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Long-term microcarrier suspension cultures of human embryonic stem cells

2009; Elsevier BV; Volume: 2; Issue: 3 Linguagem: Inglês

10.1016/j.scr.2009.02.005

1876-7753

Steve Oh, Allen K. Chen, Yanglin Mok, Xiaoli Chen, U-Ming Lim, Angela Chin, Andre Choo, Shaul Reuveny,

Viral Infectious Diseases and Gene Expression in Insects

The conventional method of culturing human embryonic stem cells (hESC) is on two-dimensional (2D) surfaces, which is not amenable for scale up to therapeutic quantities in bioreactors. We have developed a facile and robust method for maintaining undifferentiated hESC in three-dimensional (3D) suspension cultures on matrigel-coated microcarriers achieving 2- to 4-fold higher cell densities than those in 2D colony cultures. Stable, continuous propagation of two hESC lines on microcarriers has been demonstrated in conditioned media for 6 months. Microcarrier cultures (MC) were also demonstrated in two serum-free defined media (StemPro and mTeSR1). MC achieved even higher cell concentrations in suspension spinner flasks, thus opening the prospect of propagation in controlled bioreactors.