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A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate
Authors:Barbara Dozza  Giuliana Gobbi  Enrico Lucarelli  Michela Pierini  Claudia Di Bella  Tommaso Frisoni  Pier Luigi Tazzari  Francesca Ricci  Prisco Mirandola  Cecilia Carubbi  Sandro Giannini  Davide Donati  Marco Vitale
Institution:1. Bone Regeneration Laboratory, 2nd Clinic of Orthopaedics and Traumatology, Rizzoli Orthopaedic Institute, , Bologna, Italy;2. Department of Anatomy, Pharmacology & Forensic Medicine, Human Anatomy Section, University of Parma, , Parma, Italy;3. Service of Transfusion Medicine, S.Orsola‐Malpighi Hospital, , Bologna, Italy
Abstract:Mesenchymal stem cells (MSCs) and platelet‐rich plasma (PRP) are currently used alone or in combination for therapeutic applications especially for bone repair. We tested whether MSCs can be isolated from bone marrow (BM) aspirate using a commercially available kit commonly used to obtain PRP from peripheral blood (PB). Results revealed that mononuclear cells and platelets from both PB and BM could be efficiently isolated by obtaining a mononuclear and platelet rich fraction (PB‐MPRF and BM‐MPRF, respectively). Starting with comparable volumes, the number of platelets increased 1.5‐fold in BM‐MPRF compared to PB‐MPRF. The number of clonogenic cells in BM‐MPRF samples was significantly higher than whole BM samples as revealed by CFU‐F assay (54.92 ± 8.55 CFU‐F/1.5 x 105 nucleated cells and 32.50 ± 12.43 CFU‐F/1.5 x 105 nucleated cells, respectively). Cells isolated from BM‐MPRF after in vitro expansion fulfilled the definition of MSCs by phenotypic criteria, and differentiated along osteogenic, adipogenic and chondrogenic lineages following induction. Results showed that the kit isolated MSCs and platelets from BM aspirate. Isolated MSCs were further expanded in a laboratory and BM‐MPRF was used clinically following BM withdrawal for rapid intra‐operative cell therapy for the treatment of bone defects. Copyright © 2012 John Wiley & Sons, Ltd.
Keywords:bone marrow aspirate  mesenchymal stem cells  osteocompetent cells  platelets  platelet‐rich plasma
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