Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches: Further insights in the search for a fetal calf serum substitute

• Published in: Journal of cellular physiology Vol. 211; no. 1; pp. 121 - 130
• Main Authors: Bernardo, M.E., Avanzini, M.A., Perotti, C., Cometa, A.M., Moretta, A., Lenta, E., Del Fante, C., Novara, F., de Silvestri, A., Amendola, G., Zuffardi, O., Maccario, R., Locatelli, F.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.04.2007
• Subjects: Adolescent; CD4 Antigens - immunology; Cell Count; Cell Differentiation; Cell Proliferation; Cell- and Tissue-Based Therapy; Cells, Cultured; Colony-Forming Units Assay; Cytokines - secretion; Cytotoxicity, Immunologic; Fibroblasts - cytology; Humans; Interleukin-2 Receptor alpha Subunit - immunology; Karyotyping; Killer Cells, Natural - cytology; Killer Cells, Natural - immunology; Kinetics; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - immunology; Multipotent Stem Cells - cytology; Phenotype; Serum - metabolism; Stromal Cells - cytology; T-Lymphocytes - cytology; T-Lymphocytes - immunology
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.20911

There is great interest in mesenchymal stromal cells (MSCs) for cell‐therapy and tissue engineering approaches. MSCs are currently expanded in vitro in the presence of fetal calf serum (FCS); however, FCS raises concerns when used in clinical grade preparations. The aim of this study was to evaluate whether MSCs expanded in medium supplemented with platelet‐lysate (PL), already shown to promote MSC growth, are endowed with biological properties appropriate for cell‐therapy approaches. We confirm previously published data showing that MSCs expanded in either FCS or PL display comparable morphology, phenotype, and differentiation capacity, while PL‐MSCs were superior in terms of clonogenic efficiency and proliferative capacity. We further extended these data by investigating the immune‐regulatory effect of MSCs on the alloantigen‐specific immune response in mixed lymphocyte culture (MLC). We found that MSCs‐PL are comparable to MSCs‐FCS in their capacity to: (i) decrease alloantigen‐induced cytotoxic activity; (ii) favor differentiation of CD4+ T‐cell subsets expressing a Treg phenotype; (iii) increase early secretion of IL‐10 in MLC supernatant, as well as induce a striking augmentation of IL‐6 production. As compared with MSCs‐PL, MSCs‐FCS were more efficient in suppressing alloantigen‐induced lymphocyte subset proliferation and reducing early IFNγ‐secretion. Resistance to spontaneous transformation into tumor cells of expanded MSCs was demonstrated by molecular karyotyping and maintenance of normal morphology/phenotype after prolonged in vitro culture. Our data support the immunological functional plasticity of MSCs and suggest that MSCs‐PL can be used as an alternative to MSCs‐FCS, although these latter cells might be more suitable for preventing/treating alloreactivity‐related immune complications. J. Cell. Physiol. 211: 121–130, 2007. © 2006 Wiley‐Liss, Inc.