Regeneration of pancreatic islets by amniotic fluid stem cell therapy

• US 9,056,093 B2
• Filed: 01/06/2006
• Issued: 06/16/2015
• 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, Milanesi, Anna, Soker, Shay
• Primary Examiner(s): Singh, Anoop
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US11/327,074
• Publication Number:
  US 20070031384A1
• Time in Patent Office: 3,448 Days
• Field of Search: 435/325, 435/366
• US Class Current: 1/1
• 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 insulin-dependent diabetes in a human subject in need thereof, comprising the steps of:

(i) providing a pharmaceutical composition comprising human amniotic fluid-derived stem cells, wherein said amniotic fluid-derived stem cells;

(a) specifically bind c-kit antibodies at the time of collection,

(b) are transfected with a nucleic acid expressing Pdx-1,

(c) express the cell surface marker CD44 and do not express CD34 and CD105 cell surface markers, and

(ii) administering the pharmaceutical composition of step (i) to said human subject in an amount effective to treat said insulin-dependent diabetes.

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

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

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

(c) 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 cells are not differentiated into pancreatic-like cells prior to said administering step.

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

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

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

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

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

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

13. A pharmaceutical composition for the treatment of diabetes comprising amniotic fluid-derived stem cells, wherein said amniotic fluid-derived stem cells:

(i) specifically bind c-kit antibodies at the time of collection,

(ii) are transfected with a nucleic acid expressing Pdx-1,

(iii) express the cell surface marker CD44 and do not express CD34 and CD105 cell surface markers, and

(iv) are provided in a treatment-effective amount.

14. The composition of claim 13, wherein said composition comprises an aqueous carrier.

15. The composition of claim 13 in unit dosage form.

16. The composition of claim 13, wherein said composition is an injectable composition.

17. The composition of claim 13, wherein said amniotic fluid-derived stem cells are encapsulated in an insulin-permeable capsule.

18. The composition of claim 13, wherein said amniotic fluid-derived stem cells are unencapsulated.

19. The composition of claim 13, wherein said amniotic fluid-derived stem cells are human cells.

20. The composition of claim 13, wherein said amniotic fluid-derived stem cells:

(i) are collected from amniotic fluid of a mammalian donor,

(ii) differentiate upon appropriate stimulation into at least osteogenic, adipogenic, myogenic, neurogenic, hematopoietic, 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, and

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

21. The composition of claim 13, wherein said amniotic fluid-derived stem cells express the transcription factor Oct-4, SSEA-1, Sca-1, and CD90.

22. The composition of claim 13, wherein said nucleic acid comprises DNA or RNA.

23. The composition of claim 13, wherein said nucleic acid is RNA.

24. The composition of claim 13, wherein said composition is isotonic with human blood.

25. The composition of claim 13, wherein said composition comprises from about 1×

10⁵ to about 1×

10⁹ of said cells.

26. The composition of claim 14, wherein said cells are provided as a suspension in the carrier.

27. A pharmaceutical composition for the treatment of diabetes comprising amniotic fluid-derived stem cells transfected with a nucleic acid expressing Pdx-1, wherein said amniotic fluid-derived stem cells:

(i) are collected from amniotic fluid of a mammalian donor,

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

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

(iv) do not form teratomas when grown in vivo,

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

(vi) express the cell surface marker CD44 and do not express CD34 and CD 105 cell surface markers, and

(vii) are provided in a treatment-effective amount,

wherein said composition is an injectable composition that is isotonic with human blood.

28. The composition of claim 27, wherein said amniotic fluid-derived stem cells are human cells.

29. The composition of claim 27, wherein said nucleic acid comprises DNA or RNA.

30. The composition of claim 27, wherein said nucleic acid is RNA.

31. The composition of claim 27, wherein said composition comprises from about 1×

10⁵ to about 1×

10⁹ of said cells.

32. The composition of claim 27, wherein said cells are provided as a suspension in a carrier.