Methods for making cells with an extra-embryonic endodermal precursor phenotype
View Patent ↗The invention is directed to methods for making cells in vitro that have an extra-embryonic endodermal precursor phenotype. The method involves culturing cells from blastocysts under specific defined conditions.
1. A method to produce extra-embryonic endodermal precursor cells that express extra-embryonic endodermal precursor genes, the method comprising culturing a blastocyst in a culture medium containing platelet-derived growth factor under conditions of reduced oxygen to produce the extra-embryonic endodermal precursor cells wherein the conditions for reduced oxygen include conditions less than about 20%.
2. The method of claim 1 wherein the oxygen concentration is about 10% or less.
3. The method of claim 2 wherein the oxygen concentration is about 5%.
4. The method of claim 1 , wherein the extra-embryonic precursor cells express one or more transcription factors selected from the group consisting of oct4, rex-1, Ecats, Gata4, Gata6, Sox7, Sox17, Foxa2, HNF1Beta, and HNF4alpha.
5. The method of any one of claims 1 - 3 and 4 wherein the blastocysts are cultured without a cellular feeder layer.
6. The method of any one of claims 1 - 3 and 4 , wherein the serum concentration is about 2% to about 5%.
7. The method of any one of claims 1 - 3 and 4 , wherein the serum concentration is about 2%.