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Updated: Sep 13, 2025

A Macrophage-Tumor Spheroid Co-Invasion Assay
Published on: January 24, 2025
Protocol for analyzing T cell and macrophage motility in a mouse pancreatic cancer spheroid model using light-sheet
Vanessa Zimmer1, Benedikt Slusny1, Katrin Roth2
1Department of Gastroenterology, Endocrinology, Infectious Diseases and Metabolism, Philipps University Marburg, 35043 Marburg, Germany.
Abstract:
The interaction of macrophages and T cells is pivotal for shaping the immune response to cancer. Here, we present a protocol for investigating the co-culture of macrophages and T cells in a pancreatic cancer spheroid model. We describe steps for analyzing migration patterns by light-sheet microscopy, generating murine bone marrow-derived macrophages (BMDMs) and cytotoxic T lymphocytes (CTLs), and staining CTLs for image analysis. We then detail procedures for generating spheroids using a mouse pancreatic cancer cell line. For complete details on the use and execution of this protocol, please refer to Slusny et al.1.

