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Mouse embryonic stem cell expansion in a microcarrier-based stirred culture system
A M Fernandes1, T G Fernandes, M M Diogo
1Institute for Biotechnology and Bioengineering (IBB), Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.
Journal of Biotechnology
|July 24, 2007
Summary
Large-scale expansion of mouse embryonic stem cells (mESCs) is crucial for regenerative medicine. Stirred cultures with serum-free media and Cultispher S microcarriers efficiently produced pluripotent mESCs, retaining their differentiation potential.
Area of Science:
- Stem Cell Biology
- Biotechnology
- Tissue Engineering
Background:
- Embryonic stem (ES) cells offer potential for tissue regeneration but require large-scale production systems.
- Chemically defined media are needed for scalable ES cell expansion and controlled differentiation.
Purpose of the Study:
- To develop and evaluate a stirred culture system for large-scale mouse embryonic stem cell (mES) expansion.
- To compare the efficacy of different microcarriers and media conditions (serum-containing vs. serum-free) for mES cell production.
Main Methods:
- Mouse ES cells were cultured in spinner flasks using Cytodex 3 or Cultispher S microcarriers.
- Cultures were maintained in either standard DMEM/FBS medium or a serum-free medium, both supplemented with leukemia inhibitory factor (LIF).
- Cell density and pluripotency were assessed after 8 days.
Main Results:
- Maximal cell densities of 3.5x10^6 cells/mL were achieved using Cultispher S in serum-free medium, with a 70-fold increase.
- Cultispher S microcarriers provided a protective environment against shear stress, especially in serum-free conditions.
- mES cells expanded in serum-free stirred cultures retained pluripotency and neural lineage commitment ability.
Conclusions:
- Stirred culture systems, particularly with Cultispher S microcarriers and serum-free media, enable efficient large-scale expansion of mouse ES cells.
- This scalable method maintains mES cell pluripotency and differentiation capacity, supporting regenerative medicine applications.
