Mesoderm and definitive endoderm cell populations
The present invention provides cell populations that are enriched for mesendoderm and mesoderm, and cell populations that are enriched for endoderm. The cell populations of the invention are useful for generating cells for cell replacement therapy. The present invention further provides a method of generating hepatocytes, cell populations enriched for hepatocytes, and a method of hepatocyte replacement therapy.
1. A method of making a cell population enriched for endoderm cells comprising culturing pluripotent stem cells in the presence of activin or nodal, and Wnt, for a time sufficient to provide brach + /HNF3β + cells, and culturing said brach + /HNF3β + cells in the presence of activin and an inhibitor of Wnt signalling for a time sufficient to provide a cell population enriched for endoderm cells.
2. The method of claim 1 wherein the stem cells are cultured in the absence of serum.
3. The method of claim 1 wherein the brach + /HNF3β + cells are cultured in the absence of serum.
4. The method of claim 1 wherein the stem cells are human embryonic stem cells.
5. The method of claim 1 wherein Wnt is Wnt 3d and the inhibitor of Wnt signaling is DKK-1.
6. The method of claim 1 wherein activin is present at a concentration of at least about 30 ng/ml.