I want to measure the number of cells that died following addition of Tcells to the cancer cell spheroids with and without checkpoint inhibitors. I prefer a 96 well plate format. Since both the target and effectors are cells, and as there is a possibility of the Tcells dying itself, we cannot use standard viability assays. I am thinking of labelling the target cells with some dye that can be measured after its release in to the media following cell death. Is this a viable option?
how you can discriminate between apoptotic T cells and apoptotic tumor cells in suspention ? I think you should merge surface staining to apoposis assay. maybe at firstyou should stain T cells by specifc antibody (Anti CD3) to discriminate T cells from tumor cells, then evaluate apoptosis of tumor cells in CD3- population by 7AAD and AnnexinV . but the results reported by percentage...
Dear Dr Lucky,
could you try a clonogenic assay?
(Standard colony formation assays)
https://link.springer.com/article/10.1007%2Fs11060-019-03164-5
Dear Dr. Sasidharan,
You could try staining T cells and cancer cells from all treatment time points, and use zombie green flow cytometry dye for cell viability. Once the samples are stained with cell viability dye, you can carry out the cell viability analysis in a flow cytometer.
Hope this helps.
A simple AV/PI assay can be recommended as silver standard for cell death quantification.
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