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Efficient suspension bioreactor expansion of murine embryonic stem cells on microcarriers in serum‐free medium
Author(s) -
Alfred Roz,
Radford Jaret,
Fan Jessica,
Boon Kathryn,
Krawetz Roman,
Rancourt Derrick,
Kallos Michael S.
Publication year - 2011
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.591
Subject(s) - microcarrier , laboratory flask , embryonic stem cell , bioprocess , microbiology and biotechnology , bioreactor , cell culture , suspension culture , chemically defined medium , chemistry , biology , immunology , biochemistry , in vitro , genetics , botany , paleontology , gene
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. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011