Long-term microcarrier suspension cultures of human embryonic stem cells

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

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bioreactors
  • Cell Culture Techniques*
  • Collagen / chemistry
  • Culture Media, Serum-Free
  • Drug Combinations
  • Ectoderm / metabolism
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Endoderm / metabolism
  • Humans
  • Karyotyping
  • Laminin / chemistry
  • Mesoderm / metabolism
  • Proteoglycans / chemistry

Substances

  • Culture Media, Serum-Free
  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen