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US 12,714,728 B2Granted
Methods for the production of cardiac fibroblasts
AssigneesFUJIFILM Cellular Dynamics, Inc., FUJIFILM Holdings America Corporation
InventorsSarah Himmerich, Nathaniel Beardsley, Annie Armstrong Smelter, Nathan Meyer, Ravi Vaidyanathan, Cara Rieger
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Abstract
Provided herein are methods for producing quiescent cardiac fibroblast cells from pluripotent stem cells. Further provided herein are microtissue compositions comprising the cardiac fibroblast cells with endothelial cells and cardiomyocytes.
Claims32
An in vitro method for producing human pluripotent stem cell (PSC)-derived cardiac fibroblast progenitor cells comprising:
- (a) culturing PSC aggregates in media comprising a Wnt agonist, Activin agonist, and BMP4 for mesoderm induction for about one day;
- (b) further culturing the PSC aggregates in media comprising an Activin agonist and BMP4 and essentially free of a Wnt agonist for about 1-3 days to produce a population of mesoderm progenitor cells that are positive for NCAM and have low expression of CXCR4 (NCAM+/CXCR4low);
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- (c) culturing the mesoderm progenitor cells in media comprising a Wnt inhibitor for about 1-3 days to produce a population of cardiac fibroblast progenitor cells, wherein the cardiac fibroblast progenitor cells comprise epicardial progenitor cells, endothelial fibroblast progenitor cells, and second heart field progenitors.
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Dependencies are inferred from the claim text.