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


CONCLUSIONS
These modified fibroblasts are useful as a genetically-defined feeder cell line for reproducible and cost-effective culture of both hESC and iPS cells.
Unger
Christian
C
Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
Gao
Shuping
S
Cohen
Marie
M
Jaconi
Marisa
M
Bergstrom
Rosita
R
Holm
Frida
F
Galan
Amparo
A
Sanchez
Eva
E
Irion
Olivier
O
Dubuisson
Jean Bernard
JB
Giry-Laterriere
Marc
M
Salmon
Patrick
P
Simon
Carlos
C
Hovatta
Outi
O
Feki
Anis
A
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D007621:Karyotyping; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D039904:Pluripotent Stem Cells / Q000502:physiology*