title: Controlling expansion and cardiomyogenic differentiation of human pluripotent stem cells in scalable suspension culture.
abstract: To harness the potential of human pluripotent stem cells (hPSCs), an abundant supply of their progenies is required. Here, hPSC expansion as matrix-independent aggregates in suspension culture was combined with cardiomyogenic differentiation using chemical Wnt pathway modulators. A multiwell screen was scaled up to stirred Erlenmeyer flasks and subsequently to tank bioreactors, applying controlled feeding strategies (batch and cyclic perfusion). Cardiomyogenesis was sensitive to the GSK3 inhibitor CHIR99021 concentration, whereas the aggregate size was no prevailing factor across culture platforms. However, in bioreactors, the pattern of aggregate formation in the expansion phase dominated subsequent differentiation. Global profiling revealed a culture-dependent expression of BMP agonists/antagonists, suggesting their decisive role in cell-fate determination. Furthermore, metallothionein was discovered as a potentially stress-related marker in hPSCs. In 100 ml bioreactors, the production of 40 million predominantly ventricular-like cardiomyocytes (up to 85% purity) was enabled that were directly applicable to bioartificial cardiac tissue formation.
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Henning
authors.initials: H
authors.lastname: Kempf
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; Member of the Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Olmer
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Kropp
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Rückert
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Monica
authors.initials: M
authors.lastname: Jara-Avaca
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Robles-Diaz
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Annika
authors.initials: A
authors.lastname: Franke
authors.affiliation: Murdoch Childrens Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, VIC 3052, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Elliott
authors.affiliation: Institute for Neurophysiology, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Wojciechowski
authors.affiliation: Institute for Neurophysiology, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Fischer
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Angelica
authors.initials: A
authors.lastname: Roa Lara
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: George
authors.initials: G
authors.lastname: Kensah
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Ina
authors.initials: I
authors.lastname: Gruh
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Haverich
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; Member of the Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation, and Vascular Surgery (HTTG), Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany; REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Straβe 1, 30625 Hannover, Germany. Electronic address: zweigerdt.robert@mh-hannover.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Zweigerdt
mesh_terms: D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003470:Culture Media; D017077:Culture Media, Conditioned; D006801:Humans; D008564:Membrane Potentials; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D003470:Culture Media; D017077:Culture Media, Conditioned