title: Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components.
abstract: OBJECTIVE
  The clinical use of human mesenchymal stromal cells (MSC) requires ex vivo expansion in media containing supplements such as fetal bovine serum or, alternatively, human platelet lysate (PL).
  
  
  METHODS
  Platelet concentrates were frozen, quarantine stored, thawed and sterile filtered to obtain PL. PL content and its effect on fibroblast-colony-forming unit (CFU-F) formation, MSC proliferation and large-scale expansion were studied.
  
  
  RESULTS
  PL contained high levels of basic fibroblast growth factor (bFGF), soluble CD40L (sCD40L), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), platelet-derived growth factor AA (PDGF-AA), platelet-derived growth factor AB/BB (PDGF-AB/BB), chemokine (C-C) ligand 5 (CCL5; RANTES) transforming growth factor-β1 (TGF-β1) and chemokine (C-X-C) ligand 1/2/3 (GRO), with low batch-to-batch variability, and most were stable for up to 14 days. Inhibition of PDGF-BB and bFGF decreased MSC proliferation by about 20% and 50%, respectively. The strongest inhibition (about 75%) was observed with a combination of anti-bFGF + anti-PDGF-BB and anti-bFGF + anti-TGF-β1 + anti-PDGF-BB. Interestingly, various combinations of recombinant PDGF-BB, bFGF and TGF-β1 were not sufficient to promote cell proliferation. PL from whole blood-derived pooled platelet concentrates and apheresis platelet concentrates did not differ significantly in their growth-promoting activity on MSC.
  
  
  CONCLUSIONS
  PL enhances MSC proliferation and can be regarded as a safe tool for MSC expansion for clinical purposes. \in particular, PDGF-BB and bFGF are essential components for the growth-promoting effect of PL, but are not sufficient for MSC proliferation.
authors.affiliation: Institut für Transfusionsmedizin, Universität Ulm und Institut für Klinische Transfusionsmedizin und Immungenetik Ulm, DRK-Blutspendedienst Baden-Württemberg-Hessen, Germany.
authors.forename: Natalie
authors.initials: N
authors.lastname: Fekete
authors.forename: Mélanie
authors.initials: M
authors.lastname: Gadelorge
authors.forename: Daniel
authors.initials: D
authors.lastname: Fürst
authors.forename: Caroline
authors.initials: C
authors.lastname: Maurer
authors.forename: Julia
authors.initials: J
authors.lastname: Dausend
authors.forename: Sandrine
authors.initials: S
authors.lastname: Fleury-Cappellesso
authors.forename: Volker
authors.initials: V
authors.lastname: Mailänder
authors.forename: Ramin
authors.initials: R
authors.lastname: Lotfi
authors.forename: Anita
authors.initials: A
authors.lastname: Ignatius
authors.forename: Luc
authors.initials: L
authors.lastname: Sensebé
authors.forename: Philippe
authors.initials: P
authors.lastname: Bourin
authors.forename: Hubert
authors.initials: H
authors.lastname: Schrezenmeier
authors.forename: Markus Thomas
authors.initials: MT
authors.lastname: Rojewski
mesh_terms: D000818:Animals; D000077214:Becaplermin; D001781:Blood Component Removal; D001792:Blood Platelets / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D018925:Chemokines / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D020574:Proto-Oncogene Proteins c-sis / Q000008:administration & dosage*; D011994:Recombinant Proteins / Q000008:administration & dosage*
chemical_list: D018925:Chemokines; D016207:Cytokines; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D000077214:Becaplermin