US 12,467,036 B2
Methods for culturing and differentiating pluripotent cells into progenitor or mature muscle cells and composition comprising said cells
Iftach Nachman, Tel Aviv (IL); and Gaya Savyon, Tel Aviv (IL)
Assigned to RAMOT AT TEL AVIV UNIVERSITY LTD., Tel Aviv (IL)
Appl. No. 18/278,100
Filed by RAMOT AT TEL AVIV UNIVERSITY LTD., Tel Aviv (IL)
PCT Filed Mar. 20, 2022, PCT No. PCT/IL2022/050310
§ 371(c)(1), (2) Date Aug. 21, 2023,
PCT Pub. No. WO2022/201147, PCT Pub. Date Sep. 29, 2022.
Claims priority of provisional application 63/164,047, filed on Mar. 22, 2021.
Prior Publication US 2024/0052315 A1, Feb. 15, 2024
Int. Cl. C12N 5/077 (2010.01)
CPC C12N 5/0658 (2013.01) [C12N 2500/25 (2013.01); C12N 2501/105 (2013.01); C12N 2501/115 (2013.01); C12N 2501/12 (2013.01); C12N 2501/155 (2013.01); C12N 2501/415 (2013.01); C12N 2506/02 (2013.01)] 12 Claims
 
1. A method for generating somites, the method comprising:
(a) suspending embryonic stem cells in culture medium to obtain spheric aggregates;
(b) adding to the culture medium at least one Wnt activator to obtain aggregates comprising mesodermal and endodermal cells;
(c) removing the Wnt activator;
(d) identifying pre-somitic mesoderm (PSM) cells having a Mesogenin1 (Msgn1) marker in the aggregates comprising the mesodermal and endodermal cells, and adding extracellular matrix or components thereof when said PSM cells accumulate at poles of the aggregate, to obtain aggregates comprising paraxial mesoderm; and
(e) incubating the aggregates of step (d), for 10 to 48 hours, thereby obtaining aggregates comprising a plurality of somites,
wherein said embryonic stem cells are selected from the group consisting of human and mouse embryonic stem cells.