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

In: Human Reproduction, 2009, vol. 24, no. 10, p. 2567-2581

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    Title
    • Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells
    Author
    • Unger, Christian. Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
    • Gao, Shuping. Laboratory of Stem Cell Research, Geneva University Hospitals, 30, Bld de la Cluse, CH-1211 Geneva, Switzerland
    • Cohen, Marie. Laboratory of Stem Cell Research, Geneva University Hospitals, 30, Bld de la Cluse, CH-1211 Geneva, Switzerland
    • Jaconi, Marisa. Department of Pathology and Immunology, Faculty of Medicine, Geneva University, Geneva, Switzerland
    • Bergstrom, Rosita. Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
    • Holm, Frida. Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
    • Galan, Amparo. Spanish Stem Cell Bank, Comunidad Valenciana Node, Prince Felipe Research Centre (CIPF), Valencia, Spain
    • Sanchez, Eva. Spanish Stem Cell Bank, Comunidad Valenciana Node, Prince Felipe Research Centre (CIPF), Valencia, Spain
    • Irion, Olivier. Department of Obstetrics and Gynaecology, Geneva University Hospitals, Geneva, Switzerland
    • Dubuisson, Jean Bernard. Department of Obstetrics and Gynaecology, Geneva University Hospitals, Geneva, Switzerland
    • Giry-Laterriere, Marc. Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
    • Salmon, Patrick. Department of Neurosciences, Faculty of Medicine, Geneva University, Geneva, Switzerland
    • Simon, Carlos. Spanish Stem Cell Bank, Comunidad Valenciana Node, Prince Felipe Research Centre (CIPF), Valencia, Spain
    • Hovatta, Outi. Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
    • Feki, Anis. Laboratory of Stem Cell Research, Geneva University Hospitals, 30, Bld de la Cluse, CH-1211 Geneva, Switzerland
    Document Type
    • Postprint
    Language
    • English
    Published in
    • Human Reproduction, 2009, vol. 24, no. 10, p. 2567-2581. Oxford University Press
    Other Version
    OAI-PMH Identifier
    • oai:doc.rero.ch:304688
    Summary
    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