title: Derivation, expansion and differentiation of induced pluripotent stem cells in continuous suspension cultures.
abstract: We describe derivation of induced pluripotent stem cells (iPSCs) from terminally differentiated mouse cells in serum- and feeder-free stirred suspension cultures. Temporal analysis of global gene expression revealed high correlations between cells reprogrammed in suspension and cells reprogrammed in adhesion-dependent conditions. Suspension culture-reprogrammed iPSCs (SiPSCs) could be differentiated into all three germ layers in vitro and contributed to chimeric embryos in vivo. SiPSC generation allowed for efficient selection of reprogramming factor-expressing cells based on their differential survival and proliferation in suspension culture. Seamless integration of SiPSC reprogramming and directed differentiation enabled scalable production of beating cardiac cells in a continuous single cell- and small aggregate-based process. This method is an important step toward the development of robust PSC generation, expansion and differentiation technology.
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: David A
authors.initials: DA
authors.lastname: Fluri
authors.forename: Peter D
authors.initials: PD
authors.lastname: Tonge
authors.forename: Hannah
authors.initials: H
authors.lastname: Song
authors.forename: Ricardo P
authors.initials: RP
authors.lastname: Baptista
authors.forename: Nika
authors.initials: N
authors.lastname: Shakiba
authors.forename: Shreya
authors.initials: S
authors.lastname: Shukla
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Clarke
authors.forename: Andras
authors.initials: A
authors.lastname: Nagy
authors.forename: Peter W
authors.initials: PW
authors.lastname: Zandstra
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D065150:Cellular Reprogramming / Q000502:physiology; D002678:Chimera / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*