Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood
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ORIGINAL ARTICLE
Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood Lidija Gradišnik1,2 · Marko Milojević1 · Tomaž Velnar1,2,3 · Uroš Maver1,4 Received: 19 February 2020 / Accepted: 28 August 2020 © Springer Nature B.V. 2020
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. Keywords Macrophages · Cell isolation protocol · Human peripheral blood · Cell characterization · Phagocytotic activity
Introduction As one of the body’s main defense systems, the immune response is the first line of protection against the pathogenic organisms and foreign bodies [1]. It acts in response to both tissue trauma and biomaterial implantation [2]. Any * Tomaž Velnar [email protected] * Uroš Maver [email protected] Lidija Gradišnik [email protected] 1
Faculty of Medicine, Institute of Biomedical Sciences, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia
2
AMEU-ECM Maribor, Slovenska 17, 2000 Maribor, Slovenia
3
Department of Neurosurgery, University Medical Centre Ljubljana, Zaloska cesta 2, Ljubljana, Slovenia
4
Faculty of Medicine, Department of Pharmacology, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia
infection, wounding or biomaterial insertion into the body tissue causes its damage and consequent response, which are also the first steps during the healing process. The latter begins at the moment of injury and involves both resident and migratory cell populations, as well as the active participation of the extracellular matrix and the action of soluble mediators. The humoral and cellular inflammatory responses aim to form an immune barrier against invading microorganisms or materials [3]. Among the most notable cells with an innate immune function are the macrophages. As effector cells, they take part in virtually all defense
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