title: Expansion of embryonic stem cells in suspension and fibrous bed bioreactors.
abstract: Applications of embryonic stem (ES) cells in cellular transplantation and tissue engineering require scalable processes for mass production of these cells with controlled qualities. The main objective of this work was to evaluate two cell culture processes for long-term expansion of murine embryonic stem (mES) cells. With serial passaging, suspension cultures in spinner flasks were able to expand mES cells as aggregates for 12.5-fold in each passage of 4 days. However, extending the culturing time to 6 days in each passage caused significant loss in cell viability and induced differentiation as indicated by the reduced expression levels of SSEA-1 and Oct-4. Long-term expansion of mES cells in a fibrous bed bioreactor (FBB) was also studied for 30 days in 2 passages, 15 days in each passage. With periodically refreshing the culture medium, a high expansion fold of 60-77 was achieved in each passage. Flow cytometry and RT-PCR were used to analyze key pluripotency and differentiation markers. The results showed that the expanded cells in both suspension and FBB cultures remained in a highly pluripotent state, which was also confirmed with the embryoid body (EB) forming efficiency test. It is concluded that both the suspension and FBB cultures are suitable to support long-term expansion of undifferentiated mES cells. However, the FBB culture can sustain cell growth for a longer period without frequent passaging, requires less media and labor, and is thus more economical to use for mass production of ES cells.
authors.affiliation: William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.affiliation: Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, 2525 Pottsdamer Street, Tallahassee, FL 32310, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA. Electronic address: yang.15@osu.edu.
authors.forename: Shang-Tian
authors.initials: ST
authors.lastname: Yang
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D058732:Embryoid Bodies / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D019344:Lactic Acid / Q000378:metabolism; D051379:Mice
chemical_list: D019344:Lactic Acid; D005947:Glucose
keywords: Embryonic stem cells; Expansion; Fibrous bed bioreactor; Suspension culture; Three-dimensional culture