期刊
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
卷 18, 期 1, 页码 173-184出版社
KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
DOI: 10.1007/s12257-012-0512-5
关键词
ex vivo expansion; bone marrow mesenchymal stem cells; suspension culture; bioreactor
资金
- National High Technology Research and Development Program of China (863 Program) [2012AA021201]
- Fundamental Research Funds for the Central Universities (ECUST)
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 x 10(6) 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.
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