Granted Patent
B2
US 12,692,476 · App. 18/041,174 · Granted Jul 28, 2026
Compositions and methods for embryonic stem cell expansion
Inventors:
Lilach Alon (Kfar Yona, IL); Rami Skaliter (Ness Ziona, IL); Ravid Tikotzki (Rishon Lezion, IL); Dana Hayoun Neeman (Jerusalem, IL); Ofer Wiser (Jerusalem, IL)
Assignee:
LINEAGE CELL THERAPEUTICS, INC.
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Abstract
Provided herein are methods and compositions for expansion of human embryonic stem with suspendable expansion complexes including microcarriers.
Claims (18)
1 . A method for expanding and maintaining human stem cells in an undifferentiated, pluripotent state, the method comprising the steps of (a) simultaneously combining human stem cells, human laminin 511 E8 fragment, and polystyrene microcarriers in growth medium to form a suspendable expansion complex, and (b) culturing the suspendable expansion complex for a period of time, thereby producing cultured cells.
2 . The method of claim 1 , wherein the polystyrene microcarriers comprise one or more of polystyrene, cross-linked dextran, magnetic particles, microchips, cellulose, hydroxylated methacrylate, collagen, gelatin, plastic, glass, ceramic, silicone, or a combination thereof.
3 . The method of claim 1 , wherein the polystyrene microcarriers are spherical, smooth, macroporous, rod-shaped, or a combination thereof.
4 . The method of claim 1 , wherein the polystyrene microcarriers are not coated.
5 . The method of claim 1 , wherein the polystyrene microcarriers are coupled with protamine or polylysine.
6 . The method of claim 1 , wherein the polystyrene microcarriers are neutral or negatively charged.
7 . The method of claim 1 , wherein the polystyrene microcarriers are hydrophilic.
8 . The method of claim 1 , wherein the suspendable expansion complex is cultured for at least about one day.
9 . The method of claim 8 , wherein the suspendable expansion complex is cultured from about one day to about fourteen days.
10 . The method of claim 1 , wherein the cultured cells of the suspendable expansion complex are harvested and expanded further by repeating steps (a) and (b).
11 . The method of claim 1 , wherein the cultured cells of the suspendable expansion complex are harvested and further differentiated.
12 . The method of claim 11 , wherein the cultured cells of the suspendable expansion complex are further differentiated by changing the growth medium.
13 . The method of claim 1 , wherein the cultured cells of the suspendable expansion complex remain undifferentiated and pluripotent.
14 . The method of claim 1 , wherein at least about 80% of the cultured cells express each of SSEA-5 and TRA-1-60.
15 . The method of claim 1 , wherein at least about 70% of the cultured cells express each of Oct-4 and Nanog.
16 . The method of claim 1 , wherein at least about 80% of the cultured cells express SSEA-5, at least about 80% of the cultured cells express TRA-1-60, at least about 70% of the cultured cells express Oct-4, and at least about 70% of the cultured cells express Nanog.
17 . The method of claim 1 , wherein the human stem cells are induced pluripotent stem cells (iPSCs) or human embryonic stem cells.
18 . The method of claim 1 , wherein the polystyrene microcarriers are surface treated with infused oxygen.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 28, 2023
From: HAYOUN NEEMAN, DANA; WISER, OFER; ALON, LILACH; TIKOTZKI, RAVID; SKALITER, RAMI
To: LINEAGE CELL THERAPEUTICS, INC.
Reel/Frame 064102/0596 →
Continuity (2)
Provisional Application
63063942
· Aug 10, 2020
Related Publication
20230313127A1
· Oct 5, 2023
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