Jiye Gao1, Maaike R Scheenstra2, Albert van Dijk2, Edwin J A Veldhuizen2, Henk P Haagsman3. 1. Division of Molecular Host Defence, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands; Rongchang Campus, Southwest University, Chongqing, China. 2. Division of Molecular Host Defence, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. 3. Division of Molecular Host Defence, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. Electronic address: H.P.Haagsman@uu.nl.
Abstract
BACKGROUND: Macrophages play an important role in the innate immune system as part of the mononuclear phagocyte system (MPS). They have a pro-inflammatory signature (M1-polarized macrophages) or anti-inflammatory signature (M2-polarized macrophages) based on expression of surface receptors and secretion of cytokines. However, very little is known about the culture of macrophages from pigs and more specific about the M1 and M2 polarization in vitro. METHODS: Porcine monocytes or mononuclear bone marrow cells were used to culture M1- and M2-polarized macrophages in the presence of GM-CSF and M-CSF, respectively. Surface receptor expression was measured with flow cytometry and ELISA was used to quantify cytokine secretion in response to LPS and PAM3CSK4 stimulation. Human monocyte-derived macrophages were used as control. RESULTS: Porcine M1- and M2-polarized macrophages were cultured best using porcine GM-CSF and murine M-CSF, respectively. Cultures from bone marrow cells resulted in a higher yield M1- and M2-polarized macrophages which were better comparable to human monocyte-derived macrophages than cultures from porcine monocytes. Porcine M1-polarized macrophages displayed the characteristic fried egg shape morphology, lower CD163 expression and low IL-10 production. Porcine M2-polarized macrophages contained the spindle-like morphology, higher CD163 expression and high IL-10 production. CONCLUSION: Porcine M1- and M2-polarized macrophages can be most efficiently cultured from mononuclear bone marrow cells using porcine GM-CSF and murine M-CSF. The new culture method facilitates more refined studies of porcine macrophages in vitro, important for both porcine and human health since pigs are increasingly used as model for translational research.
BACKGROUND: Macrophages play an important role in the innate immune system as part of the mononuclear phagocyte system (MPS). They have a pro-inflammatory signature (M1-polarized macrophages) or anti-inflammatory signature (M2-polarized macrophages) based on expression of surface receptors and secretion of cytokines. However, very little is known about the culture of macrophages from pigs and more specific about the M1 and M2 polarization in vitro. METHODS: Porcine monocytes or mononuclear bone marrow cells were used to culture M1- and M2-polarized macrophages in the presence of GM-CSF and M-CSF, respectively. Surface receptor expression was measured with flow cytometry and ELISA was used to quantify cytokine secretion in response to LPS and PAM3CSK4 stimulation. Human monocyte-derived macrophages were used as control. RESULTS: Porcine M1- and M2-polarized macrophages were cultured best using porcine GM-CSF and murineM-CSF, respectively. Cultures from bone marrow cells resulted in a higher yield M1- and M2-polarized macrophages which were better comparable to human monocyte-derived macrophages than cultures from porcine monocytes. Porcine M1-polarized macrophages displayed the characteristic fried egg shape morphology, lower CD163 expression and low IL-10 production. Porcine M2-polarized macrophages contained the spindle-like morphology, higher CD163 expression and high IL-10 production. CONCLUSION: Porcine M1- and M2-polarized macrophages can be most efficiently cultured from mononuclear bone marrow cells using porcine GM-CSF and murineM-CSF. The new culture method facilitates more refined studies of porcine macrophages in vitro, important for both porcine and human health since pigs are increasingly used as model for translational research.
Authors: Vijaykumar S Meli; Ryan P Donahue; Jarrett M Link; Jerry C Hu; Kyriacos A Athanasiou; Wendy F Liu Journal: Biomater Sci Date: 2021-11-23 Impact factor: 6.843
Authors: Federica Marinaro; María Gómez-Serrano; Inmaculada Jorge; Juan Carlos Silla-Castro; Jesús Vázquez; Francisco Miguel Sánchez-Margallo; Rebeca Blázquez; Esther López; Verónica Álvarez; Javier G Casado Journal: Front Bioeng Biotechnol Date: 2019-12-20
Authors: Elvira Ruslanovna Akhmetzyanova; Anna Viktorovna Timofeeva; Davran Khudaishukurovich Sabirov; Alexander Alexandrovich Kostennikov; Alexander Alexandrovich Rogozhin; Victoria James; S S Arkhipova; Albert Anatolevich Rizvanov; Yana Olegovna Mukhamedshina Journal: Front Mol Neurosci Date: 2022-02-24 Impact factor: 5.639
Authors: Ryan P Donahue; Jarrett M Link; Vijaykumar S Meli; Jerry C Hu; Wendy F Liu; Kyriacos A Athanasiou Journal: Cartilage Date: 2022 Jan-Mar Impact factor: 3.117
Authors: Jesús Pérez-Ortega; Roel M Van Harten; Ria Van Boxtel; Michel Plisnier; Marc Louckx; Dominique Ingels; Henk P Haagsman; Jan Tommassen Journal: Virulence Date: 2021-12 Impact factor: 5.882