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US 9,752,118 B2Granted
Method of directed differentiation producing corneal endothelial cells from neural crest stem cells by PDGFB and DKK2, compositions thereof, and uses thereof
AssigneeAstellas Institute for Regenerative Medicine
InventorsKathryn L. McCabe, Shi-Jiang Lu, Robert P. Lanza
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Abstract
This disclosure generally relates to cell-based therapies for treatment of visual disorders, including disorders of the cornea. Methods are exemplified for directed differentiation of corneal cells from stem cells. Compositions of corneal endothelial cells and uses thereof are also provided. Exemplary compositions exhibit improved cell density and/or more “youthful” gene expression relative to cells obtained from donated tissue.
Claims9
A method of producing corneal endothelial cells (CECs), comprising
- contacting embryonic stem (ES) cells with Noggin and 4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]benzamide (SB431542) to induce differentiation of the ES cells into neural crest stem cells (NCSCs);
- and
- contacting the NCSCs with dickkopf-related protein 2 (DKK2), and platelet-derived growth factor subunit B (PDGFB), to induce differentiation of the NCSCs into CECs that express Na+/K+ ATPase, ZO-1, KLF13, AQP1, Collagen VIII, SLC16A3, CFTR, NBC1, Carbonic anhydrase II (CA2), Anion exchanger 2 (AE2/SCL4A2), Solute Carrier Family 16 A1 (SCL16A1), SCL16A3, Carbonic anhydrase 12 (CA12), Carbonic Anhydrase 4 (CA4), cystic fibrosis transmembrane conductance regulator (CFTR) and transcription factor FoxC1.
Dependencies are inferred from the claim text.