Innate immune responses of equine monocytes cultured in equine platelet lysate.
Abstract: Platelet lysate (PL) has been extensively used for the laboratory expansion of human mesenchymal stem cells (MSC) in order to avoid fetal bovine serum (FBS) which has been associated with immune-mediated host reactions and transmission of bovine-origin microbial contaminants. Before suggesting the routine use of PL for MSC culture, we wanted to further investigate whether PL alone might trigger inflammatory responses when exposed to reactive white blood cells such as monocytes. Our objectives were to evaluate the inflammatory profile of equine monocytes cultured with equine PL (ePL) and to determine if ePL can modulate the expression of inflammatory cytokines in lipopolysaccharide (LPS)-stimulated monocytes. In a first experiment, equine monocytes were isolated and incubated with donor horse serum (DHS), FBS, six individual donors ePL or pooled ePL from all horses. In a second experiment, monocytes were stimulated with E. coli LPS in the presence of 1, 5 or 10% DHS and/or pooled ePL. After 6h of incubation, cell culture supernatants were assayed via ELISA for production of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and Interleukin 1β (IL-1β) as well as for the anti-inflammatory Interleukin 10 (IL-10). Equine monocytes incubated with pooled ePL produced significantly less TNF-α and significantly more IL-10 than monocytes incubated in FBS. A statistically significant difference was not identified for the production of IL-1β. The second experiment showed that pooled ePL added to LPS-stimulated equine monocytes resulted in a significant reduction in TNF-α and IL-1β production. IL-10 production was not significantly upregulated by the addition of ePL to LPS-stimulated monocytes. Finally, the addition of ePL to LPS-stimulated monocytes in the presence of various concentrations of DHS resulted to statistically significant decrease of TNF-α and IL-1β compared to the control groups. This is the first study to demonstrate that ePL suppresses the release of pro-inflammatory cytokines from stimulated equine monocytes. These results encourage further exploration of PL as a homologous media substitute for FBS but also opens the possibility of investigating its use as means to suppress cell-mediated inflammation.
Published by Elsevier B.V.
Publication Date: 2017-11-16 PubMed ID: 29249319DOI: 10.1016/j.vetimm.2017.11.005Google Scholar: Lookup
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Summary
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The study investigates the impact of equine platelet lysate (ePL) on the inflammatory responses of equine monocytes and explores its potential use as a substitute medium for fetal bovine serum (FBS) in mesenchymal stem cells (MSCs) culture. Results indicate that ePL suppresses the release of certain pro-inflammatory cytokines and triggers an increased production of anti-inflammatory cytokines, suggesting its potential utility in suppressing cell-mediated inflammation.
Background and Objectives
- The research aimed to explore the use of platelet lysate (PL) as an alternative to fetal bovine serum (FBS) for expanding human mesenchymal stem cells (MSC) in laboratories. This is because FBS has been linked to immune-related host reactions and possible transmission of bovine-origin microbes.
- It specifically examined whether ePL could provoke inflammatory responses when exposed to reactive equine monocytes – a type of white blood cell. Researchers also studied if ePL can alter the expression of inflammatory cytokines when subjected to lipopolysaccharide (LPS)-stimulated monocytes.
Research Methods
- The team conducted two experiments. First, they isolated equine monocytes and incubated them with donor horse serum (DHS), FBS, or ePL from six individual donors, or a pooled ePL from all horses.
- In the second experiment, monocytes were stimulated with E. coli LPS in the presence of different concentrations of DHS and/or pooled ePL. Cell culture supernatants were then evaluated via ELISA for the production of pro-inflammatory cytokines (TNF-α and IL-1β) and the anti-inflammatory cytokine IL-10.
Findings
- Monocytes incubated with pooled ePL had significantly lower levels of the pro-inflammatory cytokine TNF-α, but they produced significantly more anti-inflammatory cytokine IL-10 compared to monocytes incubated in FBS. There was no significant statistical difference in the generation of IL-1β.
- In the second experiment, the addition of ePL to LPS-stimulated monocytes significantly reduced the production of both TNF-α and IL-1β. There was no significant change in IL-10 production with the addition of ePL.
- The study indicated that ePL could suppress the release of pro-inflammatory cytokines from stimulated equine monocytes.
Implications
- The findings suggest further exploration of using PL as a substitute media for FBS in MSC cultures is warranted, given its potential for reducing inflammatory responses.
- The study also opens possibilities for investigating the use PL to suppress cell-mediated inflammation, which could be useful in inflammation-related conditions or diseases.
Cite This Article
APA
Naskou MC, Norton NA, Copland IB, Galipeau J, Peroni JF.
(2017).
Innate immune responses of equine monocytes cultured in equine platelet lysate.
Vet Immunol Immunopathol, 195, 65-71.
https://doi.org/10.1016/j.vetimm.2017.11.005 Publication
Researcher Affiliations
- Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA 30602, United States.
- Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA 30602, United States.
- Emory Personalized Immunotherapy Center [EPIC], Emory University School of Medicine, 100 Woodruff Circle Atlanta, GA 30322, United States.
- Department of Medicine and Carbone Comprehensive Cancer Center, University of Wisconsin, 600 Highland Ave, Madison, WI 53792, United States.
- Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA 30602, United States. Electronic address: jperoni@uga.edu.
MeSH Terms
- Animals
- Blood Platelets / metabolism
- Cells, Cultured
- Culture Media
- Female
- Horses / immunology
- Immunity, Innate / immunology
- Immunity, Innate / physiology
- Male
- Monocytes / immunology
- Monocytes / physiology
Citations
This article has been cited 12 times.- Bowlby CM, Purmessur D, Durgam SS. Equine peripheral blood CD14(+) monocyte-derived macrophage in-vitro characteristics after GM-CSF pretreatment and LPS+IFN-γ or IL-4+IL-10 differentiation. Vet Immunol Immunopathol 2023 Jan;255:110534.
- Even KM, Gaesser AM, Ciamillo SA, Linardi RL, Ortved KF. Comparing the immunomodulatory properties of equine BM-MSCs culture expanded in autologous platelet lysate, pooled platelet lysate, equine serum and fetal bovine serum supplemented culture media. Front Vet Sci 2022;9:958724.
- Rampin A, Skoufos I, Raghunath M, Tzora A, Diakakis N, Prassinos N, Zeugolis DI. Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture. Cells 2022 May 5;11(9).
- Gordon J, Álvarez-Narváez S, Peroni JF. Antimicrobial Effects of Equine Platelet Lysate. Front Vet Sci 2021;8:703414.
- Gilbertie JM, Schaer TP, Schubert AG, Jacob ME, Menegatti S, Ashton Lavoie R, Schnabel LV. Platelet-rich plasma lysate displays antibiofilm properties and restores antimicrobial activity against synovial fluid biofilms in vitro. J Orthop Res 2020 Jun;38(6):1365-1374.
- Gilbertie JM, Long JM, Schubert AG, Berglund AK, Schaer TP, Schnabel LV. Pooled Platelet-Rich Plasma Lysate Therapy Increases Synoviocyte Proliferation and Hyaluronic Acid Production While Protecting Chondrocytes From Synoviocyte-Derived Inflammatory Mediators. Front Vet Sci 2018;5:150.
- Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF. Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther 2018 Mar 22;9(1):75.
- Berghaus LJ, Venner M, Helbig H, Hildebrandt D, Hart K. The potential value of cytokine, cortisol and vitamin D profiles in foals from birth to weaning for respiratory disease prediction on a farm endemic for Rhodococcus equi pneumonia. Equine Vet J 2026 Mar;58(2):359-371.
- Hildebrandt D, Venner M, Hart KA, Berghaus L. Plasma C-reactive protein and interleukin-6 concentrations in foals during health and respiratory disease. Equine Vet J 2026 Mar;58(2):372-379.
- Mollabashi M, Klopfer A, Lunardon T, Darzenta N, Davis E, Murray M, Sumner SM, Naskou MC. Plasma and complement proteins are essential for the antimicrobial activity of canine platelet lysate. Front Vet Sci 2025;12:1605649.
- Lunardon T, Sumner SM, Mollabashi M, Darzenta N, Davis E, Naskou MC. Growth factor and cytokine characterization of canine platelet lysate with variable leukocyte concentration, plasma content, and heat-sensitive proteins. Front Vet Sci 2024;11:1408080.
- Morales-Vázquez MM, Meza-Serrano E, Lara-Pereyra I, Acuña-González RJ, Alonso-Morales R, Hayen-Valles S, Boeta AM, Zarco L, Lozano-Cuenca J, López-Canales JS, Flores-Herrera H. Equine Placentitis in Mares Induces the Secretion of Pro-Inflammatory Cytokine eIL-1β and the Active Extracellular Matrix Metalloproteinase (MMP)-9. Vet Sci 2023 Aug 22;10(9).
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