Regeneration of pancreatic islets by amniotic fluid stem cell therapy

• US 20070031384A1
• Filed: 01/06/2006
• Published: 02/08/2007
• Est. Priority Date: 01/07/2005
• Status: Active Grant

• Current Assignee: Wake Forest University Health Sciences
• Original Assignee: Wake Forest University Health Sciences
• Inventors: Atala, Anthony, Soker, Shay, Milanesi, Anna
• Granted Patent: US 9,056,093 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/03 from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

A method of treating diabetes in a mammalian subject is carried out by: (a) providing mammalian amniotic fluid stem cells, and then (b) administering the cells to the subject in an amount effective to treat diabetes. Optionally, the cells may be differentiated into pancreatic-like cells or at least treated to initiate subsequent differentiation into-pancreatic-like cells, prior to administration.

Claims

1. A method of treating diabetes in a mammalian subject in need thereof, comprising the steps of:

(a) providing mammalian amniotic fluid stem cells, wherein said cells are optionally differentiated into pancreatic-like cells; and

then

(b) administering said cells to said subject in an amount effective to treat diabetes.

2. The method of claim 1, wherein said amniotic fluid stem cells:

(i) are collected from a mammalian donor,

(ii) differentiate upon appropriate stimulation into at least osteogenic, adipogenic, myogenic, neurogenic, hematopoitic, and endothelial cell lines,

(iii) proliferate through at least 200 population doublings when grown in vitro,

(iv) do not require feeder cell layers to grow in vitro,

(v) specifically bind c-kit antibodies at the time of collection; and

(vi) do not form teratomas when grown in vivo.

3. The method of claim 1, wherein said administering step is carried out by intraveneous or intraarterial injection.

4. The method of claim 1, wherein said cells are administered in an amount of from about 1×

10⁵ to about 1×

10⁹ cells per subject.

5. The method of claim 1, wherein said subject is effectively treated for said diabetes for at least two months after said administering step.

6. The method of claim 1, wherein said diabetes is insulin-dependent diabetes.

7. The method of claim 1, wherein said cells are not differentiated into pancreatic-like cells prior to said administering step.

8. The method of claim 1, wherein said cells are not differentiated into pancreatic-like cells prior to said administering step, and wherein said cells are administered into the pancreas of said subject.

9. The method of claim 1, wherein said cells are differentiated into pancreatic-like cells prior to said administering step.

10. The method of claim 1, wherein said cells are differentiated into pancreatic-like cells prior to said administering step by culturing said cells in a medium containing a pancreatic differentiation factor.

11. The method of claim 1, wherein said cells are differentiated into pancreatic-like cells prior to said administering step by expressing a recombinant or endogeneous pancreatic differentiation factor therein.

12. The method of claim 11, wherein said differenting step is carried out by transducing said cells with a vector that contains a nucleic acid encoding a pancreatic differentiation factor and expresses said differentiation factor in said cells.

13. The method of claim 12, wherein said vector is a plasmid, cosmid, bacteriophage, DNA virus, RNA virus, or retrovirus vector.

14. The method of claim 12, wherein said pancreatic differentiation factor is Pdx1, Ngn3, Nkx6.1, Nkx2.1, Pax6, or Pax4.

15. The method of claim 12, wherein said pancreatic differentiation factor is transiently expressed in said cells.

16. The method of claim 1, wherein said cells are allogenic with respect to said subject.

17. The method of claim 1, wherein said cells are xenogenic with respect to said subject.

18. The method of claim 1, wherein said cells are encapsulated in an insulin-permeable capsule.

19. The method of claim 1, wherein said cells are unencapsulated.

20. The method of claim 1, wherein said cells are human cells.

21. The method of claim 1, wherein said subject is a human subject.

22. The method of claim 1, wherein said subject is an adult subject.

23. The method of claim 1, wherein said subject is an adolescent subject.

24. The method of claim 1, wherein said subject is a juvenile subject.

25. The method of claim 1, wherein said providing step (a) is carried out by (i) collecting amniotic fluid stem cells from a mammalian donor, and then (ii) expanding said amniotic fluid stem cells in vitro.

34. The method of claim 1, wherein said amniotic fluid stem cells express the transcription factor Oct-4 (Pou5f1), SSEA-1 (Stage Specific Embryonic Antigen 1), Sca-1 (Ly-6A/E), CD90 (Thy-1), and CD44 (Hyaluronate receptor. Ly-24, Pgp-1).

26. A pharmaceutical composition useful for the treatment of diabetes comprising, pancreatic-like cells in a pharmaceutically acceptable carrier, wherein said pancreatic-like cells are differentiated from amniotic fluid stem cells.

27. The composition of claim 26, wherein said carrier is an aqueous carrier.

28. The composition of claim 26 in unit dosage form.

29. The composition of claim 26, wherein said composition is an injectable composition.

30. The composition of claim 26, wherein said cells are encapsulated in an insulin-permeable capsule.

31. The composition of claim 26, wherein said cells are unencapsulated.

32. The composition of claim 26, wherein said cells are human cells.

33. The composition of claim 26, wherein said amniotic fluid stem cells:

(i) are collected from a mammalian donor,

(ii) differentiate upon appropriate stimulation into at least osteogenic, adipogenic, myogenic, neurogenic, hematopoitic, and endothelial cell lines,

(iii) proliferate through at least 200 population doublings when grown in vitro,

(iv) do not require feeder cell layers to grow in vitro,

(v) specifically bind c-kit antibodies at the time of collection; and

(vi) do not form teratomas when grown in vivo.

35. The composition of claim 26, wherein said amniotic fluid stem cells express the transcription factor Oct-4 (Pou5f1), SSEA-1 (Stage Specific Embryonic Antigen 1), Sca-1 (Ly-6A/E), CD90 (Thy-1), and CD44 (Hyaluronate receptor. Ly-24, Pgp-1).