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KMID : 1094720130180010173
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 1 p.173 ~ p.184
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Ex vivo expansion of bone marrow mesenchymal stem cells using microcarrier beads in a stirred bioreactor
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Zhou Lang
Kong Jiantao
Zhuang Yingping
Chu Ju
Zhang Siliang
Guo Meijin
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Abstract
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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.
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KEYWORD
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ex vivo expansion, bone marrow mesenchymal stem cells, suspension culture, bioreactor
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