Protocol for co-culture of murine bone marrow-derived dendritic cells with OT-I mouse CD8+ T cells to evaluate immune activation efficacy

Jiang, Min; Matzek, Dana; Nguyen, Anny; Popper, Bastian; Merkel, Olivia M. (2026): Protocol for co-culture of murine bone marrow-derived dendritic cells with OT-I mouse CD8+ T cells to evaluate immune activation efficacy. STAR Protocols, 7 (2): 104621. ISSN 26661667

Abstract

Highlights

• Steps for isolating bone marrow using centrifugation-based approach
• Protocol for generating BMDCs from murine bone marrow
•Instructions for isolating OT-I CD8+ T cell by magnetic separation
• Guidance on co-culture and functional readouts of CD8+ T cell activation

Summary

Preliminary evaluation of vaccine-induced immune activation can be performed with minimal animal use, in line with the principles of the 3Rs (replacement, reduction, and refinement) in animal experimentation. Here, we present a protocol to assess murine antigen-specific CD8+ T cell activation using a co-culture model of bone marrow-derived dendritic cells (BMDCs) and OT-I transgenic CD8+ T cells. We describe the steps for transfecting BMDCs with antigen-encoding mRNA formulations and assessing T cell responses after co-culture by intracellular cytokine staining and proliferation.

• Dokumententyp: Artikel (LMU)
• Organisationseinheit (Fakultäten): 18 Chemie und Pharmazie > Department für Pharmazie - Zentrum für Pharmaforschung
• DFG-Fachsystematik der Wissenschaftsbereiche: Naturwissenschaften
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 491502892