title: Growth and functional harvesting of human mesenchymal stromal cells cultured on a microcarrier-based system.
abstract: Human mesenchymal stromal cells (hMSCs) cells are attractive for applications in tissue engineering and cell therapy. Because of the low availability of hMSCs in tissues and the high doses of hMSCs necessary for infusion, scalable and cost-effective technologies for in vitro cell expansion are needed to produce MSCs while maintaining their functional, immunophenotypic and cytogenetic characteristics. Microcarrier-based culture systems are a good alternative to traditional systems for hMSC expansion. The aim of the present study was to develop a scalable bioprocess for the expansion of human bone marrow mesenchymal stromal cells (hBM-MSCs) on microcarriers to optimize growth and functional harvesting. In general, the results obtained demonstrated the feasibility of expanding hBM-MSCs using microcarrier technology. The maximum cell concentration (n = 5) was ~4.82 ± 1.18 × 10(5) cell mL(-1) at day 7, representing a 3.9-fold increase relative to the amount of inoculated cells. At the end of culture, 87.2% of the cells could be harvested (viability = 95%). Cell metabolism analysis revealed that there was no depletion of important nutrients such as glucose and glutamine during culture, and neither lactate nor ammonia byproducts were formed at inhibitory concentrations. The cells that were recovered after the expansion retained their immunophenotypic and functional characteristics. These results represent an important step toward the implementation of a GMP-compliant large-scale production system for hMSCs for cellular therapy.
authors.affiliation: Hemotherapy Center of Ribeirão Preto, School of Medicine of Ribeirão Preto, University of São Paulo, Tenente Catão Roxo Street, 2501, CEP 14051-140, Ribeirão Preto-SP, Brazil.
authors.forename: Sâmia R
authors.initials: SR
authors.lastname: Caruso
authors.forename: Maristela D
authors.initials: MD
authors.lastname: Orellana
authors.forename: Amanda
authors.initials: A
authors.lastname: Mizukami
authors.forename: Taisa R
authors.initials: TR
authors.lastname: Fernandes
authors.forename: Aparecida M
authors.initials: AM
authors.lastname: Fontes
authors.forename: Claudio A T
authors.initials: CA
authors.lastname: Suazo
authors.forename: Viviane C
authors.initials: VC
authors.lastname: Oliveira
authors.forename: Dimas T
authors.initials: DT
authors.lastname: Covas
authors.forename: Kamilla
authors.initials: K
authors.lastname: Swiech
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D064987:Cell- and Tissue-Based Therapy; D003470:Culture Media / Q000737:chemistry*; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D003470:Culture Media; D019344:Lactic Acid
keywords: cell expansion; functional harvest; mesenchymal stromal cells; microcarrier culture; spinner flasks