ENCAPSULATION OF PANCREATIC CELLS DERIVED FROM HUMAN PLURIPOTENT STEM CELLS

• US 20100124564A1
• Filed: 11/13/2009
• Published: 05/20/2010
• Est. Priority Date: 11/14/2008
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Martinson, Laura, Kroon, Evert, Kelly, Olivia, Agulnick, Alan, Green, Chad, Baetge, Emmanuel E.
• Granted Patent: US 8,278,106 B2
• Time in Patent Office: Days
• US Class Current: 424/424
• CPC Class Codes: A01N 1/0221 Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
• CPC Class Codes: A61F 2/022 Artificial gland structures using bioreactors hemodialysis A61M1/16; hemofiltration A61M1/34; other extracorporeal blood treatment A61M1/36
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2300/62 Encapsulated active agents, e.g. emulsified droplets
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 27/14 Macromolecular materials
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61M 2202/07 Proteins
• CPC Class Codes: A61M 2205/04 implanted
• CPC Class Codes: A61M 31/00 Devices for introducing or retaining media, e.g. remedies, in cavities of the body A61M25/00 takes precedence ; introducing or retaining ophthalmic products into the ocular cavities A61F9/0008
• CPC Class Codes: A61M 31/002 Devices for releasing a drug at a continuous and controlled rate for a prolonged period of time artificial gland structures or devices A61F2/022; intra-uterine contraceptive devices A61F6/14; tampons for introducing into the vagina A61F13/20, A61L15/00; suppositories or bougies for intra-vaginal or intra-uterine application A61K9/02; physical forms of medicinal preparations for sustained or differential drug release A61K9/20, A61K9/50
• CPC Class Codes: A61M 5/00 Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests vaccination appliances for veterinary use A61D1/025 ; tube connectors, tube couplings, valves or branch units specially adapted for medical use A61M39/00; containers specially adapted for medical or pharmaceutical purposes A61J1/00; combinations of vial and syringe for mixing or transferring their contents A61J1/20; holders for containers for collecting, storing or administering blood or medical fluids A61J1/16
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C12N 2501/115 : Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 : Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/119 : Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/16 : Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/19 : Growth and differentiation factors [GDF]
• CPC Class Codes: C12N 2501/385 : of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/41 : Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/415 : Wnt; Frizzeled
• CPC Class Codes: C12N 2506/02 : from embryonic cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells

Abstract

The present invention relates to methods for encapsulating pancreatic progenitors in a biocompatible semi-permeable encapsulating device. The present invention also relates to production of human insulin in a mammal in response to glucose stimulation.

Claims

1. A method for producing insulin in vivo in a mammal, said method comprising:

(a) providing an in vitro human PDX1-positive pancreatic progenitor cell population into an implantable semi-permeable device;

(b) implanting the device with the cell progenitor cell population into a mammalian host; and

(c) maturing the progenitor cell population in said device in vivo such that the progenitor cell population comprises endocrine and acinar cells, wherein at least some of the endocrine cells are insulin secreting cells that produce insulin in response to glucose stimulation in vivo, thereby producing insulin in vivo to the mammal.

2. The method of claim 1, further comprising maturing PDX1-positive pancreatic progenitor cells to acinar or duct cells.

3. The method of claim 1, wherein the device is first prevascularized by implanting in the mammal.

4. The method of claim 1, wherein the device comprises a plurality of welds for maximizing surface area to volume ratio of the human PDX1-positive pancreatic progenitor cell population in the device.

5. The method of claim 1, wherein the device comprises a plurality of welds to increase vascularization by the host.

6. The method of claim 1, wherein the device is refillable.

7. The method of claim 1, wherein the device is expandable.

8. The method of claim 1, wherein the in vivo cell population is monitored.

9. The method of claim 1, wherein greater than 25 pM of insulin is detectable in the serum of the mammal post glucose-stimulation.

10. The method of claim 1, wherein greater than 50 pM of human C-peptide is detectable in the serum of the mammal post glucose-stimulation.

11. The method of claim 1, wherein the cell population is protected from one or more cells of the host mammal.

12. The method of claim 1, wherein the mammal is immuno-suppressed.

13. The method of claim 1, wherein providing a human PDX1-positive pancreatic progenitor cell population comprises thawing a cell population.

14. The method of claim 1, wherein providing a human PDX1-positive pancreatic progenitor cell population further comprises enriching the cell population for PDX1/NKX6.1 co-positive pancreatic progenitor cells.

15. The method of claim 1, wherein providing a human PDX1-positive pancreatic progenitor cell population further comprises enriching the cell population for PDX1/NKX6.1 co-positive pancreatic progenitor cell by contacting the cell population with an antibody that binds to CD142 antigen and enriching for said CD142 binding cells.

16. A cell encapsulating assembly for implanting into a mammalian host, said assembly comprising at least one chamber for encapsulating living cells, wherein the assembly comprises a first seal at a peripheral edge of the assembly, thereby forming the encapsulating assembly, and a second seal which effectively reduces the chamber volume but increases device surface area.

17. The assembly of claim 16, wherein the assembly comprises a semi-permeable membrane.

18. The assembly of claim 16, wherein the assembly has a third or fourth seal and further reduces the chamber volume.

19. The assembly of claim 16 further comprising of at least one loading port.

20. The assembly of claim 16 comprising two loading ports.

21. The assembly of claim 16, wherein the assembly further comprises living cells.

22. The assembly of claim 21, wherein the living cells are human PDX1-positive pancreatic progenitor cells.

34. The method of claim 33 wherein the cell encapsulation assembly is the assembly of claim 16.

23. A cryopreserved human pancreatic progenitor cell population, wherein the cell population is suitable for transplantation into a mammal.

24. The cell population of claim 23, wherein cells of the cell population are capable of maturing into islet or acinar cells in the mammal.

25. The cell population of claim 23, wherein cells of the cell population are capable of maturing into beta cells which are capable of secreting insulin in response to glucose stimulation.

26. The cell population of claim 23, wherein the pancreatic progenitor cell population comprises PDX1 positive pancreatic progenitor cells.

27. A method of preserving an in vitro population of cells suitable for transplantation, said method comprising:

a) contacting a population of human pancreatic progenitor cells with a cryopreservation solution; and

b) decreasing the temperature of the progenitor cells for cryopreservation to less than about 0°

C., thereby preserving an in vitro population of pancreatic progenitor cells suitable for transplantation.

28. The method of claim 27, wherein the temperature of the progenitor cells for cryopreservation is decreased to less than about −

50°

C.

29. The method of claim 27, wherein the temperature of the progenitor cells for cryopreservation is decreased to less than about −

100°

C.

30. The method of claim 27, wherein the temperature of the progenitor cells for cryopreservation is decreased to less than about −

150°

C.

31. The method of claim 27, wherein the temperature of the progenitor cells for cryopreservation is decreased to less than about −

190°

C.

32. A method of providing an in vitro population of cells suitable for transplantation, said method comprising increasing the temperature of a cryopreserved cell population to obtain a population of pancreatic progenitor cells suitable for transplantation.

33. The method of claim 32 further comprising the step of providing the cell population to a cell encapsulation assembly.