Type of monocyte immunomagnetic separation affects the morphology of monocyte-derived dendritic cells, as investigated by scanning electron microscopy

• Published in: Journal of immunological methods Vol. 439; pp. 79 - 82
• Main Authors: Kowalewicz-Kulbat, M., Ograczyk, E., Krawczyk, K., Rudnicka, W., Fol, M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-12-2016
• Subjects: Adult; Antibodies, Monoclonal - immunology; Antigens, CD - immunology; Cell Shape - drug effects; Cell Size - drug effects; Cells, Cultured; Dendritic cells; Dendritic Cells - drug effects; Dendritic Cells - immunology; Dendritic Cells - ultrastructure; Glycophorin - immunology; Humans; Immunomagnetic Separation - methods; Lipopolysaccharides - pharmacology; Magnetic-activated cell sorting; Microscopy, Electron, Scanning; Monocytes - drug effects; Monocytes - immunology; Monocytes - ultrastructure; Scanning electron microscopy; PBS; Scanning electron microscopy; MoDCs; IL; Dendritic cells; MACS; PBMC; Magnetic-activated cell sorting; BSA; DCs; GM-CSF; FCS; SEM
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2016.10.004

Dendritic cells (DCs) are increasingly being used for multiple applications and are useful tools for many immunotherapeutic strategies. The understanding of the possible impact of the DCs-generation methods on the biological capacities of these cells is therefore essential. Although the immunomagnetic separation is regarded as a fast and accurate method yielding cells with the high purity and efficiency, still little is known about its impact on the properties of the generated DCs. The aim of this study was to compare the morphology of the monocyte derived dendritic cells (MoDCs), generated from monocytes selected with anti-CD14 mAbs (positive separation) and treated with anti-CD3, -CD7, -CD16, -CD19, -CD56, -CD123, glycophorin A (negative separation), using laser scanning microscopy. We found that the type of the immunomagnetic separation method used strongly influences the shape and cell dimension of the MoDCs. We observed that the height of both immature and LPS-matured DCs generated from monocytes isolated by negative separation was significantly higher compared to the cells obtained by positive separation.
•The types of MACS separation influence the morphology of the MoDCs.•Significant differences are noted in dendritic cell height visualized by SEM.•Only at positive selection, the height of mature DC is lower than for immature DC.