Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells.
Preparation of specific lineages at high purities from embryonic stem (ES) cells requires both selective culture conditions and markers to guide and monitor the differentiation. In this study, we distinguished definitive and visceral endoderm by using a mouse ES cell line that bears the gfp and human IL2R alpha (also known as CD25) marker genes in the goosecoid (Gsc) and Sox17 loci, respectively. This cell line allowed us to monitor the generation of Gsc+ Sox17+ definitive endoderm and Gsc- Sox17+ visceral endoderm and to define culture conditions that differentially induce definitive and visceral endoderm. By comparing the gene expression profiles of definitive and visceral endoderm, we identified seven surface molecules that are expressed differentially in the two populations. One of the seven markers, Cxcr4, to which a monoclonal antibody is available allowed us to monitor and purify the Gsc+ population from genetically unmanipulated ES cells under the condition that selects definitive endoderm.
Yasunaga
Masahiro
M
Basic Research Laboratory, Stem Cell Sciences KK, Kobe, Hyogo 650-0047, Japan.
Tada
Shinsuke
S
Torikai-Nishikawa
Satomi
S
Nakano
Yoko
Y
Okada
Mitsuhiro
M
Jakt
Lars Martin
LM
Nishikawa
Satomi
S
Chiba
Tsutomu
T
Era
Takumi
T
Nishikawa
Shin-Ichi
S
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004707:Endoderm / Q000166:cytology* / Q000502:physiology*; D051379:Mice; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*