Research Paper

Biotechnology and Bioprocess Engineering

, Volume 18, Issue 1, pp 173-184

Ex vivo expansion of bone marrow mesenchymal stem cells using microcarrier beads in a stirred bioreactor

  • Lang ZhouAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
  • , Jiantao KongAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
  • , Yingping ZhuangAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
  • , Ju ChuAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
  • , Siliang ZhangAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
  • , Meijin GuoAffiliated withState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology Email author 

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Abstract

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.

Keywords

ex vivo expansion bone marrow mesenchymal stem cells suspension culture bioreactor