Iranian Journal of Immunology

Efficient Generation of Bone Marrow-Derived Murine Dendritic Cells: A Protocol ‎Optimization Study

Document Type : Original Article

Authors

Marziyeh Mousazadeh 1
Maryam Nikkhah 1
Negar Seyed 2
Sima Rafati 2
Saman Hosseinkhani 3

1 Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran‎.

2 Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran.

3 Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.‎

https://doi.org/10.22034/iji.2026.108143.3086
Abstract
Background: Dendritic cells (DCs) are the most potent antigen-presenting cells, playing a central role in T cell activation and the stimulation of B cell antibody production.  Owing to their immunological significance, DCs have been extensively explored for applications in cancer immunotherapy, dendritic cell–based vaccines, and strategies to overcome the immunosuppressive tumor microenvironment.
Objective: Given the broad applications of DCs and the need for efficient generation protocols, the present study aimed to optimize key parameters influencing the production of bone marrow-derived dendritic cells (BMDCs).
Methods: Variables such as mouse strain, cytokine concentrations, culture plate type, differentiation duration, incubation temperature, cell seeding density, and media replacement intervals were evaluated. CD11c expression was used as the primary marker to quantify BMDCs, while CD80 and CD86 were used as indicators of maturation status.
Results: The optimized protocol yielded 60-70% CD11c+ BMDCs and 70-80% CD80/CD86 expression within 5 days.
Conclusion: This robust and highly reproducible protocol offers valuable applications for in vitro dendritic cell generation studies.

Keywords

Bone marrow-derived dendritic cells
Cell isolation protocols
CD11c
Cell differentiation
‎Granulocyte-macrophage stimulating factor‎
Iranian Journal of Immunology
Volume 23, Issue 1
Winter 2026
Pages 16-33

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