Granted Patent B2

US 12,421,499 · App. 17/054,633 · Granted Sep 23, 2025

Inventors: Shuibing Chen (Pelham, NY); Sadaf Amin (New York, NY)

Assignee: Cornell University

C12N5/0676C12N2510/00

Abstract

Claims (18)

1. A method of generating an enriched population of monohormonal cells from pluripotent stem cells comprising the following steps exposing pluripotent stem cells sequentially to:

a) a basal medium supplemented with at least CHIR99021 and Activin A;

b) a basal medium supplemented with Activin A;

c) a basal medium supplemented with at least FGF7;

d) a basal medium supplemented with retinoic acid, LDN 193189, and SANT1;

e) a basal medium supplemented with components consisting of 2 μM retinoic acid, 200 nM LDN193189, 0.25 UM SANT1, 10 ng/ml EGF and 10 ng/ml FGF2; and

f) a basal differentiation medium,

wherein step a is carried out for 1 day, step b is carried out for 2 days, step c is carried out for 2 days, step d is carried out for 4 days, step e is carried out for 14 days and step f is carried out for 7 days; and

wherein at least 90% of the cells in the resulting population are monohormonal and express insulin, but not glucagon, somatostatin, or ghrelin and are Glis3+.

2. The method of claim 1 , wherein the cells after step e express NKX6.1 and NEUROD1 at levels higher than the cells after step d.

3. The method of claim 1 , wherein the enriched population after step f has all of the following features:

a) at least 95% of the cells express PDX1;

b) at least 95% of the cells NKX2.2;

c) at least 85% of the cells express PAX6;

d) at least 90% of the cells express ISL1;

e) at least 50% of the cells express NKK6.1; and

f) at least 60% of the cells express UCN3.

4. The method of claim 1 , wherein the pluripotent stem cells are human pluripotent stem cells.

Assignments (1)

Continuity (2)

References Cited (29)