Journal Article

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

Christian Unger, Shuping Gao, Marie Cohen, Marisa Jaconi, Rosita Bergstrom, Frida Holm, Amparo Galan, Eva Sanchez, Olivier Irion, Jean Bernard Dubuisson, Marc Giry-Laterriere, Patrick Salmon, Carlos Simon, Outi Hovatta and Anis Feki

in Human Reproduction

Published on behalf of European Society of Human Reproduction and Embryology

Volume 24, issue 10, pages 2567-2581
Published in print October 2009 | ISSN: 0268-1161
Published online June 2009 | e-ISSN: 1460-2350 | DOI: http://dx.doi.org/10.1093/humrep/dep232
Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells

Show Summary Details

Preview

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: feeder cells; human embryonic stem cells; induced pluripotent stem cells; human fibroblasts; immortalization

Journal Article.  6493 words.  Illustrated.

Subjects: Reproductive Medicine

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.