Placental stem cells and uses thereof

• US 20040161419A1
• Filed: 10/22/2003
• Published: 08/19/2004
• Est. Priority Date: 04/19/2002
• Status: Abandoned Application

• Current Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Original Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Inventors: Miki, Toshio, Strom, Stephen C.
• Application Number:
  US10/691,468
• Publication Number:
  US 20040161419A1
• Time in Patent Office: Days
• US Class Current: 424/93.21
• 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: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• 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 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/148 Transforming growth factor alpha [TGF-a]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• 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 2501/39 Steroid hormones
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic 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 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/069 : Vascular Endothelial cells

Abstract

The present invention features novel placental stem cells and provides methods and compositions for the therapeutic uses of placental stem cells or placental stem cells that have been induced to differentiate into a desired tissue type into a recipient host in amounts sufficient to result in production of the desired cell type, e.g, hepatocytes, neural cells, pancreatic cells, vascular endothelial cells, cardiomyocytes.

Claims

1. A composition comprising a placental stem cell which expresses at least one marker selected from the group consisting of:

Thy-1, OCT-4, SOX2, SSEA3, SSEA4, TRA1-60, TRA1-81, Lefty A, FGF-4, Rex-1 and TDGF-1.

3. A composition of claim 1 that is ATCC deposit No. ______.

4. A pharmaceutical composition comprising an effective amount of the composition of claim 1 and a pharmaceutically acceptable carrier.

5. A method of making a cardiomyocyte comprising culturing a stem cell of claim 1 in a media that contains an appropriate amount of ascorbic acid 2-phosphate under appropriate conditions and for a sufficient period of time for the stem cell to differentiate into a cardiomyocyte.

6. A cardiomyocyte obtained from the process of claim 5, which expresses at least one marker selected from the group consisting of:

MLC-2A, MLC-2V, hANP, cTnT, alpha-actinin, GATA-4 and Nkx 2.5.

7. A cardiomyocyte obtained from the process of claim 5, which expresses at least two markers selected from the group consisting of:

MLC-2A, MLC-2V, hANP, cTnT, alpha-actinin, GATA-4 and Nkx 2.5.

9. A pharmaceutical composition comprising an effective amount of a cardiomyocyte of claim 6 and a pharmaceutically acceptable carrier.

10. A method of determining whether a test agent is toxic to a cardiomyocyte, comprising contacting the cardiomyocyte of claim 6 with an appropriate amount of the test agent for a time sufficient for a toxic effect on the cardiomyocyte to be detected, and determining whether the test agent has a toxic effect on the cardiomyocyte.

11. A method of determining a metabolic product of a test agent comprising contacting the cardiomyocyte of claim 6 with an appropriate amount of the test agent for a time sufficient for the test agent to be metabolized, and detecting the presence of the metabolized product.

12. A method of making a hepatocyte comprising culturing a stem cell of claim 1 in a media that contains an appropriate amount of dexamethasone, ITS, EGF, FGF-2, FGF-4, FGF-7, HGF, phenobarbital, Type-I collagen or a combination thereof under appropriate conditions and for a sufficient period of time for the stem cell to differentiate into a hepatocyte.

13. A hepatocyte obtained from the process of claim 12, which expresses at least one marker selected from the group consisting of:

albumin, CYP1A1, CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2D6, CYP3A4, AFP, A1AT, HNF1, HNF4 and C/EBP alpha.

14. A hepatocyte obtained from the process of claim 12, which expresses at least two markers selected from the group consisting of:

albumin, CYP1A1, CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2D6, CYP3A4, AFP, A1AT, HNF1, HNF4 and C/EBP alpha.

16. A pharmaceutical composition comprising an effective amount of a hepatocyte of claim 13 and a pharmaceutically acceptable carrier.

17. A method of determining whether a test agent is toxic to a hepatocyte comprising contacting the hepatocyte of claim 13 with an appropriate amount of the test agent for a time sufficient for a toxic effect on the hepatocyte to be detected, and determining whether the test agent has a toxic effect on the hepatocyte.

18. A method of determining a metabolic product of a test agent comprising contacting the hepatocyte of claim 13 with an appropriate amount of the test agent for a time sufficient for the test agent to be metabolized, and detecting the presence of the metabolized product.

19. A method of making a pancreatic cell comprising culturing a stem cell of claim 1 in a media that contains an appropriate amount of nicotinamide, dexamethasone, ITS, matrigel or a combination thereof under appropriate conditions and for a sufficient period of time for the stem cell to differentiate into a pancreatic cell.

20. A pancreatic cell obtained from the process of claim 19, which expresses at least one marker selected from the group consisting of:

Pax6, Pdx1, insulin, glucagon, and Nkx2.2.

21. A pancreatic cell obtained from the process of claim 19, which expresses at least two markers selected from the group consisting of:

Pax6, Pdx1, insulin, glucagon, and Nkx2.2.

23. A pharmaceutical composition comprising an effective amount of a pancreatic cell of claim 20 and a pharmaceutically acceptable carrier.

24. A method of determining whether a test agent is toxic to a pancreatic cell comprising contacting the pancreatic cell of claim 20 with an appropriate amount of the test agent for a time sufficient for a toxic effect on the pancreatic cell to be detected, and determining whether the test agent has a toxic effect on the pancreatic cell.

25. A method of determining a metabolic product of a test agent comprising contacting the pancreatic cell of claim 20 with an appropriate amount of the test agent for a time sufficient for the test agent to be metabolized, and detecting the presence of the metabolized product.

26. A method of making a neural cell comprising culturing a stem cell of claim 1 in a media that contains an appropriate amount of trans-retinoic acid or FGF-4 under appropriate conditions and for a sufficient period of time for the stem cell to differentiate into a neural cell.

27. A neural cell obtained from the process of claim 26, which expresses at least one marker selected from the group consisting of:

GFAP, CNP, beta-tubulin III, Nestin, GAD, NSE, NF-M and MBP.

28. A neural cell obtained from the process of claim 26, which expresses at least two markers selected from the group consisting of:

GFAP, CNP, beta-tubulin III, Nestin, GAD, NSE, NF-M and MBP.

30. A pharmaceutical composition comprising an effective amount of a neural cell of claim 27 and a pharmaceutically acceptable carrier.

31. A method of determining whether a test agent is toxic to a neural cell comprising contacting the neural cell of claim 27 with an appropriate amount of the test agent for a time sufficient for a toxic effect on the neural cell to be detected, and determining whether the test agent has a toxic effect on the neural cell.

32. A method of determining a metabolic product of a test agent comprising contacting the neural cell of claim 27 with an appropriate amount of the test agent for a time sufficient for the test agent to be metabolized, and detecting the presence of the metabolized product.

33. A method of making a vascular endothelial cell comprising culturing a stem cell of claim 1 in a media that contains matrigel under appropriate conditions and for a sufficient period of time for the stem cell to differentiate into a vascular endothelial cell.

34. A vascular endothelial cell obtained from the process of claim 33, which expresses the FLT-1 marker.

35. A vascular endothelial cell obtained from the process of claim 33, which has physical characteristics of cells shown in FIG. 10.

37. A pharmaceutical composition comprising an effective amount of a vascular endothelial cell of claim 34 and a pharmaceutically acceptable carrier.

38. A method of determining whether a test agent is toxic to a vascular endothelial cell comprising contacting the vascular endothelial cell of claim 34 with an appropriate amount of the test agent for a time sufficient for a toxic effect on the vascular endothelial cell to be detected, and determining whether the test agent has a toxic effect on the vascular endothelial cell.

39. A method of determining a metabolic product of a test agent comprising contacting the vascular endothelial cell of claim 34 with an appropriate amount of the test agent for a time sufficient for the test agent to be metabolized, and detecting the presence of the metabolized product.

2. A composition comprising a placental stem cell which expresses at least two markers selected from the group consisting of:

Thy-1, OCT-4, SOX2, SSEA3, SSEA4, TRA1-60, TRA1-81, Lefty A, FGF-4, Rex-1 and TDGF-1.

8. A cardiomyocyte that is ATCC deposit No. ______.

15. A hepatocyte that is ATCC deposit No. ______.

22. A pancreatic cell that is ATCC deposit No. ______.

29. A neural cell that is ATCC deposit No. ______.

36. A vascular endothelial cell that is ATCC deposit No. ______.