title: Cardiomyocyte production in mass suspension culture: embryonic stem cells as a source for great amounts of functional cardiomyocytes.
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.
authors.affiliation: Institute of Cell Culture Technology, University of Bielefeld, Bielefeld, Germany. sylvia.niebruegge@gmail.com
authors.forename: Sylvia
authors.initials: S
authors.lastname: Niebruegge
authors.forename: Andrea
authors.initials: A
authors.lastname: Nehring
authors.forename: Harald
authors.initials: H
authors.lastname: Bär
authors.forename: Magnus
authors.initials: M
authors.lastname: Schroeder
authors.forename: Robert
authors.initials: R
authors.lastname: Zweigerdt
authors.forename: Juergen
authors.initials: J
authors.lastname: Lehmann
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000166:cytology*; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D032383:Myocytes, Cardiac / Q000166:cytology*