/allogeneic proteins remaining on the cell surface. Therefore, when possible, serum-free culture media should be used for cells intended for patient reinfusion. In response to this need, different companies have developed serum-free media (Table 1) specifically designed to support the in vitro growth of T cells, such as LAK cells and TILs, or hematopoietic progenitors, such as CD344 cells. In addition, the media used for in vitro expansion of blood-derived cells for reinjection into humans must be endotoxin-free (endotoxin units (EU) <0.05 per ml) and, if possible, antibiotic-free. When the addition of antibiotics is required, preference is to be given to antibiotics for which the risk of allergic reactions is low (such as gentamycin sulfate). Consequently, p-lactamic antibiotics (e.g. penicillin) will not be used.

Growth and/or differentiation factors (such as GM-CSF, G-CSF, IL-3, stem cell factor (SCF) for hematopoietic progenitors, IL-2 for LAK cells and TILs, IL-4, GM-CSF and tumor necrosis factor a (TNFa) for dendritic cells) are usually added to the culture media to support the growth and/or potentiate the biological functions of these blood-derived cells. Only commercially available and purified recombinant human growth factors can be used in clinical-grade cultures. Conditioned media containing 'growth factor activities' cannot be used to grow cells for use in humans.

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