title: Scalable expansion of human pluripotent stem cells in suspension culture.
abstract: Routine commercial and clinical applications of human pluripotent stem cells (hPSCs) and their progenies will require increasing cell quantities that cannot be provided by conventional adherent culture technologies. Here we describe a straightforward culture protocol for the expansion of undifferentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) in suspension culture. This culture technique was successfully tested on two hiPSC clones, three hESC lines and on a nonhuman primate ESC line. It is based on a defined medium and single-cell inoculation, but it does not require culture preadaptation, use of microcarriers or any other matrices. Over a time course of 4-7 d, hPSCs can be expanded up to sixfold. Preparation of a high-density culture and its subsequent translation to scalable stirred suspension in Erlenmeyer flasks and stirred spinner flasks are also feasible. Importantly, hPSCs maintain pluripotency and karyotype stability for more than ten passages.
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department of Cardiac, Thoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany. zweigerdt.robert@mh-hannover.de
authors.forename: Robert
authors.initials: R
authors.lastname: Zweigerdt
authors.forename: Ruth
authors.initials: R
authors.lastname: Olmer
authors.forename: Harmeet
authors.initials: H
authors.lastname: Singh
authors.forename: Axel
authors.initials: A
authors.lastname: Haverich
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*