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Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells

Unger, Christian
Gao, Shuping
Jaconi, Marisa
Bergstrom, Rosita
Holm, Frida
Galan, Amparo
Sanchez, Eva
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Published in Human reproduction. 2009, vol. 24, no. 10, p. 2567-81
Abstract BACKGROUND: Feeder cells are frequently used for the early-stage of derivation and culture of human embryonic stem cell (hESC) lines. METHODS: We established a conditionally immortalized human foreskin fibroblast line that secreted basic fibroblast growth factor (bFGF). These cells were used as feeder cells for hESC culture and induced pluripotent stem (iPS) cell derivation and expansion. This conditional immortalization was performed using lentiviral vector (LV) mediated transduction of Bmi-1 and human telomerase reverse transcriptase genes and the resulting cell line was further modified by LV-mediated transduction of a secreted form of bFGF gene product. Three different laboratories have tested whether this feeder cell line could support the maintenance of four different hESC lines. RESULTS: Immortalized fibroblasts secreting stable amounts of bFGF supported the growth of all hESC lines, which remained pluripotent and had a normal karyotype for at least 10 passages. Even at high passage (p56), these modified cells, when used as feeders, could support iPS derivation and propagation. Derived iPS cells expressed pluripotency markers, had hESC morphology and produced tissue components of the three germ layers when differentiated in vitro. CONCLUSION: These modified fibroblasts are useful as a genetically-defined feeder cell line for reproducible and cost-effective culture of both hESC and iPS cells.
Keywords AnimalsCell Culture TechniquesCell DifferentiationCell LineCell ProliferationCoculture TechniquesEmbryonic Stem Cells/physiologyFibroblasts/cytologyHumansKaryotypingLentivirus/geneticsMicePluripotent Stem Cells/physiology
PMID: 19556288
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Research groups Laboratoire de biologie des tumeurs gynécologiques et du développement (927)
Martinez De Tejada Begona (272)
Mécanismes de différentiation cellulaire des cellules cardiaques (536)
Progéniteurs neuronaux (141)
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UNGER, Christian et al. Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells. In: Human Reproduction, 2009, vol. 24, n° 10, p. 2567-81. doi: 10.1093/humrep/dep232 https://archive-ouverte.unige.ch/unige:5295

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Deposited on : 2010-03-02

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