	US 11,672,832 B2
	Development of amnion-like tissue from human pluripotent stem cells
Jianping Fu, Ann Arbor, MI (US); Deborah Gumucio, Ann Arbor, MI (US); Yue Shao, Ann Arbor, MI (US); Kenichiro Taniguchi, Ann Arbor, MI (US); Yi Zheng, Ann Arbor, MI (US); and Sajedeh Nasr Esfahani, Ann Arbor, MI (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF MICHIGAN, Ann Arbor, MI (US)
Appl. No. 16/467,154
Filed by THE REGENTS OF THE UNIVERSITY OF MICHIGAN, Ann Arbor, MI (US)
PCT Filed Dec. 8, 2017, PCT No. PCT/US2017/065261 § 371(c)(1), (2) Date Jun. 6, 2019, PCT Pub. No. WO2018/106997, PCT Pub. Date Jun. 14, 2018.
Claims priority of provisional application 62/431,907, filed on Dec. 9, 2016.
Prior Publication US 2019/0321415 A1, Oct. 24, 2019
Int. Cl. A61K 35/545 (2015.01); C12N 5/073 (2010.01); A61K 31/74 (2006.01); A61K 35/50 (2015.01); C12N 5/00 (2006.01); G01N 33/50 (2006.01)

CPC A61K 35/545 (2013.01) [A61K 31/74 (2013.01); A61K 35/50 (2013.01); C12N 5/0068 (2013.01); C12N 5/0605 (2013.01); G01N 33/5044 (2013.01); C12N 2502/45 (2013.01); C12N 2533/30 (2013.01)]

6 Claims

1. A method for preparing amnion-like tissue comprising:

a) contacting cells selected from a group consisting of human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) on a solid support coated with a gel matrix with a thickness of 100 μm, and a Young's bulk modulus of 900 Pa, comprising microposts with a height of 8.4 μm;

b) culturing said hiPSCs or hESCs in a medium comprising Rho-kinase (ROCK) inhibitor Y-2763212 for 24 hours;

c) coating said cells on a said solid support with further gel matrix such that said cells are surrounded by said gel matrix after step b);

d) replacing said Y-2763212 medium with a Y-2763212-free medium, and

e) culturing said hiPSCs or hESCs for 5 days under conditions such that amnion-like tissue is generated, wherein said amnion-like tissue comprises a squamous cyst comprising an epithelium, wherein said squamous cyst does not express NANOG, OCT, or SOX.