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Mouse embryonic stem cell expansion in a microcarrier-based stirred culture system

2007; Elsevier BV; Volume: 132; Issue: 2 Linguagem: Inglês

10.1016/j.jbiotec.2007.05.031

1873-4863

Aline Marie Fernandes, Tiago G. Fernandes, Maria Margarida Diogo, Cláudia L. da Silva, Domingos Henrique, Joaquim M. S. Cabral,

Viral Infectious Diseases and Gene Expression in Insects

Embryonic stem (ES) cells have the ability to differentiate in vitro into a wide variety of cell types with potential applications for tissue regeneration. However, a large number of cells are required, thus strengthening the need to develop large-scale systems using chemically defined media for ES cell production and/or controlled differentiation. In the present studies, a stirred culture system (i.e. spinner flask) was used to scale-up mouse ES (mES) cell expansion in serum-containing (DMEM/FBS) or serum-free medium, both supplemented with leukemia inhibitory factor (LIF), using either Cytodex 3 or Cultispher S microcarriers. After 8 days, maximal cell densities achieved were (1.9 ± 0.1), (2.6 ± 0.7) and 3.5 × 106 cells/mL for Cytodex 3 in DMEM/FBS, Cultispher S in DMEM/FBS and Cultispher S in serum-free cultures, respectively, with fold increases of 38 ± 2, 50 ± 15 and 70. Both microcarriers were suitable to sustain mES cell expansion, though the macroporous Cultispher S seemed to be advantageous in providing a more protective environment against shear stress forces, which harmful effects are exacerbated in serum-free conditions. Importantly, mES cells expanded under stirred conditions using serum-free medium retained their pluripotency and the ability to commit to the neural lineage.