Unformatted text preview: quality of the cells since
there is no contact of blastocyst with animal-derived
pronase, antibodies, and complement factors. The latter
procedure has been recently adapted for the establishment of new hESC line by Genbacev et al. (2005).
We have also derived our latest ﬁve hESC lines
mechanically (Fig. 1). Feeder cells in human embryonic stem cell cultures
The establishment of hESC lines was ﬁrst described by
using mouse embryonic ﬁbroblasts (MEFs) as feeder cells
and fetal bovine serum (FBS)-containing culture medium
for both feeder cells and hESC (Thomson et al. 1998).
For replacing animal cells, various alternative human cell lines have been successfully tested as feeder cells for hESC.
Table 2 summarizes the sources of human feeders that have
been validated for culture of undifferentiated hESC.
Richards and co-workers have studied different kinds
of human cells, such as human fetal muscle and skin,
human adult Fallopian tube epithelial feeders, and adult
muscle cells as feeders for hESC (Richards et al. 2002,
2003). A ﬁnding from their results was that for some
reason MEF cells seemed to support hESC growth better
than human feeders. Our group has derived and cultured
all our 25 h ESC lines using commercially available
human foreskin ﬁbroblasts (hFF) as feeder cells (Hovatta
et al. 2003, Inzunza et al. 2005). In our cultures, hFF
seem to support the derivation and undifferentiated
growth of hESC well. On the other hand, we have not
compared the growth rates of hESC cultured with hFF or
MEFs as feeder cells. The commercially available hFF
lines (and other reported lines) have been derived
and cultured in FBS-containing medium which often
has batch-to-batch variation causing variation to culture
conditions. Also, human adult marrow stroma cells
for hESC maintenance (Cheng et al. 2003), and uterine
endometrium cells (Lee et al. 2005) as well as human
placenta ﬁbroblasts (Genbacev et al. 2005) for derivation
and maintenance of hESC have been used, but all
these feeder cells have been cultured using FBS in
culture medium. Table 2 Human cells used as feeder cells for human embryonic stem cell cultures.
Feeder cell source
Adult marrow stroma
Adult fallopian tube
Adult muscle Feeder cell medium composition Derivation (D)/maintenance (M) FBS/HS
Amit et al. (2003)
Hovatta et al. (2003)
Inzunza et al. (2005)
Genbacev et al. (2005)
Lee et al. (2005)
Cheng et al. (2003)
Xu et al. (2004)
Stojkovic et al. (2005b)
Wang et al. (2005c)
Richards et al. (2002)
Richards et al. (2003) FBS, fetal bovine serum; HS, human serum; hESC, human embryonic stem cell.
Reproduction (2006) 132 691–698 www.reproduction-online.org Culture conditions for hESCs 693 Table 3 Feeder-free culture methods established for human embryonic stem cells.
maintenance (M) Coating ma...
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