期刊
BIOTECHNOLOGY PROGRESS
卷 27, 期 3, 页码 811-823出版社
WILEY
DOI: 10.1002/btpr.591
关键词
embryonic stem cells; suspension bioreactor; microcarriers; serum-free cultivation
资金
- Province of Alberta (Queen Elizabeth II Scholarship)
- Canadian Institutes of Health Research (CIHR)
- NSERC
- NIH [5R21AR53738-2]
- Canadian Stem Cell Network Centre of Excellence
Large numbers of cells will be required for successful embryonic stem cell (ESC)-based cellular therapies or drug discovery, thus raising the need to develop scaled-up bioprocesses for production of ESCs and their derived progeny. Traditionally, ESCs have been propagated in adherent cultures in static flasks on fibroblasts layers in serum-containing medium. Direct translation of two-dimensional flatbed cultures to large-scale production of the quantities of cells required for therapy simply by increasing the number of dishes or flasks is not practical or economical. Here, we describe successful scaled-up production of ESCs on microcarriers in a stirred culture system in a serum-free medium. Cells expanded on CultiSpher S, Cytodex 3, and Collagen microcarriers showed superior cell-fold expansions of 439, 193, and 68, respectively, without excessive agglomeration, compared with 27 in static culture. In addition, the ESCs maintained their pluripotency after long-term culture (28 days) in serum-free medium. This is the first time mESCs have been cultured on microcarriers without prior exposure to serum and/or fibroblasts, while also eliminating the excessive agglomeration plaguing earlier studies. These protocols provide an economical, practical, serum-free means for expanding ESCs in a stirred suspension bioprocess. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 811-823, 2011
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