期刊
JOURNAL OF BIOTECHNOLOGY
卷 146, 期 4, 页码 194-197出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2010.02.015
关键词
Mesenchymal stem cells; Expansion; Microcarriers; Spinner flask; Low-serum content medium; Cell metabolism
资金
- Associacao Portuguesa contra a Leucemia
- MIT-Portugal
- Spanish Ministry of Education and Science [2007-0109]
- Fundacao para a Ciencia e Tecnologia, Portugal [SFRH/BD/38719/2007, SFRH/BD/38720/2007]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/38719/2007, SFRH/BD/38720/2007] Funding Source: FCT
Bioreactor systems have been developed as alternatives to standard culture flasks clue 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. (C) 2010 Elsevier B.V. All rights reserved.
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