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Fig. 6. Isolation of mesendoderm cells and endoderm cells from genetically
unmanipulated ES cells. (A) Purification of
ECD+
R+ mesendoderm cells. EB5 unmodified ES cells
were cultured in SFM containing activin. On day 4,
ECD+R+ cells were purified by FACS and
re-cultured for 2 days. Both ECD- and ECD+ populations
(labelled A and B, respectively) derived from day 4
ECD+R+ cells were sorted again by FACS for
further analyses. (B) The expression of mesoderm and endoderm markers in
ECD- and ECD+ populations (labelled A and B,
respectively). A set of endoderm markers such as Foxa2 and
Sox17 was exclusively expressed in B, whereas the expression of
mesoderm markers such as ßR and Vegfr2 was detected
only in A. (C) The morphology of endoderm cell line derived from wild-type ES
cells. Epithelial-like morphology with clear cell-cell boundary is present.
This morphology is similar to endoderm cell lines generated from
Gsc+ECD+ cells (Fig.
4B).
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