Generation of pancreatic endoderm from stem cell derived definitive endoderm
AssigneeAspect Biosystems Ltd.
InventorsDorthe Roenn Petersen, Christian Honore
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Abstract
The present invention relates to methods of efficiently generating pancreatic endoderm from human pluripotent stem (PS) cell derived human definitive endoderm. The present invention also relates to pancreatic endoderm cells obtained by the methods of the invention. Finally, the present invention relates to culture medium and composition comprising a RAR antagonist and uses of said RAR antagonist in the induction of pancreatic endoderm cells. The present invention provides a more homogenous and synchronised pancreatic cell population, with increased efficiency.
Claims44
A method of differentiating definitive endoderm (DE) cells derived from human pluripotent stem cells into pancreatic endoderm precursors comprising culturing the DE cells in a culture medium comprising a retinoic acid receptor (RAR) antagonist.
A synchronised population of pancreatic endoderm cells derived from human pluripotent stem cells, wherein at least 70% of the PE cells are PDX1+/NKX6.1+ double positive.
12-13. (canceled)
A method of differentiating definitive endoderm (DE) cells derived from human pluripotent stem cells into pancreatic endoderm (PE) cells, comprising
- culturing the DE cells in a culture medium comprising a retinoic acid receptor (RAR) antagonist to produce pancreatic endoderm precursors, and
- culturing the pancreatic endoderm precursors in a cell culture medium comprising a RAR agonist to produce the PE cells;
- wherein the PE cells are PDX1+/NKX6.1+ double positive;
- wherein the RAR antagonist is selected from the group consisting of AGN193109, AGN 194431, AGN 194301, SR 11335, BMS 453, BMS 195614, LE 135, LG 100815, MM11253, CD 2665, and ER 50891;
- wherein the RAR agonist is selected from the group consisting of AM580, All-trans retinoic acid, 9-cis retinoic acid, AC 261066, AC 55649, Adapalene, AM 80, BMS 753, BMS 961, CD 1530, CD 2314, CD 437, Ch 55, Isotretinoin, Tazarotene, TTNTB, and EC19;
- and
- wherein the concentration of the RAR antagonist is in a range of 0.5-100 μM.
Dependencies are inferred from the claim text.