A Three-Dimensional Co-culture Model of Infiltrating T Cells in Tumor Spheroids
Published: May 31, 2024
Abstract
Source: Lin, Y., et al. Monitoring Cancer Cell Invasion and T-Cell Cytotoxicity in 3D Culture. J. Vis. Exp. (2020).
This video demonstrates a method to establish a three-dimensional tumor spheroid and T cell co-culture. An agarose cast with microwells containing tumor spheroids is prepared, followed by the addition of T cells. After incubation, this setup allows studying the interaction of T cells with tumor spheroids in a controlled three-dimensional environment.
Protocol
1. Generation of spheroids Prepare and autoclave 2% agarose in 1x phosphate-buffered saline (PBS) (e.g., 1 g agarose in 50 mL of 1x PBS) and autoclave 35- and 81-microwell rubber molds. CAUTION: Avoid using low-melting agarose for generating the agarose casts for immunohistochemistry (IHC) processing. Prepare agarose casts NOTE: 35-microwell casts are used for invasion, immunofluorescence (IF) and cell isolation assays. 81-microwell ca…
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The authors have nothing to disclose.
Materials
| 3D Petri Dishes | Microtissues Inc | Z764019 & Z764051 | referred to as "rubber molds" in the protocols; 81-microwell & 35-microwell molds |
| 8-well Chamber Slides | Lab-Tek | 154534 | |
| Agarose Type I, low EEO | Sigma-Aldrich | A6013 | |
| DMEM, fetal bovine serum | ThermoFisher | 11965092, 16000044 | referred to as "cell culture medium" in the protocols |
| RPMI | ThermoFisher | 11875093 | for T-cell culture medium |
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