| Title | Cardiomyocyte Production in Mass Suspension Culture: Embryonic Stem Cells As a Source for Great Amounts of Functional Cardiomyocytes. | | Author(s) | Niebruegge S, Nehring A, Bär H, Schroeder M, Zweigerdt R, Lehmann J | | Institution | Institute of Cell Culture Technology, University of Bielefeld, Bielefeld, Germany., Institute of Biomaterials and Biomedical Engineering, Terrance Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada. | | Source | Tissue Eng Part A 2008 Jul 1. | | Abstract | Exploiting embryonic stem cell (ESC)-derived cardiomyocytes as a vital source for cell therapies and tissue engineering will depend on robust, large-scale production processes. Recently, we have reported stirring-controlled formation of embryoid bodies, enabling the generation of pure cardiomyocytes in 2-L scale. Expansion and differentiation of genetically engineered mouse ESCs was followed by antibiotic-based cardiomyocyte enrichment. Here we have investigated modification of various parameters aiming at process optimization in a 250-mL spinner flask system. Duration of the differentiation phase, timing of retinoic acid addition, and a slower medium exchange rate were found to be crucial to enhancing cardiomyocyte yield. Improved process conditions were consequently transferred to a 2-L controlled bioreactor. Employing a manual fill-and-draw medium change resulted in the formation of 0.86 x 10(9) cardiomyocytes in a single 2-L batch, thereby reproducing our previous findings. In contrast, an automated perfusion-based strategy enabled the production of 4.6 x 10(9) cardiomyocytes in a single run. This is significantly higher than previously reported and highlights the enormous process optimization potential in the scalable generation of ESC-derived cell lineages. | | Language | ENG | | Pub Type(s) | JOURNAL ARTICLE
| | PubMed ID | 18593268 |
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