The authors declare no competing financial interests.
Communication to the Editor
High density continuous production of murine pluripotent cells in an acoustic perfused bioreactor at different oxygen concentrations†
Article first published online: 18 SEP 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 2, pages 648–655, February 2013
How to Cite
Baptista, R. P., Fluri, D. A. and Zandstra, P. W. (2013), High density continuous production of murine pluripotent cells in an acoustic perfused bioreactor at different oxygen concentrations. Biotechnol. Bioeng., 110: 648–655. doi: 10.1002/bit.24717
- Issue published online: 20 DEC 2012
- Article first published online: 18 SEP 2012
- Accepted manuscript online: 4 SEP 2012 11:06AM EST
- Manuscript Accepted: 24 AUG 2012
- Manuscript Revised: 16 AUG 2012
- Manuscript Received: 4 JUN 2012
- Natural Sciences and Engineering Research Council of Canada
- Canadian Institute of Health Research. Grant Number: MOP-57885
- McLean Award
- acoustic filter;
- murine pluripotent cells;
Strategies for the production of pluripotent stem cells (PSCs) rely on serially dissociated adherent or aggregate-based culture, consequently limiting robust scale-up of cell production, on-line control and optimization of culture conditions. We recently developed a method that enables continuous (non-serially dissociated) suspension culture-mediated reprogramming to pluripotency. Herein, we use this method to demonstrate the scalable production of PSCs and early derivatives using acoustic filter technology to enable continuous oxygen-controlled perfusion culture. Cell densities of greater than 1 × 107 cells/mL were achieved after 7 days of expansion at a specific growth rate (µ) of 0.61 ± 0.1 day−1 with a perfusion rate (D) of 5.0 day−1. A twofold increase in maximum cell density (to greater than 2.5 × 107 cells/mL) was achieved when the medium dissolved oxygen was reduced (5% DO). Cell densities and viabilities >80% were maintained for extended production periods during which maintenance of pluripotency was confirmed by stable expression of pluripotency factors (SSEA-1 and Nanog), as well as the capacity to differentiate into all three germ layers. This work establishes a versatile biotechnological platform for the production of pluripotent cells and derivatives in an integrated, scalable and intensified stirred suspension culture. Biotechnol. Bioeng. 2013; 110: 648–655. © 2012 Wiley Periodicals, Inc.