Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy
Gottschalk, E.; Czech, E.; Aksoy, B. A.; Aksoy, P.; Hammerbacher, J.
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Three-dimensional (3D) cell culture systems with tumor spheroids are being adopted for research on the antitumor activity of drug treatments and cytotoxic T cells. Analysis of the cytotoxic effect on 3D tumor cultures within a 3D scaffold, such as collagen, is challenging. Image-based approaches often use confocal microscopy, which greatly limits the sample size of tumor spheroids that can be assayed. We explored a system where tumor spheroids growing in a collagen gel within a microfluidics chip can be treated with drugs or co-cultured with T cells. We attempted to adapt the system to measure the death of cells in the tumor spheroids directly in the microfluidics chip via automated widefield fluorescence microscopy. We were able to successfully measure drug-induced cytotoxicity in tumor spheroids, but had difficulties extending the system to measure T cell-mediated tumor killing. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=193 SRC="FIGDIR/small/842039v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@c95725org.highwire.dtl.DTLVardef@785728org.highwire.dtl.DTLVardef@a23dceorg.highwire.dtl.DTLVardef@187bd74_HPS_FORMAT_FIGEXP M_FIG C_FIG
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