US 12,371,669 B2
Kidney organoids and method for producing the same
Dong Sung Kim, Pohang-si (KR); Tae Eun Park, Ulsan (KR); Hyeon Ji Lim, Ulsan (KR); and Do Hui Kim, Incheon (KR)
Assigned to POSTECH RESEARCH AND BUSINESS DEVELOPMENT FOUNDATION, Pohang-si (KR); and UNIST(ULSAN NATIONAL INSTITUTE OF SCIENCE AND TECHNOLOGY), Ulsan (KR)
Filed by POSTECH Research and Business Development Foundation, Pohang-si (KR); and UNIST(ULSAN NATIONAL INSTITUTE OF SCIENCE AND TECHNOLOGY), Ulsan (KR)
Filed on Nov. 4, 2022, as Appl. No. 17/980,820.
Claims priority of application No. 10-2021-0153107 (KR), filed on Nov. 9, 2021; and application No. 10-2022-0120733 (KR), filed on Sep. 23, 2022.
Prior Publication US 2023/0142476 A1, May 11, 2023
Int. Cl. C12N 5/071 (2010.01)
CPC C12N 5/0697 (2013.01) [C12N 2500/02 (2013.01); C12N 2501/119 (2013.01); C12N 2501/16 (2013.01); C12N 2506/45 (2013.01); C12N 2513/00 (2013.01)] 4 Claims
 
1. A method for producing human kidney organoids comprising:
(A) differentiating human pluripotent stem cells into metanephric mesenchyme cells by:
(i) culturing the human pluripotent stem cells in a first medium containing a GSK-3β inhibitor and a BMP4 inhibitor on Day 0,
(ii) culturing the human pluripotent stem cells in a second medium containing Activin A on day 4,
(iii) culturing the human pluripotent stem cells in a third medium containing 10 ng/ml of FGF9 on day 7;
(B) forming metanephric mesenchyme cell aggregates by culturing the metanephric mesenchyme cells in a fourth medium containing a GSK-3p inhibitor and 10 ng/ml of FGF9 on Day 9; and
(C) differentiating the metanephric mesenchyme cell aggregates into kidney organoids in a fifth medium containing FBS on day 14 for 7 to 100 days and culturing the metanephric mesenchyme cells in a sixth medium containing 25 ng/ml to 50 ng/ml of FGF9 on Day 11 to form kidney organoids;
wherein steps (B) and (C) are performed in a porous microwell formed of a permeable porous membrane comprising at least one concave portion,
wherein the at least one concave portion is formed at the bottom and an opening is formed at the top of the microwell,
wherein the porous microwells facilitate supply of nutrients and growth factors and removal of metabolites.