Differentiation of pluripotent stem cells and cardiac progenitor cells into striated cardiomyocyte fibers using laminins LN-511, LN-521 and LN-221
AssigneesNational University of Singapore, Biolamina AB
InventorsKarl Tryggvason, Yan Wen Yap, Kristian Tryggvason, Yi Sun
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Abstract
The present disclosure describes methods of differentiating cardiomyocyte progenitor cells and mature cardiomyocyte cells from pluripotent stem cells. The methods may include differentiating pluripotent stems cells on a substrate including (i) laminin-511 or 521 and (ii) laminin-221. The cardiomyocyte progenitor cells and mature cardiomyocyte cells produced by the methods may form a human heart muscle cell line for use in regenerative cardiology. Also described are methods of identifying functional cardiomyocyte progenitor cells and their use in therapeutic applications.
Claims6
A method for treating injured human cardiac muscle, comprising:
- (a) culturing human pluripotent stem cells on a culture plate coated with a substrate in a medium containing basic fibroblast growth factor (bFGF), wherein the substrate comprises a mixture of two laminins consisting of (i) recombinant LN-521 or recombinant LN-511 and (ii) recombinant LN-221;
- (b) culturing the pluripotent stem cells on the substrate in the presence of a differentiation medium containing an effective amount of a GSK-3 inhibitor to stimulate Wnt signaling and obtain brachyury positive cells;
- (c) culturing the brachyury positive cells from step (b) on the substrate in a differentiation medium containing an effective amount of a Wnt inhibitor to promote cardiac mesodermal specification;
- (d) removing the differentiation medium containing the Wnt inhibitor from step (c) and replacing with a basal medium to culture the cells on the substrate to produce human cardiomyocyte progenitor cells that express Islet-1 and NKX2.5 transcription factor;
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- (e) transplanting the human cardiomyocyte progenitor cells into the injured human cardiac muscle.
Dependencies are inferred from the claim text.