In vitro biosafety profile evaluation of multipotent mesenchymal stem cells derived from the bone marrow of sarcoma patients

Journal of Translational Medicine

Serena Duchi³  Davide Donati¹  Daniela Lisini⁶  Luca Sangiorgi⁵  Martina Pandolfi⁵  Monia Zuntini⁵  Orsetta Zuffardi⁴  Giulia Arrigo⁴  Francesca Novara⁴  Rita Maccario⁷  Maria Antonietta Avanzini⁷  Melissa Mantelli⁷  Chiara Bellotti³  Enrico Lucarelli² 

[1] Osteoarticular Regeneration Laboratory, 3rd Clinic of Orthopaedics and Traumatology, Rizzoli Orthopaedic Institute, Bologna, Italy;Osteoarticolar Regeneration Laboratory, Rizzoli Orthopaedic Institute, Bologna, Italy;Department of Biomedical and Neuromotor Sciences (DIBINEM), Alma Mater Studiorum University of Bologna, Bologna, Italy;Department of Molecular Medicine, University of Pavia, Pavia, Italy;Department of Medical Genetics and Skeletal Rare Diseases, Rizzoli Orthopaedic Institute, Bologna, Italy;Cell Therapy Production Unit (UPTC), IRCCS Neurologic Institute C. Besta Foundation, Milano, Italy;Immunology and Transplantation Laboratory/Cell Factory/ Pediatric Oncohematology, Fondazione IRCCS Policlinico S. Matteo, Pavia, Italy

关键词: Tumorigenic transformation;    Tissue regeneration;    Osteosarcoma (OS);    Mesenchymal stem cells (MSCs);    Ewing sarcoma (EWS);   

Others  :  815078
DOI  :  10.1186/1479-5876-12-95

 received in 2014-02-21, accepted in 2014-03-31,  发布年份 2014

【 摘 要 】

Background

In osteosarcoma (OS) and most Ewing sarcoma (EWS) patients, the primary tumor originates in the bone. Although tumor resection surgery is commonly used to treat these diseases, it frequently leaves massive bone defects that are particularly difficult to be treated. Due to the therapeutic potential of mesenchymal stem cells (MSCs), OS and EWS patients could benefit from an autologous MSCs-based bone reconstruction. However, safety concerns regarding the in vitro expansion of bone marrow-derived MSCs have been raised. To investigate the possible oncogenic potential of MSCs from OS or EWS patients (MSC-SAR) after expansion, this study focused on a biosafety assessment of MSC-SAR obtained after short- and long-term cultivation compared with MSCs from healthy donors (MSC-CTRL).

Methods

We initially characterized the morphology, immunophenotype, and differentiation multipotency of isolated MSC-SAR. MSC-SAR and MSC-CTRL were subsequently expanded under identical culture conditions. Cells at the early (P3/P4) and late (P10) passages were collected for the in vitro analyses including: sequencing of genes frequently mutated in OS and EWS, evaluation of telomerase activity, assessment of the gene expression profile and activity of major cancer pathways, cytogenetic analysis on synchronous MSCs, and molecular karyotyping using a comparative genomic hybridization (CGH) array.

Results

MSC-SAR displayed comparable morphology, immunophenotype, proliferation rate, differentiation potential, and telomerase activity to MSC-CTRL. Both cell types displayed signs of senescence in the late stages of culture with no relevant changes in cancer gene expression. However, cytogenetic analysis detected chromosomal anomalies in the early and late stages of MSC-SAR and MSC-CTRL after culture.

Conclusions

Our results demonstrated that the in vitro expansion of MSCs does not influence or favor malignant transformation since MSC-SAR were not more prone than MSC-CTRL to deleterious changes during culture. However, the presence of chromosomal aberrations supports rigorous phenotypic, functional and genetic evaluation of the biosafety of MSCs, which is important for clinical applications.

【 授权许可】

   
2014 Lucarelli et al.; licensee BioMed Central Ltd.

【 图 表 】

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