Begin with a culture plate containing an agarose cast. The cast comprises a seeding chamber with multiple microwells.
Each microwell contains a tumor spheroid in a culture medium. The spheroids are three-dimensional aggregates of tumor cells.
First, remove the medium surrounding the agarose cast. Then, remove the medium within the cast from one corner of the seeding chamber to avoid the loss of spheroids.
Introduce a suspension of tumor-specific T cells dropwise into the seeding chamber, allowing the cells to settle into the microwells containing the spheroids.
Next, add a culture medium around the agarose cast and incubate the plate under physiological conditions to develop a co-culture.
During incubation, the T cells surround the spheroid. A subset of T cells infiltrate the spheroid, while others remain in the periphery, establishing a three-dimensional co-culture model.
Related Videos
Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity
Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses
Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells
ABOUT JoVE
Copyright © 2024 MyJoVE Corporation. All rights reserved