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Scale‐up of mouse embryonic stem cell expansion in stirred bioreactors
Author(s) -
FernandesPlatzgummer Ana,
Diogo Maria M.,
Baptista Ricardo P.,
Silva Cláudia Lobato da,
Cabral Joaquim M.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.658
Subject(s) - microcarrier , laboratory flask , bioreactor , embryonic stem cell , homeobox protein nanog , stem cell , chemistry , cell culture , chemically defined medium , microbiology and biotechnology , biology , cell , andrology , biochemistry , in vitro , induced pluripotent stem cell , botany , medicine , genetics , gene
The aim of this study was to develop a robust, quality controlled and reproducible large‐scale culture system using serum‐free (SF) medium to obtain vast numbers of embryonic stem (ES) cells as a starting source for potential applications in tissue regeneration, as well as for drug screening studies. Mouse ES (mES) cells were firstly cultured on microcarriers in spinner flasks to investigate the effect of different parameters such as the agitation rate and the feeding regimen. Cells were successfully expanded at agitation rates up to 60 rpm using the SF medium and no significant differences in terms of growth kinetics or metabolic profiles were found between the two feeding regimens evaluated: 50% medium renewal every 24 h or 25% every 12 h. Overall, cells reached maximum concentrations of (4.2 ± 0.4) and (5.6 ± 0.8) ×10 6 cells/mL at Day 8 for cells fed once or twice per day; which corresponds to an increase in total cell number of 85 ± 7 and 108 ± 16, respectively. To have a more precise control over culture conditions and to yield a higher number of cells, the scale‐up of the spinner flask culture system was successfully accomplished by using a fully controlled stirred tank bioreactor. In this case, the concentration of mES cells cultured on microcarriers increased 85 ± 15‐fold over 11 days. Importantly, mES cells expanded under stirred conditions, in both spinner flask and fully controlled stirred tank bioreactor, using SF medium, retained the expression of pluripotency markers such as Oct‐4, Nanog, and SSEA‐1 and their differentiation potential into cells of the three embryonic germ layers. © 2011 American Institute of Chemical Engineers Biotechnol. Prog.,, 2011