ArticleJournal of nanobiotechnology2022
3D hanging spheroid plate for high-throughput CAR T cell cytotoxicity assay.
Article in Journal of nanobiotechnology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Microfluidic Cytotongue system enables early, label-free detection of CAR-T-induced mechanical softening in tumor spheroids.Microsystems & nanoengineering · 2026Article
- Advantages and research progress of three-dimensional culture systems for lung cancer drug screening (Review).Oncology letters · 2026Review
- CAR-T cells directed toward PD-L1 demonstrate potent, antigen-specific activity against cholangiocarcinoma: A proof of concept study.Molecular therapy. Oncology · 2026Article
- Review
- Article
- Advances in Cytotoxicity Testing: From In Vitro Assays to In Silico Models.International journal of molecular sciences · 2025Review
- A 3D tumor spheroid model with robust T cell infiltration for evaluating immune cell engagers.iScience · 2025Article
- An ovalbumin-based hydrogel loaded with dendrobium polysaccharide for promoting wound healing while reducing inflammations.Scientific reports · 2025Article
- Leveraging microphysiological systems to expedite understanding of host-parasite interactions.PLoS pathogens · 2025Review
- Dual-parameter tomographic imaging of attenuation and backscattering coefficients for quantitative evaluation of immune cell-mediated cytotoxicity in tumor spheroids.Theranostics · 2025Article
- From spheroids to organoids: next-generation models for CAR-T cell therapy research in solid tumors.Frontiers in immunology · 2025Review
- Recapitulating Glioma Stem Cell Niches Using 3D Spheroid Models for Glioblastoma Research.Biosensors · 2024Review
- Imaging-Based Efficacy Evaluation of Cancer Immunotherapy in Engineered Tumor Platforms and Tumor Organoids.Advanced healthcare materials · 2024Review
- Three-dimensional breast cancer tumor models based on natural hydrogels: a review.Journal of Zhejiang University. Science. B · 2024Review
- CAR T cell infiltration and cytotoxic killing within the core of 3D breast cancer spheroids under the control of antigen sensing in microwell arrays.APL bioengineering · 2024Article
- Reshaping the tumor immune microenvironment to improve CAR-T cell-based cancer immunotherapy.Molecular cancer · 2024Review
- Tumoroids, a valid preclinical screening platform for monitoring cancer angiogenesis.Stem cell research & therapy · 2024Review
- Advancements in 3D In Vitro Models for Colorectal Cancer.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Review
- Networked Cluster Formation via Trigonal Lipid Modules for Augmented Ex Vivo NK Cell Priming.International journal of molecular sciences · 2024Article
- Three-Dimensional Bioconjugated Liquid-Like Solid (LLS) Enhance Characterization of Solid Tumor - Chimeric Antigen Receptor T cell interactions.bioRxiv : the preprint server for biology · 2023Article
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8 authors.
Funding
Abstract
backgroundMost high-throughput screening (HTS) systems studying the cytotoxic effect of chimeric antigen receptor (CAR) T cells on tumor cells rely on two-dimensional cell culture that does not recapitulate the tumor microenvironment (TME). Tumor spheroids, however, can recapitulate the TME and have been used for cytotoxicity assays of CAR T cells. But a major obstacle to the use of tumor spheroids for cytotoxicity assays is the difficulty in separating unbound CAR T and dead tumor cells from spheroids. Here, we present a three-dimensional hanging spheroid plate (3DHSP), which facilitates the formation of spheroids and the separation of unbound and dead cells from spheroids during cytotoxicity assays.
resultsThe 3DHSP is a 24-well plate, with each well composed of a hanging dripper, spheroid wells, and waste wells. In the dripper, a tumor spheroid was formed and mixed with CAR T cells. In the 3DHSP, droplets containing the spheroids were deposited into the spheroid separation well, where unbound and dead T and tumor cells were separated from the spheroid through a gap into the waste well by tilting the 3DHSP by more than 20°. Human epidermal growth factor receptor 2 (HER2)-positive tumor cells (BT474 and SKOV3) formed spheroids of approximately 300-350 μm in diameter after 2 days in the 3DHSP. The cytotoxic effects of T cells engineered to express CAR recognizing HER2 (HER2-CAR T cells) on these spheroids were directly measured by optical imaging, without the use of live/dead fluorescent staining of the cells. Our results suggest that the 3DHSP could be incorporated into a HTS system to screen for CARs that enable T cells to kill spheroids formed from a specific tumor type with high efficacy or for spheroids consisting of tumor types that can be killed efficiently by T cells bearing a specific CAR.
conclusionsThe results suggest that the 3DHSP could be incorporated into a HTS system for the cytotoxic effects of CAR T cells on tumor spheroids.
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