title: Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.
abstract: Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates.
authors.affiliation: Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil.
authors.forename: Amanda
authors.initials: A
authors.lastname: Mizukami
authors.affiliation: Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Fernandes-Platzgummer
authors.affiliation: Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Joana G
authors.initials: JG
authors.lastname: Carmelo
authors.affiliation: Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil. kamilla@fcfrp.usp.br.|Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil. kamilla@fcfrp.usp.br.
authors.forename: Kamilla
authors.initials: K
authors.lastname: Swiech
authors.affiliation: Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil.
authors.forename: Dimas T
authors.initials: DT
authors.lastname: Covas
authors.affiliation: Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Joaquim M S
authors.initials: JM
authors.lastname: Cabral
authors.affiliation: Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal . claudia_lobato@tecnico.ulisboa.pt.
authors.forename: Cláudia L
authors.initials: CL
authors.lastname: da Silva
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors; D002452:Cell Count; D002454:Cell Differentiation; D049109:Cell Proliferation; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology; D014470:Umbilical Cord / Q000166:cytology* / Q000276:immunology
keywords: Mesenchymal stem/stromal cells; Microcarriers; Serum-/xenogeneic-free culture medium; Stirred-tank bioreactor; Umbilical cord matrix