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Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood

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Abstract

Macrophages are among the most important cells of the immune system. Among other functions, they take part in almost all defense actions against foreign bodies and bacteria, being particularly important in infections, wound healing, and foreign body reactions. Considering their importance for the health of the human body, as well as their important role in several diseases, the in vitro studies based on these cells, are a crucial research field. Taking all mentioned into account, this study describes a simple isolation method of human macrophages (MFUM-HMP-001 and MFUM-HMP-002 cell lines) from peripheral blood. For this purpose, the morphology, the viability, and the phagocytotic activity of the isolated cells were tested. The Immunostaining of MFUM-HMP-001 and MFUM-HMP-002 cells confirmed the macrophage cell markers CD68, CD80, and CD163/M130. The phagocytotic activity was marked in both MFUM-HMP-001 and MFUM-HMP-002 cells, as was the phagocytosis of the pHrodo green Escherichia coli bioparticles conjugates, which was enhanced with the addition of lipopolysaccharide. The cells were stable and exhibited good growth. According to our results, both cell lines are useful for the development of novel macrophage cell-based in vitro models.

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Funding

The authors would like to acknowledge the financial support for this project received from the Slovenian Research Agency (Grant Nos: J3-1762, L4-1843, P3-0036 and I0-0029).

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Authors and Affiliations

  1. Faculty of Medicine, Institute of Biomedical Sciences, University of Maribor, Taborska ulica 8, 2000, Maribor, Slovenia

    Lidija Gradišnik, Marko Milojević, Tomaž Velnar & Uroš Maver

  2. AMEU-ECM Maribor, Slovenska 17, 2000, Maribor, Slovenia

    Lidija Gradišnik & Tomaž Velnar

  3. Department of Neurosurgery, University Medical Centre Ljubljana, Zaloska cesta 2, Ljubljana, Slovenia

    Tomaž Velnar

  4. Faculty of Medicine, Department of Pharmacology, University of Maribor, Taborska ulica 8, 2000, Maribor, Slovenia

    Uroš Maver

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  1. Lidija Gradišnik
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  2. Marko Milojević
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  3. Tomaž Velnar
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  4. Uroš Maver
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Contributions

LG., MM., TV., and UM. designed the study. LG. performed the experiments. LG., MM., TV., and UM. wrote the manuscript and prepared the figures. All authors reviewed the manuscript and gave the final approval for its submission.

Corresponding authors

Correspondence to Tomaž Velnar or Uroš Maver.

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The authors declare no conflict of interest.

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All experimental work was conducted only after suitable approvals by the Republic of Slovenia National Medical Ethics Committee were obtained (approval number 0120-674/2015-4) and obtained written consents of the included persons.

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Gradišnik, L., Milojević, M., Velnar, T. et al. Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood. Mol Biol Rep 47, 6929–6940 (2020). https://doi.org/10.1007/s11033-020-05751-6

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  • DOI: https://doi.org/10.1007/s11033-020-05751-6

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