	US 12,297,452 B2
	Methods or generating T-cells from stem cells and immunotherapeutic methods using the T-cells
Gay M. Crooks, Sherman Oaks, CA (US); Amélie Montel-Hagen, Los Angeles, CA (US); and Christopher Seet, Santa Monica, CA (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA (US)
Filed by The Regents of the University of California, Oakland, CA (US)
Filed on Sep. 17, 2021, as Appl. No. 17/478,875.
Application 17/478,875 is a continuation of application No. 15/772,224, granted, now 11,154,573, previously published as PCT/US2016/059375, filed on Oct. 28, 2016.
Claims priority of provisional application 62/359,456, filed on Jul. 7, 2016.
Claims priority of provisional application 62/265,204, filed on Dec. 9, 2015.
Claims priority of provisional application 62/248,931, filed on Oct. 30, 2015.
Prior Publication US 2022/0096553 A1, Mar. 31, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/00 (2006.01); A61K 39/00 (2006.01); A61P 35/00 (2006.01); C07K 14/705 (2006.01); C12N 5/0735 (2010.01); C12N 5/074 (2010.01); C12N 5/077 (2010.01); C12N 5/0775 (2010.01); C12N 5/0783 (2010.01)

CPC C12N 5/0062 (2013.01) [A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/4632 (2023.05); A61K 39/4644 (2023.05); A61P 35/00 (2018.01); C07K 14/705 (2013.01); C12N 5/00 (2013.01); C12N 5/0037 (2013.01); C12N 5/0606 (2013.01); C12N 5/0636 (2013.01); C12N 5/0663 (2013.01); C12N 5/0669 (2013.01); C12N 5/0696 (2013.01); C07K 2317/622 (2013.01); C07K 2319/03 (2013.01); C12N 2500/22 (2013.01); C12N 2500/24 (2013.01); C12N 2500/32 (2013.01); C12N 2500/38 (2013.01); C12N 2500/90 (2013.01); C12N 2501/125 (2013.01); C12N 2501/145 (2013.01); C12N 2501/15 (2013.01); C12N 2501/2302 (2013.01); C12N 2501/2304 (2013.01); C12N 2501/2306 (2013.01); C12N 2501/2307 (2013.01); C12N 2501/2315 (2013.01); C12N 2501/2321 (2013.01); C12N 2501/25 (2013.01)]

10 Claims

1. A method for preparing a composition of mature CD4⁺ CD8⁻, or CD8ab⁺ CD4⁻ T cells from stem or progenitor cells, the method comprising culturing a three-dimensional (3D) cell aggregate comprising:

a) a cell line of stromal cells that express a Notch ligand; and

b) a population of stem or progenitor cells generated in vitro from embryonic stem cells (ESCs), induced pluripotent stem cells (iPSC), or human embryonic mesodermal progenitor cells;

wherein the 3D cell aggregate is cultured in a serum-free medium for a time period sufficient for in vitro differentiation of the stem or progenitor cells to mature T cells, wherein the 3D cell aggregate does not comprise an exogenous matrix or a scaffold.