Placental stem cell populations

• US 8,202,703 B2
• Filed: 07/30/2010
• Issued: 06/19/2012
• Est. Priority Date: 12/29/2005
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Edinger, James, Ye, Qian, Wang, Jai-Lun, Hariri, Robert J.
• Primary Examiner(s): Long, Scott
• Application Number:
  US12/848,007
• Publication Number:
  US 20100297689A1
• Time in Patent Office: 690 Days
• Field of Search: None
• US Class Current: 435/29
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 35/644 Beeswax; Propolis; Royal jelly; Honey
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• 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

Abstract

The present invention provides placental stem cells and placental stem cell populations, and methods of culturing, proliferating and expanding the same. The invention also provides methods of differentiating the placental stem cells. The invention further provides methods of using the placental stem cells in assays and for transplanting.

Claims

1. A population of isolated adherent placental stem cells, wherein said stem cells:

express genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells (BM-MSCs);

wherein said genes comprise ELOVL2, ST3GAL6, ST6GALNAC5, or SLC12A8;

wherein at least 70% of said placental stem cells are non-maternal in origin; and

wherein said cells have been passaged at least three times and cultured under growth conditions.

2. The population of claim 1, wherein at least 90% of said placental stem cells are non-maternal in origin.

3. The population of claim 1 present in a form suitable for intravenous administration.

4. The population of claim 1, wherein said cells have the capacity to differentiate into cells having characteristics of neural cells.

5. The population of claim 1, wherein said genes further comprise CD200 and NUAK1.

6. The population of claim 1, wherein said genes further comprise ARTS-1, IER3, IL6, KRT18, LRAP, MEST, NFE2L3, or TGFB2.

7. The population of claim 1, wherein said expression of said genes is detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells over 3 population doublings.

8. The population of claim 1, wherein said expression of said genes is detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells over 11-14 population doublings.

9. The population of claim 1, wherein said expression of said genes is detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells over 24-38 population doublings.

10. The population of claim 1, wherein said population comprises at least 1 ×

10⁹ cells.

11. The population of claim 1, wherein said population has undergone at least 5 population doublings.

12. The population of claim 1, additionally comprising stem cells that are not obtained from placental tissue.

13. The population of claim 12, wherein said stem cell not obtained from a placenta is a mesenchymal stem cell.

14. The population of claim 12, wherein said stem cells not obtained from a placenta are hematopoietic progenitor cells.

15. The population of claim 1, wherein said cells have been cryopreserved.

16. The population of claim 1, wherein said cells express said one or more genes at a detectably higher level than an equivalent number of BM-MSCs in medium comprising DMEM-LG and MCDB-201;

2% fetal calf serum, 1×

insulin-transferrin-selenium, 1×

lenolenic-acid-bovine-serum-albumin, 10<sup>−

9</sup> M dexamethasone, 10<sup>−

4</sup> M ascorbic acid 2-phosphate, 10 ng/ml epidermal growth factor, and 10 ng/ml platelet derived-growth factor.

17. The population of claim 1, wherein said placental cells have the ability to replicate 10-40 times in culture.

18. The population of claim 1, wherein said placental cells have the ability to replicate 10-40 times in culture in DMEM and fetal bovine serum.

19. The population of claim 1, wherein said placental cells are isolated from a human postpartum placenta by digestion using trypsin, or are cultured from cells isolated from a human postpartum placenta by digestion using trypsin.

21. The population of claim 1, wherein said population comprises 1×

10⁸ to 5×

10⁸ placental stem cells.

22. The population of claim 1, wherein said population comprises 5×

10⁸ to 1×

10⁹ placental stem cells.

23. The population of claim 1, wherein said population comprises 1×

10⁹ to 5×

10⁹ placental stem cells.

24. The population of claim 1, wherein said placental stem cells have been passaged 4 to 8 times.

25. The population of claim 1, wherein said placental stem cells have been passaged 6 times.

26. The population of claim 1, wherein said placental cells have undergone 18 to 26 population doublings.

27. The population of claim 1, wherein said placental stem cells have a doubling time of three to five days.

28. The population of claim 1, wherein said population is present in a composition comprising dimethylsulfoxide (DMSO), human serum albumin (HSA), or dextran.

29. The population of claim 1, wherein said placental stem cells differentiate into cells having a characteristic of chondrogenic cells when cultured in DMEM comprising 15% cord blood serum and 0.01 μ

g/mL transforming growth factor beta (TGFβ

); and

wherein said characteristic of chondrogenic cells is positive staining with Alcian Blue stain.

30. The population of claim 1, wherein said placental stem cells differentiate into cells having a characteristic of osteogenic cells when cultured in DMEM comprising 15% cord blood serum, 0.1 μ

M dexamethasone, 0.05 mM ascorbic acid-2-phosphate, and 10 mM beta glycerophosphate; and

wherein said characteristic of osteogenic cells is staining with von Kossa stain or production of mRNA for alkaline phosphatase as determined by RT-PCR.

31. The population of claim 1, wherein at least 90% of cells in the population are said placental stem cells.

32. The population of claim 1, wherein at least 95% of cells in the population are said placental stem cells.

33. The population of claim 1, wherein at least 99% of cells in the population are said placental stem cells.

34. The population of claim 1, wherein said cells express said genes at least at a three-fold higher level than an equivalent number of BM-MSCs.

35. The population of claim 1, wherein said genes comprise ELOVL2.

36. The population of claim 1, wherein said genes comprise ST3GAL6.

37. The population of claim 1, wherein said genes comprise ST6GALNAC5.

38. The population of claim 1, wherein said genes comprise SLC12A8.

20. The population of clam 1, wherein said placental cells have been separated from at least 90% of cells from the placenta from which the placental cells are isolated.