Abstract

In vitro generation of dendritic cells (DCs) is advantageous for overcoming the low frequency of primary DCs and the difficulty of applying isolation techniques for studying DC immunobiology. The culture of bone marrow cells with granulocyte-macrophage colony-stimulating factor (GM-CSF) has been used extensively to generate bone marrow-derived dendritic cells (BMDCs). Studies have reported the heterogeneity of cells grown in murine GM-CSF culture based on the levels of MHCII expression. Although porcine DCs are generated by this classical method, the exact characteristics of the BMDC population have not yet been defined. In this study, we discriminated GM-CSF-grown BMDCs from gnotobiotic miniature pigs according to several criteria including morphology, phenotype, gene expression pattern and function. We showed that porcine BMDCs were heterogeneous cells that differentially expressed MHCII. MHCIIhigh cells displayed more representative of DC-like morphology and phenotype, including costimulatory molecules, as well as they showed a superior T cell priming capacity as compared to MHCIIlow cell. Our data showed that the difference in MHCIIhigh and MHCIIlow cell populations involved distinct maturation states rather than the presence of different cell types. Overall, characterization of porcine BMDC cultures provides important information about this widely used cellular model.

Highlights

  • Dendritic cells (DCs) are components of the immune system that can present antigens to T cells [1]

  • On the basis of these characteristics, we showed that non-adherent cells isolated from granulocyte-macrophage colonystimulating factor (GM-CSF)-grown Bone marrow cells (BMCs) cultures were heterogeneous in terms of their levels of MHCII expression

  • To generate bone marrow-derived dendritic cells (BMDCs) in vitro, BMCs were obtained from 3-week-old gnotobiotic miniature pigs and cultured with GM-CSF supplementation

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Summary

Introduction

Dendritic cells (DCs) are components of the immune system that can present antigens to T cells [1]. Conventional DCs (cDCs) provide signals for T cell activation and differentiation, and are regarded as professional antigen-presenting cells (APCs) of the immune system [2]. Study of these essential cells has been complicated by the low frequency of DC populations in blood and tissue. For this reason, the biology of DCs has been studied in cells grown in vitro from hematopoietic precursors, in the presence of growth factors [3].

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