US 11,857,697 B2
Compositions and methods for obtaining 3-dimensional lung-like epithelium and related uses thereof
Jason Spence, Ann Arbor, MI (US); and Alyssa Miller, Ann Arbor, MI (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF MICHIGAN, Ann Arbor, MI (US)
Filed by THE REGENTS OF THE UNIVERSITY OF MICHIGAN, Ann Arbor, MI (US)
Filed on May 31, 2018, as Appl. No. 15/994,749.
Claims priority of provisional application 62/513,191, filed on May 31, 2017.
Prior Publication US 2018/0344901 A1, Dec. 6, 2018
Int. Cl. C12N 5/071 (2010.01); C12N 5/00 (2006.01); A61K 35/42 (2015.01); A61L 27/38 (2006.01); C12N 5/074 (2010.01); C12N 5/0735 (2010.01)
CPC A61L 27/3895 (2013.01) [A61K 35/42 (2013.01); A61L 27/3804 (2013.01); A61L 27/3813 (2013.01); A61L 27/3834 (2013.01); A61L 27/3839 (2013.01); A61L 27/3869 (2013.01); C12N 5/0018 (2013.01); C12N 5/0062 (2013.01); C12N 5/0606 (2013.01); C12N 5/0607 (2013.01); C12N 5/0688 (2013.01); C12N 5/0689 (2013.01); A61L 2430/22 (2013.01); C12N 2501/113 (2013.01); C12N 2501/115 (2013.01); C12N 2501/117 (2013.01); C12N 2501/119 (2013.01); C12N 2501/155 (2013.01); C12N 2501/385 (2013.01); C12N 2501/415 (2013.01); C12N 2501/999 (2013.01); C12N 2506/02 (2013.01); C12N 2506/03 (2013.01); C12N 2506/27 (2013.01); C12N 2506/45 (2013.01); C12N 2513/00 (2013.01)] 11 Claims
 
1. A method, comprising:
culturing in vitro for approximately forty days cells from NKX2.1+ ventral-anterior foregut spheroid tissue derived from pluripotent stem cells, wherein the culturing results in differentiation of the cells from the NKX2.1+ ventral-anterior foregut spheroid tissue derived from pluripotent stem cells into tissue comprising 3-dimensional lung-like epithelium, wherein the culturing comprises activating the FGF signaling pathway, the retinoic acid signaling pathway, and the Wnt signaling pathway; and
obtaining 3-dimensional lung tissue from the cultured tissue comprising 3-dimensional lung-like epithelium, wherein the obtained 3-dimensional lung tissue comprises a 3-dimensional lung tissue peripheral region and a 3-dimenstional lung tissue internal region,
wherein the 3-dimensional lung tissue peripheral region comprises cells expressing SOX9 protein and SOX2 protein,
wherein the 3-dimensional lung tissue internal region comprises cells expressing SOX2 protein, SCGB1A1 protein, and MUC5AC, wherein the 3-dimensional lung tissue internal region does not comprise cells expressing SOX9 protein.