title: Equine platelet lysate as an alternative to fetal bovine serum in equine mesenchymal stromal cell culture - too much of a good thing?
abstract: BACKGROUND
  Multipotent mesenchymal stromal cells (MSC) are often culture-expanded in vitro. Presently, expansion medium (EM) for MSC is supplemented with fetal bovine serum (FBS). However, increasing cost, variable composition and potential risks associated with bovine antigens call for alternatives. Platelet lysate (PL) has shown promise as an alternative supplement.
  
  
  OBJECTIVE
  To determine how equine umbilical cord blood (CB) MSC proliferate in EM enriched with PL or FBS at various concentrations.
  
  
  METHODS
  Randomised dose escalation study.
  
  
  METHODS
  Platelet concentrate was generated from 5 equine whole blood samples through a double centrifugation method and standardised to 1 × 10(12) platelets/l prior to a freeze/thaw cycle to produce PL. Pooled PL or pooled FBS was added to EM at concentrations of 5% to 60%. Proliferation of 4 equine CB-MSC cultures was determined after 4 days using a resazurin semiquantitative assay.
  
  
  RESULTS
  Cord blood-MSC proliferated with a dose-dependent response with no significant difference found between PL and FBS up to a 30% concentration. Beyond 30%, proliferation fell in the PL-cultured cells, while continued dose-dependent proliferation was noted in the FBS-cultured cells. Despite reduced cell numbers in high PL concentrations, live/dead staining revealed that adherent cells remained viable.
  
  
  CONCLUSIONS
  Expansion medium enriched with PL can support short-term equine CB-MSC proliferation at conventional culture concentrations. Based on the unexpected suppression of CB-MSC at higher PL concentrations, an in vivo dose study is indicated to investigate if combinational therapies of CB-MSC and platelet-rich plasma are associated with synergistic or antagonistic effect on CB-MSC function.
authors.affiliation: Department of Biomedical Sciences, University of Guelph, Ontario, Canada.
authors.forename: K A
authors.initials: KA
authors.lastname: Russell
authors.affiliation: Department of Biomedical Sciences, University of Guelph, Ontario, Canada.|Department of Clinical Studies, Orthopaedic Research Lab, Aarhus University, Copenhagen, Denmark.
authors.forename: T G
authors.initials: TG
authors.lastname: Koch
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D002417:Cattle; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D003470:Culture Media; D006736:Horses; D059630:Mesenchymal Stem Cells / Q000502:physiology*
chemical_list: D003470:Culture Media
keywords: blood plasma; cell culture technique; cord blood; horse; stem cell