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Image-based 3D cell culture cytotoxicity assay v1
Elinor Gottschalk, B. Arman Aksoy, Pinar Aksoy, Eric Czech, Jeff Hammerbacher
GUIDELINES This protocol describes an imaged-based 3-dimensional (3D) cell culture cytotoxicity assay using multicellular tumor spheroids (in this case MC38 cells) sensitized with hgp100 peptide and killing them with pmel-1 T cells or the apoptosis-…
GUIDELINES This protocol describes an imaged-based 3-dimensional (3D) cell culture cytotoxicity assay using multicellular tumor spheroids (in this case MC38 cells) sensitized with hgp100 peptide and killing them with pmel-1 T cells or the apoptosis-inducing antibiotic, Staurosporine. Our goal was to explore an in vitro 3D cell culture system to study T cell-mediated cytotoxicity as a potential way to more rapidly and relevantly test the effect of T cell manipulation on T cell cytotoxicity before moving to the mouse model. In this setup, the T cells are required to migrate towards the tumor spheroids that are already suspended in a gel matrix, there is minimal manipulation of the samples post-coculture because the samples are imaged directly in the chips to measure cell death, and there is somewhat higher throughput than confocal imaging provides (more spheroids being imaged per sample via widefield microscopy). We used 3D cell culture chips from AIM Biotech (Singapore) and adapted protocols published by: AIM Biotech:
Published in
protocols.io
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0
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0 | 2019 |
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