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. 2016 Jan 27;6:19772. doi: 10.1038/srep19772

Figure 6. Assessment of the ADCC bioassay using frozen NK cell lines and human PBMCs.

Figure 6

(a,b) BT-474 serving as target cells were incubated with frozen NK92MI + CD16a serving as effector cells at an E:T ratio of 4:1 without (white circles) or with Trastuzumab (black circles). ADCC activity was then detected using the flowcytometric (a) or the calcein-release (b) assay. (c) The stability of frozen human PBMCs was assessed using the flowcytometric assay. PBMCs were isolated and cryopreserved at –80˚C using CellBanker I. Portions of the PBMCs were then thawed every 3 days after initially being frozen. ADCC activity was detected, using BT-474 as target cells at an E:T ratio of 4:1, without (white circles) or with 10 μg/mL of Trastuzumab (black circles). The results of the analyses of dead target cells (%) are presented as mean values ± SD. CV values were calculated using the following formula: SD/mean×100 (%).