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Stem Cell Res. 2009 May;2(3):219-30. doi: 10.1016/j.scr.2009.02.005. Epub 2009 Mar 4.

Long-term microcarrier suspension cultures of human embryonic stem cells.

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Stem Cell Group, Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology, and Research), 20 Biopolis Way, No. 06-01 Centros, Singapore 138668, Singapore.


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.

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