title: Large-scale expansion of pluripotent human embryonic stem cells in stirred-suspension bioreactors.
abstract: Since the derivation of human embryonic stem (hES) cells, their translation to clinical therapies has been met with several challenges, including the need for large-scale expansion and controlled differentiation processes. Suspension bioreactors are an effective alternative to static culture flasks as they enable the generation of clinically relevant cell numbers with greater efficacy in a controlled culture system. We, along with other groups, have developed bioreactor protocols for the expansion of pluripotent murine ES cells. Here we present a novel bioreactor protocol that yields a 25-fold expansion of hES cells over 6 days. Using immunofluorescence, flow cytometry, and teratoma formation assays, we demonstrated that these bioreactor cultures retained high levels of pluripotency and a normal karyotype. Importantly, the use of bioreactors enables the expansion of hES cells in the absence of feeder layers or matrices, which will facilitate the adaptation of good manufacturing process (GMP) standards to the development of hES cell therapies.
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Roman
authors.initials: R
authors.lastname: Krawetz
authors.forename: Jaymi T
authors.initials: JT
authors.lastname: Taiani
authors.forename: Shiying
authors.initials: S
authors.lastname: Liu
authors.forename: Guoliang
authors.initials: G
authors.lastname: Meng
authors.forename: Xiangyun
authors.initials: X
authors.lastname: Li
authors.forename: Michael S
authors.initials: MS
authors.lastname: Kallos
authors.forename: Derrick E
authors.initials: DE
authors.lastname: Rancourt
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020123:Sirolimus / Q000494:pharmacology; D013535:Suspensions
chemical_list: D047428:Protein Kinase Inhibitors; D013535:Suspensions; D020123:Sirolimus