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  • Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines.

Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines.

Tissue engineering. Part A (2009-01-10)
Heidi Hakala, Kristiina Rajala, Marisa Ojala, Sarita Panula, Sami Areva, Minna Kellomäki, Riitta Suuronen, Heli Skottman
RESUMEN

Long-term in vitro culture of undifferentiated human embryonic stem cells (hESCs) traditionally requires a fibroblast feeder cell layer. Using feeder cells in hESC cultures is highly laborious and limits large-scale hESC production for potential application in regenerative medicine. Replacing feeder cells with defined human extracellular matrix (ECM) components or synthetic biomaterials would be ideal for large-scale production of clinical-grade hESCs. We tested and compared different feeder cell-free hESC culture methods based on different human ECM proteins, human and animal sera matrices, and a Matrigel matrix. Also selected biomaterials were tested for feeder cell-free propagation of undifferentiated hESCs. The matrices were tested together with conventional and modified hESC culture media, human foreskin fibroblast-conditioned culture medium, chemically defined medium, TeSR1, and modified TeSR1 media. The results showed the undefined, xenogeneic Matrigel to be a superior matrix for hESC culture compared with the purified human ECM proteins, serum matrices, and the biomaterials tested. A long-term, feeder cell-free culture system was successful on Matrigel in combination with mTeSR1 culture medium, but a xeno-free, fully defined, and reproducible feeder cell-free hESC culture method still remains to be developed.

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Fibronectina, cell culture derived, ~0.5 mg/mL, sterile-filtered, BioReagent, suitable for cell culture