Impact of individual platelet lysates on isolation and growth of human mesenchymal stromal cells |
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| Authors: | Patrick Horn Gudrun Bokermann Dominik Cholewa Simone Bork Thomas Walenda Carmen Koch Wolf Drescher Gabriele Hutschenreuther Martin Zenke Anthony D. Ho Wolfgang Wagner |
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| Affiliation: | 1. Centro di Ricerca Tettamanti, Clinica Pediatrica, Università di Milano-Bicocca, Monza, Italy;2. Istituto di ricovero e cura a carattere scientifico-Istituto di Ricerche Farmacologiche “Mario Negri”, Milano, Italy;3. Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy;4. Humanitas Clinical and Research Center, Rozzano, Italy;5. Clinica Pediatrica, Università di Milano-Bicocca, Fondazione Monza e Brianza per il Bambino e la sua Mamma/Ospedale S. Gerardo, Monza, Italy;1. Biotherapy Division, Macopharma, Mouvaux, France;2. PAnTher, Institut National de la Recherche Agronomique (INRA), Ecole Nationale Vétérinaire, Agro-alimentaire et de l''Alimentation Nantes-Atlantique (Oniris), Université Bretagne Loire, Nantes, France;3. Université de Nantes, Université Bretagne Loire, Nantes, France;4. Medical Department, Macopharma, Tourcoing, France |
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| Abstract: | Background aimsCulture medium for mesenchymal stromal cells (MSC) is frequently supplemented with fetal calf serum (FCS). FCS can induce xenogeneic immune reactions, transmit bovine pathogens and has a high lot-to-lot variability that hampers reproducibility of results. Several studies have demonstrated that pooled human platelet lysate (HPL) provides an attractive alternative for FCS. However, little is known about the variation between different platelet lysates.MethodsWe compared activities of individual HPL on initial fibroblastoid colony-forming units (CFU-F), proliferation, in vitro differentiation and long-term culture. These data were correlated with chemokine profiles of HPL.ResultsIsolation of MSC with either HPL or FCS resulted in similar CFU-F frequency, colony morphology, immunophenotype and adipogenic differentiation potential. Osteogenic differentiation was even more pronounced in HPL than FCS. There were significant differences in MSC proliferation with different HPL, but it was always higher in comparison with FCS. Cell growth correlated with the concentration of platelet-derived growth factor (PDGF) and there was a moderate association with platelet counts. All HPL facilitated expansion for more than 20 population doublings.ConclusionsTaken together, reliable long-term expansion was possible with all HPL, although there was some variation in platelet lysates of individual units. Therefore the use of donor recipient-matched or autologous HPL is feasible for therapeutic MSC products. |
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