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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.
J Cell Physiol. 2007 Apr; 211(1):121-30.JC

Abstract

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 IFNgamma-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.

Authors+Show Affiliations

Oncoematologia Pediatrica, IRCCS Policlinico San Matteo, Università di Pavia, Pavia, Italy. mebernardo@gmail.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17187344

Citation

Bernardo, M E., et al. "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." Journal of Cellular Physiology, vol. 211, no. 1, 2007, pp. 121-30.
Bernardo ME, Avanzini MA, Perotti C, et al. 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. J Cell Physiol. 2007;211(1):121-30.
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. (2007). 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. Journal of Cellular Physiology, 211(1), 121-30.
Bernardo ME, et al. 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. J Cell Physiol. 2007;211(1):121-30. PubMed PMID: 17187344.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Bernardo,M E, AU - Avanzini,M A, AU - Perotti,C, AU - Cometa,A M, AU - Moretta,A, AU - Lenta,E, AU - Del Fante,C, AU - Novara,F, AU - de Silvestri,A, AU - Amendola,G, AU - Zuffardi,O, AU - Maccario,R, AU - Locatelli,F, PY - 2006/12/26/pubmed PY - 2007/4/5/medline PY - 2006/12/26/entrez SP - 121 EP - 30 JF - Journal of cellular physiology JO - J. Cell. Physiol. VL - 211 IS - 1 N2 - 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 IFNgamma-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. SN - 0021-9541 UR - https://www.unboundmedicine.com/medline/citation/17187344/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_ L2 - https://doi.org/10.1002/jcp.20911 DB - PRIME DP - Unbound Medicine ER -