title: Efficient suspension bioreactor expansion of murine embryonic stem cells on microcarriers in serum-free medium.
abstract: Large numbers of cells will be required for successful embryonic stem cell (ESC)-based cellular therapies or drug discovery, thus raising the need to develop scaled-up bioprocesses for production of ESCs and their derived progeny. Traditionally, ESCs have been propagated in adherent cultures in static flasks on fibroblasts layers in serum-containing medium. Direct translation of two-dimensional flatbed cultures to large-scale production of the quantities of cells required for therapy simply by increasing the number of dishes or flasks is not practical or economical. Here, we describe successful scaled-up production of ESCs on microcarriers in a stirred culture system in a serum-free medium. Cells expanded on CultiSpher S, Cytodex 3, and Collagen microcarriers showed superior cell-fold expansions of 439, 193, and 68, respectively, without excessive agglomeration, compared with 27 in static culture. In addition, the ESCs maintained their pluripotency after long-term culture (28 days) in serum-free medium. This is the first time mESCs have been cultured on microcarriers without prior exposure to serum and/or fibroblasts, while also eliminating the excessive agglomeration plaguing earlier studies. These protocols provide an economical, practical, serum-free means for expanding ESCs in a stirred suspension bioprocess.
authors.affiliation: Pharmaceutical Production Research Facility, Schulich School of Engineering, University of Calgary, Calgary, AB, Canada T2N 1N4.
authors.forename: Roz
authors.initials: R
authors.lastname: Alfred
authors.forename: Jaret
authors.initials: J
authors.lastname: Radford
authors.forename: Jessica
authors.initials: J
authors.lastname: Fan
authors.forename: Kathryn
authors.initials: K
authors.lastname: Boon
authors.forename: Roman
authors.initials: R
authors.lastname: Krawetz
authors.forename: Derrick
authors.initials: D
authors.lastname: Rancourt
authors.forename: Michael S
authors.initials: MS
authors.lastname: Kallos
mesh_terms: D000818:Animals; D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D049109:Cell Proliferation; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology*; D051379:Mice; D013535:Suspensions
chemical_list: D016895:Culture Media, Serum-Free; D013535:Suspensions