Hepatocytes and Chondrocytes from Adherent Placental StemCells; And CD34+ ,CD45- Placental Stem Cell-Enriched Cell Populations

• US 20080206343A1
• Filed: 02/12/2008
• Published: 08/28/2008
• Est. Priority Date: 02/12/2007
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Celularity, Inc.
• Inventors: Hariri, Robert J., Ye, Qian, Wang, Jia-Lun, Edinger, James W., Labazzo, Kristen S., Pereira, Marian, Abramson, Sascha Dawn
• Granted Patent: US 10,494,607 B2
• Time in Patent Office: Days
• US Class Current: 424/489
• CPC Class Codes: A01K 2267/0337 Animal models for infectious diseases
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C12N 2500/30 Organic components metal chelators C12N2500/10; calcitonin C12N2500/14; transferrin C12N2500/24
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/62 DMSO
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• 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 2533/32 Polylysine, polyornithine
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/74 Alginate
• 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 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/067 Hepatocytes

Abstract

Provided herein are methods and compositions for the production of hepatocytes from placenta stem cells. Further provided herein is the use of such hepatocytes in the treatment of, and intervention in, for example, trauma, inflammation, and degenerative disorders of the liver. Also provided herein are compositions and methods relating to combinations of nanofibrous scaffolds and adherent placental stem cells and methods of using the same in cartilage repair. Finally, provided herein are compositions and methods relating to nonadherent, CD34⁺CD45⁻ stem cells from placenta.

Claims

1. A method of producing a hepatocyte, comprising contacting a CD10⁺, CD34⁻

, CD105⁺ and CD200⁺ placental stem cell with sodium butyrate under conditions and for a time sufficient for said stem cell to exhibit a characteristic of a hepatocyte.

2. The method of claim 1, wherein said characteristic is production of asialogylcoprotein receptor, alpha-1-antitrypsin, albumin, cytochrome P450 activity, or the increased production of cytokeratin 18 relative to an undifferentiated placental stem cell.

3. The method of claim 1, wherein said culturing comprises encapsulating said stem sell in alginate-poly-L-lysine.

4. A hepatocyte or hepatocytic cell produced by the method of claim 1.

5. A method of treating a subject having a disease, disorder or condition associated with liver inflammation, comprising introducing the hepatocyte or hepatocytic cell of claim 4 to said subject.

6. The method of claim 5, wherein said disease, disorder or condition is cirrhosis or viral infection.

7. A mouse comprising human placental stem cell-derived hepatocytes or hepatogenic cells, wherein said mouse is produced by a method comprising the steps of:

a. irradiating said mouse with gamma radiation sufficient to kill substantially all of the endogenous bone marrow cells;

b. administering to said mouse sufficient bone marrow or bone marrow-derived cells from a NOD/SCID mouse to reconstitute the hematopoietic system of the mouse; and

c. transplanting to said mouse a plurality of hepatocytes or hepatogenic cells, wherein said hepatocytes or hepatogenic cells are differentiated from a plurality of CD10⁺, CD34⁻

, CD105⁺, CD200⁺ placental stem cells.

8. The mouse of claim 7, wherein said placental stem cell is additionally cytokeratin 18⁺.

9. The mouse of claim 7, wherein said hepatocytes or hepatogenic cells are administered into an ear pinna of the mouse.

10. The mouse of claim 8, wherein said hepatocytes or hepatogenic cells are infected with a virus.

11. The mouse of claim 10, wherein said virus is hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, or hepatitis E virus.

36. The cell population of claim 8, wherein said stem cell that is not CD34⁺ and CD45⁻

is contained within cord blood or placental blood.

12. A method of identifying an antiviral agent, comprising contacting the mouse of claim 12 with a compound of interest, wherein serum from said mouse has detectable levels of virus, and wherein said compound is an antiviral agent if said contacting results in a detectable reduction in the amount of said virus in serum from said mouse, compared to serum from said mouse not contacted with the compound of interest.

13. The method of claim 12, wherein said virus is hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, or hepatitis E virus.

14. The method of claim 13, wherein an antigen of said virus is detected.

15. The method of claim 13, wherein a nucleic acid of said virus is detected.

16. The method of claim 14, wherein said virus is hepatitis B virus.

17. The method of claim 14, wherein said antigen is HBeAg or HBsAg.

18. A composition comprising a plurality of cells encapsulated in alginate, wherein said cells are differentiated from placental stem cells, and wherein said cells express at least one marker of a hepatocyte not expressed by, or expressed to a detectably different degree than, an adherent placental stem cell that is CD10⁺, CD34⁻

, CD105⁺ and CD200⁺.

19. The composition of claim 18, wherein said alginate is in the form of beads.

20. The composition of claim 19, wherein said beads are from about 200 μ

m to about 800 μ

m in size.

21. The composition of claim 19, wherein said beads average about 500 μ

m in size.

22. A composition comprising isolated adherent CD10⁺, CD34⁻

, CD105⁺, CD200⁺ placental stem cells and an electrospun nanofibrous scaffold.

23. The composition of claim 22, wherein said nanofibrous scaffold comprises fibers of poly(L-lactic acid) (PLLA), poly lactic glycolic acid (PLGA), type I collagen, a copolymer of vinylidene fluoride and trifluoroethylene (PVDF-TrFE), poly(-caprolactone), poly(L-lactide-co-ε

-caprolactone) [P(LLA-CL)] (e.g., 75;

25), and/or a copolymer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and type I collagen.

24. The composition of claim 22, wherein said nanofibrous scaffold comprises fibers that average between about 250 nanometers and about 10 μ

m in thickness.

25. The composition of claim 22, wherein said composition is contacted with conditions in which the placental stem cells differentiate into chondrogenic cells or chondrocytes.

28. The cell population of claim 22, wherein at least 50% of cells in said population are CD34⁺ and CD45⁻

.

29. The cell population of claim 22, wherein at least 70% of cells in said population are CD34⁺ and CD45⁻

.

26. A method of making a composition comprising contacting adherent CD10⁺, CD34⁻

, CD105⁺, CD200⁺ placental stem cells with an electrospun nanofibrous scaffold, wherein said nanofibrous scaffold is made by electrospinning PLLA or PLGA at about 20 kV at about 30 cm needle to collector distance and about 0.05 mL/min. to about 0.1 mL/min flow rate, wherein said PLLA or PLGA are in solution at about 10% w/w to about 20% w/w.

27. An isolated cell population enriched for CD34⁺, CD45⁻

placental stem cells.

30. The cell population of claim 27, wherein at least 90% of cells in said population are CD34⁺ and CD45⁻

.

31. The cell population of claim 27, wherein said population comprises a stem cell that is not CD34⁺ and CD45⁻

.

32. The cell population of claim 31, wherein said stem cell that is not CD34⁺ and CD45⁻

is a CD34⁻

adherent placental stem cell.

33. The cell population of claim 32, wherein said adherent placental stem cell is CD200⁺, CD105⁺, CD90⁺, CD10⁺, CD34⁻

and CD45⁻

.

34. The cell population of claim 31, wherein said stem cell that is not CD34⁺ and CD45⁻

is a bone marrow-derived mesenchymal stem cell.

35. The cell population of claim 31, wherein said stem cell that is not CD34⁺ and CD45⁻

is a CD34⁺, CD45⁺ hematopoietic stem cell.

37. A method of producing a CD34⁺, CD45⁻

placental stem cell population, comprising selecting CD34⁺ cells from a population of placental cells to form a population of CD34⁺ placental cells, and removing from said population of CD34⁺ placental cells CD45⁺ cells, wherein a CD34⁺, CD45⁻

placental stem cell population is produced.