title: Suspension culture of human pluripotent stem cells in controlled, stirred bioreactors.
abstract: Therapeutic and industrial applications of pluripotent stem cells and their derivatives require large cell quantities generated in defined conditions. To this end, we have translated single cell-inoculated suspension cultures of human pluripotent stem cells (hPSCs; including human induced pluripotent stem cells [hiPS] and human embryonic stem cells [hESC]) to stirred tank bioreactors. These systems that are widely used in biopharmaceutical industry allow straightforward scale up and detailed online monitoring of key process parameters. To ensure minimum medium consumption, but in parallel functional integration of all probes mandatory for process monitoring, that is, for pO₂ and pH, experiments were performed in 100 mL culture volume in a "mini reactor platform" consisting of four independently controlled vessels. By establishing defined parameters for tightly controlled cell inoculation and aggregate formation up to 2×10⁸ hiPSCs/100 mL were generated in a single process run in 7 days. Expression of pluripotency markers and ability of cells to differentiate into derivates of all three germ layers in vitro was maintained, underlining practical utility of this new process. The presented data provide key steps toward scalable mass expansion of human iPS and ES cells thereby enabling translation of stem cell research to (pre)clinical application in relevant large animal models and valuable in vitro assays for drug development and validation as well.
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs-LEBAO, Department of Cardiac, Thoracic, Transplantation, and Vascular Surgery, Hannover, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Olmer
authors.forename: Andreas
authors.initials: A
authors.lastname: Lange
authors.forename: Sebastian
authors.initials: S
authors.lastname: Selzer
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kasper
authors.forename: Axel
authors.initials: A
authors.lastname: Haverich
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
authors.forename: Robert
authors.initials: R
authors.lastname: Zweigerdt
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D019149:Bioreactors; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D048429:Cell Size; D002478:Cells, Cultured; D005855:Germ Layers / Q000166:cytology; D006801:Humans; D007700:Kinetics; D039904:Pluripotent Stem Cells / Q000166:cytology*; D015203:Reproducibility of Results; D013535:Suspensions
chemical_list: D015415:Biomarkers; D013535:Suspensions