The Effect of Autologous Platelet Lysate on the Expansion and Adhesion Characteristics of Equine Bone Marrow-Derived Mesenchymal Stem Cells
Abstract
Mesenchymal stem cells (MSCs) are undifferentiated cells that are being investigated as a therapy for equine musculoskeletal disease. Fetal bovine serum (FBS) is commonly used as a serum supplement when culturing MSCs. However, there is a risk of xenogenic reactions when MSCs that have incorporated bovine proteins are injected into a patient. Researchers have begun searching for autologous alternatives to FBS, including platelet lysate (PL). Platelets and their growth factors might allow for cell proliferation without the risk of xenogenic reactions. The impact of autologous platelet lysate supplementation on the expansion and tissue culture plastic adhesion of equine bone marrow-derived MSCs was assessed. MSC proliferation in 10% serum supplemented media was compared using PL and FBS as sources of serum. A 10% FBS medium containing heparin was also tested as a control for heparin in the PL supplement. MSC proliferation was assessed using cell counts and CellTrace™ Violet proliferation assays. Preliminary data showed greater overall population numbers in the PL and FBS with heparin supplemented mediums compared to FBS alone. Population doubling times were lower for PL and FBS heparin supplemented mediums compared to FBS alone early in cell culture time. However, at later cell culture times, population doubling times were higher for PL-supplemented cultures. The amount of cells lost during routine media removal and washes of the cell monolayer was assessed through a DNA assay. Preliminary data showed greater cell loss in PL and FBS heparin supplemented cultures as compared to FBS alone, especially during times when PL population doubling times had increased. Flow cytometry was used to assay for the presence of cell surface markers. All groups were positive for cell surface markers CD-90 and negative for CD-45 and MHCII. The high cell losses in PL-supplemented cultures during times when PL population doubling times increased indicates that cell losses may be obscuring the population growth rates for these MSCs. If this is the case, then solving the adhesion problem in PL-supplemented MSC cultures may allow these cultures to grow to higher cell numbers at a higher rate.
Subject
mesenchymal stem cellsplatelet lysate
fetal bovine serum
autologous serum supplementation
cell proliferation
Citation
Donald, Brittany Anne Nicole (2016). The Effect of Autologous Platelet Lysate on the Expansion and Adhesion Characteristics of Equine Bone Marrow-Derived Mesenchymal Stem Cells. Undergraduate Research Scholars Program. Available electronically from https : / /hdl .handle .net /1969 .1 /196652.