Long-term microcarrier suspension cultures of human embryonic stem cells.
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.
Oh
Steve K W
SK
Stem Cell Group, Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology, and Research), 20 Biopolis Way, No. 06-01 Centros, Singapore 138668, Singapore. steve_oh@bti.a-star.edu.sg
Chen
Allen K
AK
Mok
Yanglin
Y
Chen
Xiaoli
X
Lim
U-Ming
UM
Chin
Angela
A
Choo
Andre B H
AB
Reuveny
Shaul
S
D019149:Bioreactors; D018929:Cell Culture Techniques; D003094:Collagen / Q000737:chemistry; D016895:Culture Media, Serum-Free; D004338:Drug Combinations; D004475:Ectoderm / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004707:Endoderm / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D007797:Laminin / Q000737:chemistry; D008648:Mesoderm / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry
D016895:Culture Media, Serum-Free; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen