CPC C12N 5/0657 (2013.01) [C12N 2500/38 (2013.01); C12N 2501/155 (2013.01); C12N 2501/999 (2013.01); C12N 2506/45 (2013.01)] | 13 Claims |
1. An in vitro method of differentiating human pluripotent stem cells into mature atrial cardiomyocytes expressing COUPTFII and COUPTFI, the method comprising the steps of:
a) culturing the human pluripotent stem cells in an aqueous media containing 1-7 μM of a Wnt-signaling agonist and/or a BMP-signaling agonist for a period of about 24-54 hours;
b) culturing the Wnt-signaling agonist and/or BMP-signaling agonist-treated cells obtained from step (a) in an aqueous media containing 0.1-10 μM of a Wnt-signaling antagonist for a period of about 24-54 hours,
wherein during step b) the Wnt-signaling agonist and/or BMP-signaling agonist-treated cells are provided with 2-10 μM of retinoic acid or all-trans retinoic acid within the first 72 hours of step a) and b) and wherein the cells are treated with retinoic acid or all-trans retinoic acid for a period of at least 24 hours; and
c) replacing the aqueous medium of the cells obtained after step (b) with an aqueous medium devoid of the Wnt-signaling agonist and/or BMP-signaling agonist and devoid of the Wnt-signaling antagonist such that mature atrial cardiomyocytes expressing COUPTFII and COUPTFI are obtained.
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