Development of definitive endoderm from embryonic stem cells in culture.
The cellular and molecular events regulating the induction and tissue-specific differentiation of endoderm are central to our understanding of the development and function of many organ systems. To define and characterize key components in this process, we have investigated the potential of embryonic stem (ES) cells to generate endoderm following their differentiation to embryoid bodies (EBs) in culture. We found that endoderm can be induced in EBs, either by limited exposure to serum or by culturing in the presence of activin A (activin) under serum-free conditions. By using an ES cell line with the green fluorescent protein (GFP) cDNA targeted to the brachyury locus, we demonstrate that endoderm develops from a brachyury(+) population that also displays mesoderm potential. Transplantation of cells generated from activin-induced brachyury(+) cells to the kidney capsule of recipient mice resulted in the development of endoderm-derived structures. These findings demonstrate that ES cells can generate endoderm in culture and, as such, establish this differentiation system as a unique murine model for studying the development and specification of this germ layer.
Kubo
Atsushi
A
The Carl C. Icahn Center for Gene Therapy and Molecular Medicine, Mount Sinai School of Medicine, New York, NY 10029, USA.
Shinozaki
Katsunori
K
Shannon
John M
JM
Kouskoff
Valerie
V
Kennedy
Marion
M
Woo
Savio
S
Fehling
Hans Joerg
HJ
Keller
Gordon
G
D028341:Activins / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage; D017690:Cell Transplantation; D016895:Culture Media, Serum-Free; D004627:Embryonic Induction / Q000502:physiology*; D004707:Endoderm / Q000166:cytology / Q000502:physiology*; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology; D006801:Humans; D028322:Inhibin-beta Subunits / Q000378:metabolism; D007668:Kidney / Q000033:anatomy & histology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000502:physiology; D051379:Mice; D016513:Mice, SCID; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D000098908:Brachyury Protein
D028341:Activins; D016895:Culture Media, Serum-Free; D005326:Fetal Proteins; D049452:Green Fluorescent Proteins; D028322:Inhibin-beta Subunits; D008164:Luminescent Proteins; D020825:T-Box Domain Proteins; D000098908:Brachyury Protein; C422405:activin A