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The identification of markers of macrophage differentiation in PMA-stimulated THP-1 Cells and monocyte-derived macrophages

Daigneault, M., Preston, J.A., Marriott, H.M., Whyte, M.K.B. and Dockrell, D.H. (2010) The identification of markers of macrophage differentiation in PMA-stimulated THP-1 Cells and monocyte-derived macrophages. Plos One, 5 (1). Art no.e8668. ISSN 1932-6203

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Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response. The phenotype of mature tissue macrophages represents the composite of environmental and differentiation-dependent imprinting. Phorbol-12-myristate-13-acetate (PMA) and 1,25-dihydroxyvitamin D3 (VD3) are stimuli commonly used to induce macrophage differentiation in monocytic cell lines but the extent of differentiation in comparison to primary tissue macrophages is unclear. We have compared the phenotype of the promonocytic THP-1 cell line after various protocols of differentiation utilising VD3 and PMA in comparison to primary human monocytes or monocyte-derived macrophages (MDM). Both stimuli induced changes in cell morphology indicative of differentiation but neither showed differentiation comparable to MDM. In contrast, PMA treatment followed by 5 days resting in culture without PMA (PMAr) increased cytoplasmic to nuclear ratio, increased mitochondrial and lysosomal numbers and altered differentiation-dependent cell surface markers in a pattern similar to MDM. Moreover, PMAr cells showed relative resistance to apoptotic stimuli and maintained levels of the differentiation-dependent anti-apoptotic protein Mcl-1 similar to MDM. PMAr cells retained a high phagocytic capacity for latex beads, and expressed a cytokine profile that resembled MDM in response to TLR ligands, in particular with marked TLR2 responses. Moreover, both MDM and PMAr retained marked plasticity to stimulus-directed polarization. These findings suggest a modified PMA differentiation protocol can enhance macrophage differentiation of THP-1 cells and identify increased numbers of mitochondria and lysosomes, resistance to apoptosis and the potency of TLR2 responses as important discriminators of the level of macrophage differentiation for transformed cells.

Item Type: Article
Copyright, Publisher and Additional Information: © 2010 Daigneault et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords: Tumor-Necrosis-Factor; Human Alveolar Macrophages; Mononuclear Phagocytes; Blood Monocytes; Nitric-Oxide; Factor-Alpha; Streptococcus-Pneumoniae; Bronchoalveolar Lavage; Vitro Differentiation; Dendritic Cells
Institution: The University of Sheffield
Academic Units: The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Medicine (Sheffield)
Depositing User: Miss Anthea Tucker
Date Deposited: 23 Feb 2010 11:46
Last Modified: 04 Jun 2014 17:24
Published Version: http://dx.doi.org/10.1371/journal.pone.0008668
Status: Published
Publisher: Public Library Science
Identification Number: 10.1371/journal.pone.0008668
URI: http://eprints.whiterose.ac.uk/id/eprint/10432

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