Large-scale expansion of mouse embryonic stem cells on microcarriers.
A large-scale stirred culture system for the expansion of mouse embryonic stem cells (mESCs) in spinner flasks under serum-free conditions was established using macroporous microcarriers for cell attachment and growth. This type of microcarrier was chosen as it potentially offers more protection to cells against shear stress in the absence of serum compared to microporous ones. In addition, methods to characterize ESCs after large-scale expansion were established. The pluripotency of expanded mESCs was evaluated based on both flow cytometry and alkaline phosphatase staining. Envisaging the application of ESCs as a potential source of neural progenitors, the neural commitment potential of cells after expansion in the spinner flask was also determined by culturing cells in serum-free adherent monolayer conditions.
Centre for Biological and Chemical Engineering, Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Lisboa, Portugal.
Ana
A
Fernandes-Platzgummer
Maria Margarida
MM
Diogo
Cláudia Lobato
CL
da Silva
Joaquim M S
JM
Cabral
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D019070:Cell Lineage; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D051379:Mice; D008863:Microspheres; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling
D015415:Biomarkers; D016895:Culture Media, Serum-Free; D000469:Alkaline Phosphatase