title: Maximizing the ex vivo expansion of human mesenchymal stem cells using a microcarrier-based stirred culture system.
abstract: Bioreactor systems have been developed as alternatives to standard culture flasks due to their homogeneous nature, easiness of monitoring and increased cell production. Here we investigated the in vitro expansion of bone marrow (BM) mesenchymal stem cells (MSC) in spinner flasks, using gelatin microcarriers (Cultispher S) to support cell adhesion and proliferation. MSC expansion was performed using a low-serum containing medium (2% of fetal bovine serum, FBS). A strategy was defined for the maximization of cell expansion: microcarriers were pre-coated with FBS in order to increase cell seeding efficiency and an adequate feeding regime was established (25% medium exchange everyday). The maximum cell density, 4.2 x 10(5)cells/mL, was obtained at day 8, corresponding to a fold increase in total cell number of 8.4+/-0.8. Expanded MSC retained their differentiation potential into adipogenic and osteogenic lineages, as well as their clonogenic ability. Harvested cells expressed >90% of CD73, CD90 and CD105 markers. These results demonstrated that a microcarrier-based stirred culture system is adequate for human MSC expansion, using a low-serum containing medium, allowing the generation of significant cell numbers for potential applications in regenerative medicine.
authors.affiliation: IBB - Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.
authors.forename: Gemma
authors.initials: G
authors.lastname: Eibes
authors.forename: Francisco
authors.initials: F
authors.lastname: dos Santos
authors.forename: Pedro Z
authors.initials: PZ
authors.lastname: Andrade
authors.forename: Joana S
authors.initials: JS
authors.lastname: Boura
authors.forename: Manuel M A
authors.initials: MM
authors.lastname: Abecasis
authors.forename: Cláudia Lobato
authors.initials: CL
authors.lastname: da Silva
authors.forename: Joaquim M S
authors.initials: JM
authors.lastname: Cabral
mesh_terms: D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D055442:Metabolome
chemical_list: D003470:Culture Media