Re-aggregation of stem cell-derived pancreatic beta cells
AssigneeVertex Pharmaceuticals Incorporated
InventorsYeh-Chuin POH, George HARB, Felicia J. PAGLIUCA
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Abstract
The present application discloses cell clusters resembling the function and characteristics of endogenous pancreatic islets, and methods for making and using such cell clusters.
Claims28
1-24. (canceled).
A method of treating or preventing a disease in a human subject in need thereof, the method comprising administering a cell cluster to the subject, the cell cluster comprising at least one non-native pancreatic β-cell that exhibits an in vitro glucose-stimulated insulin secretion response when exposed to a glucose challenge, wherein the cell cluster is an unsorted cell cluster, and wherein the cell cluster comprises:
- (a) at least about 35% cells that express NKX6.1 and C-peptide;
- (b) at least about 95% cells that express chromogranin A;
- and (c) at most about 2% cells that express SOX9.
A method of producing an in vitro cell cluster, the method comprising:
- (a) obtaining a first cell cluster comprising at least one NKX6.1-positive and C-peptide-positive cell;
- (b) dissociating a plurality of cells from the first cell cluster, wherein the dissociating does not comprise an active cell sorting process to form a dissociated plurality of cells;
- (c) cryopreserving the dissociated plurality of cells;
- (d) thawing the cryopreserved dissociated plurality of cells;
- and
- (e) culturing the thawed dissociated plurality of cells from (d) in a serum-free culture medium and allowing at least a portion of the plurality of cells to form a second in vitro cell cluster, wherein:
- (i) the second in vitro cell cluster comprises a higher percentage of cells that express chromogranin A as compared to the first cell cluster;
- (ii) the second in vitro cell cluster comprises a higher percentage of cells that express NKX6.1 and C-peptide as compared the first cell cluster;
- (iii) the second in vitro cell cluster comprises a lower percentage of cells that express SOX2 as compared the first cell cluster;
- (iv) the second in vitro cell cluster comprises a lower percentage of cells that express SOX9 as compared the first cell cluster;
- and
- (v) wherein the second in vitro cell cluster comprises at least one non-native pancreatic β cell that exhibits an in vitro glucose-stimulated insulin secretion response when exposed to a glucose challenge.
Dependencies are inferred from the claim text.