In vitro gene and chromosome characterization of expanded bone marrow mesenchymal stem cells for musculo-skeletal applications

OBJECTIVE : A number of studies have shown the role of expanded Bone Marrow- derived Mesenchymal Stem Cells in the repair and regeneration of musculo-skeletal tissues. The current European regulations define in vitro expanded cells for clinical purposes as substan - tially manipulated and include them in the class of Advanced-Therapy Medicinal Products to be manufactured in compliance with current Good Manufacturing Practice. Among the characteris - tics that such cells should display, genomic sta - bility has recently become a major safety con - cern. The aim of this study is to perform a chro - mosomal and genetic characterization of Bone Marrow-derived Mesenchymal Stem Cells ex - panded in compliance with Good Manufacturing Practice for a potential clinical use in or - thopaedics. MATERIALS AND METHODS: Mesenchymal Stem Cells, isolated from bone marrow, were ex - panded for six weeks in compliance with current Good Manufacturing Practice. DNA profiling analyses were applied to test cross-contamina - tion absence. Genomic stability was evaluated by means of karyotyping, sequencing of TP53, p21/CDKN1A and MDM2 genes and the expres - sion analysis of c-MYC and H-RAS oncogenes,