Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines.
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.
Hakala
Heidi
H
REGEA, Institute for Regenerative Medicine, University of Tampere, Tampere, Finland. heidi.minna.hakala@regea.fi
Rajala
Kristiina
K
Ojala
Marisa
M
Panula
Sarita
S
Areva
Sami
S
Kellomäki
Minna
M
Suuronen
Riitta
R
Skottman
Heli
H
D001672:Biocompatible Materials / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D002474:Cell-Free System; D003094:Collagen / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D044967:Serum
D001672:Biocompatible Materials; D004338:Drug Combinations; D007797:Laminin; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen