Ex vivo expansion of bone marrow mesenchymal stem cells using microcarrier beads in a stirred bioreactor
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Bone marrow mesenchymal stem cells (bmMSCs) have recently gained attention as a useful resource in the fields of regenerative medicine and tissue engineering. However, the number of bmMSCs obtained from available donors is very low. Here we developed a culture strategy for in vitro expansion of bmMSCs in a 1.5 L stirred bioreactor with microcarrier beads. First, the microcarriers (Cytodex 3) were equilibrated in culture medium containing 3% fetal bovine serum (FBS) for at least 30 min prior to cell addition. After inoculation, the FBS concentration of the medium was maintained at 3% (v/v) in the first 24 h and thereafter maintained at 1% (v/v) and a developed feeding regimen was applied over 5 days. The maximum cell density of 2.6 × 106 cells/mL was achieved at day 5, corresponding to a 10.4 ± 0.8 fold increases in total cell number. Among the harvested cells, 98.95% expressed CD29 and 84.48% expressed CD90, suggesting that the majority of expanded bmMSCs still retained their differentiation potential. Therefore, the developed microcarrier-based stirred bioreactor culture system is an effective method to generate significant numbers of bmMSCs for potential applications and research studies.
Keywordsex vivo expansion bone marrow mesenchymal stem cells suspension culture bioreactor
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- 18.Dos Santos, F., P. Z. Andrade, J. S. Boura, M. Abecasis, C. L. da Silva, and J. M. S. Cabral (2010) Ex vivo expansion of human mesenchymal stem cells: A more effective cell proliferation kinetics and metabolism under hypoxia. J. Cellular Physiol. 223: 27–35.Google Scholar