title: Bone-forming capacity of mesenchymal stromal cells when cultured in the presence of human platelet lysate as substitute for fetal bovine serum.
abstract: In tissue engineering, strategies are being developed to repair large bone defects by combining biomaterials and bone marrow-derived multipotent mesenchymal stromal cells (MSCs). For expansion of MSCs under good manufacturing practice conditions, human platelet lysate (PL) can serve as substitute for fetal bovine serum (FBS) in culture media. We compared the in vivo bone-forming capacity of passage 3 MSCs cultured with either PL or FBS for nine different human donors. We also tested the growth kinetics, antigen expression profile, and the multilineage differentiation capacity in vitro of these MSCs. The in vivo bone-forming capacity was determined by seeding culture-expanded MSCs onto biphasic calcium phosphate scaffolds. Hybrid constructs were implanted subcutaneously in nude mice, retrieved after 6 weeks, and analyzed using histomorphometry. PL-supplemented cultures resulted in significantly larger colonies, shorter culture time period, and higher population doublings between P1 and P3 compared to FBS-containing cultures. No differences were observed in antigen expression profiles or differentiation capacities into the osteoblastic, chondrogenic, and adipogenic lineages, qualitatively. In vivo bone formation with PL-supplemented cultures of MSCs was demonstrated in 9/9 donors versus 6/9 for FBS-supplemented cultures. These results warrant the use of PL for ex vivo expansion of human MSCs for bone tissue engineering applications.
authors.affiliation: Department of Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Henk-Jan
authors.initials: HJ
authors.lastname: Prins
authors.forename: Henk
authors.initials: H
authors.lastname: Rozemuller
authors.forename: Simone
authors.initials: S
authors.lastname: Vonk-Griffioen
authors.forename: Vivienne G M
authors.initials: VG
authors.lastname: Verweij
authors.forename: Wouter J A
authors.initials: WJ
authors.lastname: Dhert
authors.forename: Ineke C M
authors.initials: IC
authors.lastname: Slaper-Cortenbach
authors.forename: Anton C M
authors.initials: AC
authors.lastname: Martens
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology; D001792:Blood Platelets / Q000166:cytology*; D001802:Blood Substitutes / Q000494:pharmacology; D002417:Cattle; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002457:Cell Extracts / Q000494:pharmacology*; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D006801:Humans; D016130:Immunophenotyping; D007700:Kinetics; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D010012:Osteogenesis / Q000187:drug effects*; D044967:Serum / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D000941:Antigens; D001802:Blood Substitutes; D002457:Cell Extracts