Cytotoxicity Assay of Immune Cells

Briefly, BNL or 4 T1 cells were seeded into a 96-well plate and allowed to attach for 1 h. Subsequently, suspension of splenocytes derived from mice in different groups was added at a ratio of splenocytes to target (BNL or 4 T1) cells varying from 2:1, 10:1 to 20:1, respectively, and incubated with BNL cells for 4 h. Lactate dehydrogenase (LDH) release from the lysed BNL cells was measured as an indicator of cytotoxic activity with Cytotoxicity Detection Kit^PLUS (LDH) (Sigma-Aldrich, #4744926001) [24 (link), 43 (link)]. All experiments were performed in triplicate.

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Bai X., Zhou Y., Yokota Y., Matsumoto Y., Zhai B., Maarouf N., Hayashi H., Carlson R., Zhang S., Sousa A., Sun B., Ghanbari H., Dong X, & Wands J.R. (2022). Adaptive antitumor immune response stimulated by bio-nanoparticle based vaccine and checkpoint blockade. Journal of Experimental & Clinical Cancer Research : CR, 41, 132.

Publication 2022

Cytotoxicity Detection KitPLUS (LDH)

Cells Cytotoxicity Lactate dehydrogenase Mice

Corresponding Organization :

Other organizations : Brown University, Providence College, Rhode Island Hospital, First Affiliated Hospital of Harbin Medical University, Harbin Medical University

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Variable analysis

independent variables

Ratio of splenocytes to target (BNL or 4T1) cells (2:1, 10:1, 20:1)

dependent variables

Lactate dehydrogenase (LDH) release from the lysed BNL cells (as an indicator of cytotoxic activity)

control variables

BNL or 4T1 cells seeded into a 96-well plate and allowed to attach for 1 hour
Incubation time of splenocytes with BNL cells (4 hours)

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