Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy

Abstract

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.

Activation of cytotoxic T lymphocytes by self-differentiated myeloid-derived dendritic cells for killing breast cancer cells expressing folate receptor alpha protein

“…We have further evaluated the cytotoxicity of T lymphocytes activated by SmartDC-FRα by using 3D spheroid culture, which better reflect the solid and heterogeneity nature of BC than monolayer culture ( Figure 6 ). The similarities of spheroid model to human tumors in terms of proliferative behavior and tumor microenvironment were reported [ 50 , 51 ]. The FRα-expressing MCF-7 and MDA-MB-231 spheroids were disrupt when cocultured with T lymphocytes activated by SmartDC-FRα ( Figure 6 ).…”