Cell and gene therapies have shown great promise in treating hematologic cancers, however, effectively applying these treatment modalities to solid tumors presents unique challenges. Emerging strategies and novel approaches, including the use of three-dimensional (3D) cell models and advanced quantitative imaging techniques, are providing new insights to guide this area of research and overcome these challenges.
In vitro 3D models help bridge the gap between conventional two-dimensional in vitro testing and animal models. 3D cell cultures possess several of the same features present in tumors, such as complex cell-cell and cell-matrix interactions. By mimicking a 3D microenvironment with extracellular matrices (ECM), 3D tumor spheroids resemble in vivo tissue and offer improved models for hypoxia, central necrosis, drug penetration, in addition to the response, resistance, production, and deposition of the extracellular matrix.
Agilent Cell Analysis instrumentation and application solutions provide robust, quantitative methods for monitoring immune cell-induced inhibition of 3D tumor spheroid growth and invasion. Automated sample maintenance, image acquisition, and analysis offers a simple and powerful approach that supports long-term immuno-oncology applications and high-throughput studies.
Explore Agilent cell analysis solutions to learn about comprehensive real-time 3D tumor spheroid assays that provide:
Adoptive cell therapies such as chimeric antigen receptor (CAR) T lymphocytes have ushered in a new paradigm of cancer immunotherapy. In this study, we utilize both widefield and confocal capabilities of the Cytation C10 to study T lymphocytes genetically modified to express the variable chain fragment specific for the epithelial cell adhesion molecule (EpCAM), also known as CD326, on their surface.
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Pre-activated PBMCs killing of HT-1080 spheroids (A). HT-1080 cells labelled by eLenti red (2.5K/well) were seeded into ULA plate and spheroids were formed. Pre-activated PBMCs (25K/well) or non-activated PBMCs (25K/well) were added into the wells 3 days post cell seeding. 2x2 stitched images were taken once per 4 hours over 10 days using 10x objected. Images show brightfield and red nuclei of HT-1080 cells on days 0, 2, 4 and 6 after treatment with pre-activated PBMCs at ratios of 2.5:1, 5:1, 10:1, and 20:1.
E:T ratio-dependent killing of HT1080 spheroids by the pre-activated PBMCs. Pre-activated PBMCs killing of HT-1080 spheroids. HT-1080 cells labelled by eLenti red (2.5K/well) were seeded into ULA plate and the spheroids were formed. Different counts of pre-activated or non-activated PBMCs were added three days later to achieve different E:T ratios. Images show brightfield and red nuclei of HT-1080 cells on days 0, 2, 4 and 6 after treatment with pre-activated PBMCs at different E:T ratios. E:T ratio-dependent killing of HT-1080 spheroids by pre-activated PBMCs (B) or non-activated PBMCs (C) was measured via red total integrated intensity.
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At a ratio of 10:1, Pre-activated PBMCs show a strong killing effect compared to Non-Activated PBMCs by live cell video. Data was quantified using red count (not shown) and red total integrated intensity; and compared to 2D assays.
The Agilent BioTek Cytation cell imaging multimode reader, in conjunction with Agilent BioTek BioSpa 8 automated incubator is well suited to the long-term kinetic cytotoxicity assays using 3D technologies and immune cells. In this featured application note, NK cell-mediated cytotoxicity assays were conducted using a 3D collagen-based hydrogel and spheroids, respectively.
In order to evaluate and optimize adoptive T cell immunotherapies, sensitive in vitro methods must be included in the testing process. In the procedure described here, phenotypic and quantitative assessments of 2D and 3D target cell necrotic induction were made using automated live cell imaging. It was found that direct activation of T cells produced a significantly greater cytotoxic effect than general activation suggesting that T cells can be "taught" to target and destroy specific target cells.
In the first of this two-part educational webinar series on 3D cell culture, we’ll present: an overview of available 3D culture formats and reagents, essential considerations for setting up and optimizing 3D models and tips for effectively conducting imaging-based 3D cell culture assays.
In the second half of this webinar series on 3D cell culture we’ll build on the concepts covered in part one and present: tips for effectively conducting quantitative analysis on spheroids, instruments and hardware features optimized for 3D cell culture samples and the experimental approach and analysis methods used in a variety of 3D cell culture applications, including cell proliferation and viability, migration and invasion, quantification of lipid droplet within a hepatic model, characterization of intestinal organoid morphology, and stem cell differentiation.
Comprehensive overview of cell health and viability applications that provide multi-parametric real-time automated readouts using simultaneous cellular impedance and live cell imaging on xCELLigence RTCA eSight.
In one instrument, capture sensitive changes in cell number, morphology, and attachment by impedance monitoring and simultaneously visualize them by live cell imaging. Download the xCELLigence RTCA eSight Live Cell Analysis eBook to learn more.
Interested in learning more? Request a demo by clicking on the link below.
For Research Use Only. Not for use in diagnostic procedures.
