Protocol for analyzing immune cell infiltration in 3D pancreatic heterotypic spheroids composed of tumor cells and cancer-associated fibroblasts

STAR Protocols 2026 Open access

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Authors: Bessede Thomas, Vezzio-Vie Nadia, Freixinos Clara, Pirot Nelly, Bonnefoy Nathalie, Gongora Celine, Larbouret Christel, Gros Laurent
DOI: 10.1016/j.xpro.2026.104383
Cited by: 1
References: 10

Abstract

We present a protocol to generate and analyze 3D cancer models composed of tumor cells, cancer-associated fibroblasts (CAFs), and immune cells that mimic the complexity of the pancreatic ductal adenocarcinoma (PDAC) stroma. The protocol includes steps and conditions for establishing 3D co-cultures, embedding for histological analysis, and flow cytometry analysis for the immunophenotyping and characterization of tumor-infiltrating immune cells. These models are valuable for testing strategies and studying the roles of CAFs and immune cells in tumor growth, chemoresistance, and immunosuppression.

Fields of Study & Subjects

Cancer Cells and Metastasis Mathematical Biology Tumor Growth Pancreatic and Hepatic Oncology Research

Publication details

Journal / source
STAR Protocols
Publisher
Elsevier BV
Volume
7
Issue
1
Pages
104383–104383
Publication date
23 February 2026
Language
EN