3D bioprinting technology developed for cancer immunotherapy
]. The hydrogel was incubated for 30 min and centrifuged to obtain 3D cultured cells. The cytotoxicity was investigated using a calcein-AM-based assay. Target cancer cells were stained with calcein-AM for 1 h at 37 °C. The labeled target cells and NK92 cells were placed in 96-well round-bottom plates at effector-to-target ratios of 2:1 and 1:1 for 4 h. The maximum release was measured by adding 4% Triton X-100 to the target cell.
Cytokine release, which affects the target, was measured using an ELISA. The supernatant of the tumor cell media co-cultured with NK cells was collected, and the concentration of released cytokines was investigated. An ELISA kit was used according to the instructions of the manufacturer.The cells were washed with FACS buffer and centrifuged to obtain cell pellets. The cell pellets were stained with antibodies for 30 min and analyzed by fluorescence-activated cell sorting .
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