Patent ID: 11918702
Assignee: THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
Field: Measurement (Instruments)
Classification: CPC A  C  G | IPC A  C  G

Claim 0:
1. A method for making a branched lung bud organoid (BLBO) from mammalian pluripotent stem cells in a 3D matrix, comprising:
a) selecting at least one lung bud organoid (LBO) with folding structures from anterior foregut endoderm cells, wherein the anterior foregut endoderm cells have been produced from the pluripotent stem cells, wherein the at least one LBO comprises lung epithelial and mesenchymal progenitors, and
b) embedding the at least one LBO within a 3D matrix and culturing the at least one LBO in the presence of a branching medium that induces branching for a duration of time and under conditions that permit at least one branched LBO (BLBO) to form from the at least one LBO, wherein the branching medium comprises CHIR99021, FGF7, FGF10, BMP4 and retinoic acid, wherein the culturing in step b) is conducted for at least 1 week,
wherein embedding in step b) comprises (i) solidifying a first amount of the 3D matrix in a container to form a lower portion of solidified 3D matrix, (ii) solidifying a mixture of the at least one LBO with folding structures and a second amount of the 3D matrix on top of the lower portion to form a solidified center portion; and (iii) solidifying a third amount of the 3D matrix on the solidified center portion to form a top portion,
wherein the second amount of the 3D matrix has a concentration of about 100%, wherein the 3D matrix is a solubilized basement membrane preparation from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma.