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Assessing the capacity of IL-15 analogs (e.g. within TriKE molecules) to stimulate proliferation of NK-92 cells. NK-92 are deprived of cytokines overnight, then cultured with IL-15 analogs for two days. After this time, the expansion of viable cells is quantitatively assessed using a redox-sensitive dye that changes from blue/non-fluorescent in media to pink/fluorescent upon reduction in viable cells. It should be noted that NK-92 lack CD16, so delivery of IL-15 to these cells should not be enhanced by the anti-CD16 component of the TriKE as it would be for CD16+ NK-92 or CD16+ pNK cells. This protocol is adapted from the following assays: https://linkinghub.elsevier.com/retrieve/pii/0022175994903964 http://doi.wiley.com/10.1046/j.1440-1711.1998.00733.x |
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