Effect of culture and maturation on human monocyte-derived dendritic cell surface markers, necrosis and antigen binding

Abstract

Dendritic cells (DC) are antigen-presenting cells (APC) that are important for innate and acquired immune responses. Owing to their involvement in autoinflammation, autoimmunity and cancer, DC are useful cellular models for biomedical research. Appropriate DC production in vitro could aid the study of DC in many human diseases. We used fluorochrome-based flow cytometry assays to analyze the effects of culture period and maturation of monocyte-derived DC (MoDC) on their viability and necrosis, purity, CD11c expression and phagocytic capacity. The morphological changes that occur as purified monocytes become DC were assessed at 24 and 72 h, and 6 and 9 days in culture. The dynamics of certain cell surface markers of monocytes and mature MoDC (mMoDC) also were assessed using fluorescence-based assays. We found that day 6 of culture yielded the most functional immature MoDC (iMoDC) with maximal viability, purity, CD11c expression and appropriate phagocytic capacity. Mass production of viable MoDC could be useful for immunotherapy.

Key words::

• antigen-presenting cells
• CD11c
• CD14
• cell culture
• dendritic cells
• flow cytometry
• fluorescence
• monocyte-derived dendritic cells
• necrosis
• phagocytosis

Acknowledgments

We thank Mr. Abbas Bahari, Ms Nafiseh Tabasi and Ms. Shahnaz Zamani Taghizadeh Rabe for their technical help. This work was supported financially by a grant from Ferdowsi University of Mashhad, Iran.

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this paper.