Our immune cell (IC) medium is a serum-free, ready-to-use cell culture medium for the in vitro cultivation and expansion of human immune cells. The medium has been specially optimized for the growth of T cells, CIK cells (cytokine-induced killer cells), LAK cells (lymphokine-activated killer cells) and NK cells (natural killer cells) in serum-free suspension cultures. Our IC medium was developed on the basis of RPMI 1640 and DMEM/F12 and enriched with special salts, trace elements, albumins, cholesterol, lipids and vitamins. It is ready-to-use and requires no further supplementation. Our IC medium deliberately does not contain any growth factors (such as recombinant cytokines) for cell differentiation or the enrichment of T cells from whole blood samples. Add these according to the individual requirements of your cells.
Our IC medium enables the proliferation of T cells from peripheral blood mononuclear cells (PBMCs)/whole blood, proliferation of NK, LAK and CIK cells after stimulation and the long-term cultivation of immortalized T cells. The serum-free cultivation of immune cells saves you time-consuming serum tests, prevents batch-to-batch variations and thus enables maximum reproducibility. It also reduces the risk of possible contamination.
Peripheral blood cells are arrested in the G0 phase and must be stimulated with mitogens (plant lectins, e.g. phytohemaggluthinin PHA; or antibodies such as anti-CD3 ) in order to be cultivated in vitro.
Tips for the stimulation of T cells:
T cell density of 1x105/ ml
Addition of PHA according to manufacturer's instructions (1-5 μg/ml)
Incubation time is between 48-72 hours (depending on the type and origin of the lymphocytes and their use)
Addition of fresh IZ medium and further stimulation of the T cells is necessary to continue the culture
Please note: The proliferation rate of primary lymphocytes from whole blood is limited
Tips for stimulation/cultivation of CIK cells:
CIK are enriched by adding IFN-γ, mAb anti-CD3 (OKT3) and IL-2 to isolated PBMCs (peripheral blood mononuclear cells). After 2-3 weeks in vitro culture, their concentration is increased 1000-fold.
Application
Proliferation of T cells from peripheral blood mononuclear cells (PBMCs)