Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells

• Type of Material: Article
• Authors: Maria C. Naskou, University of Georgia; Scarlett M. Sumner, University of Georgia; Anna Chocallo, University of Georgia; Hannah Kemelmakher, University of Georgia; Merrilee Thoresen, University of Georgia; Ian Copland, Emory University; Jacques Galipeau, Emory University; John F. Peroni, University of Georgia
• Language: English
• Date: 2018-03-22
• Publisher: BioMed Central
• Publication Version: Final Published Version
• Copyright Statement: © 2018 The Author(s).
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1186/s13287-018-0823-3
• Title of Journal or Parent Work: Stem Cell Research and Therapy
• ISSN: 1757-6512
• Volume: 9
• Issue: 1
• Start Page: 75
• End Page: 75
• Grant/Funding Information: This research was funded by the Morris Animal Foundation (grant number D17EQ-021).
• Keywords: Science & Technology; Life Sciences & Biomedicine; PROGENITOR CELLS; Cell Biology; ADIPOSE-TISSUE; CALF SERUM; Mesenchymal stem cells; RICH PLASMA; REGENERATIVE MEDICINE; STROMAL CELLS; Cell culture; Medicine, Research & Experimental; ANIMAL SERUM; Equine platelet lysate; THERAPY; IN-VITRO EXPANSION; Fetal bovine serum; Equine platelet apheresis; Research & Experimental Medicine; FETAL BOVINE SERUM
• Research Categories: Biology, Cell; Health Sciences, Oncology; Health Sciences, Medicine and Surgery

Abstract

Background: Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Methods: Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Results: Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed the release of TNF-α when exposed to LPS-stimulated monocytes similar to those cultured in FBS. Conclusion: ePL has the potential to be used for the expansion of MSCs before clinical application, avoiding the concerns associated with the use of FBS.