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Protocol for analyzing immune cell infiltration in 3D pancreatic heterotypic spheroids composed of tumor cells and cancer-associated fibroblasts
Article de revue scientifique   Open Access   Avec comité de lecture

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

Bessede Thomas, Vezzio-Vie Nadia, Freixinos Clara, Pirot Nelly, Bonnefoy Nathalie, Gongora Celine, Larbouret Christel et Gros Laurent
STAR Protocols, Vol.7(1)
03/2026
PMCID: PMC13080457
PMID: 41734060

Résumé

Cancer Cell Biology Immunology Humans Cancer-Associated Fibroblasts Spheroids, Cellular Pancreatic Neoplasms Coculture Techniques Carcinoma, Pancreatic Ductal Cell Line, Tumor Flow Cytometry
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.

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