AMNION DERIVED ADHERENT CELLS

• US 20100124569A1
• Filed: 11/19/2009
• Published: 05/20/2010
• Est. Priority Date: 11/19/2008
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Labazzo, Kristen, Paredes, Jennifer, Jankovic, Vladimir, Law, Eric, Padliya, Neerav D., Francki, Aleksandar, Kaplunovsky, Aleksandr, Wang, Jia-Lun, Edinger, James W., ABBOT, Stewart
• Granted Patent: US 8,367,409 B2
• Time in Patent Office: Days
• US Class Current: 424/484
• 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 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly

Abstract

Provided herein are novel angiogenic cells from amnion, referred to as amnion derived adherent cells, and populations of, and compositions comprising, such cells. Further provided herein are methods of obtaining such cells and methods of using the cells in the treatment of individuals.

Claims

1. An isolated amnion derived adherent cell, wherein said cell is adherent to tissue culture plastic, and wherein said cell is OCT-4⁻

(octamer binding protein

     4) as determined by RT-PCR.

2. The isolated cell of claim 1, wherein said cell is HLA-G⁻

, as determined by RT-PCR.

3. The isolated cell of claim 1, wherein said cell is additionally CD49f⁺, as determined by flow cytometry.

4. The isolated cell of claim 3, wherein said cell is OCT-4⁻

, HLA-G⁻

and CD49f⁺.

5. The isolated cell of claim 1, wherein said cell is CD90⁺, CD105⁺, or CD117⁻

as determined by flow cytometry.

6. The isolated cell of claim 5, wherein said cell is CD90⁺, CD105⁺, and CD117⁻

.

7. The isolated cell of claim 6, wherein said cell is OCT-4⁻

and HLA-G⁻

, as determined by RT-PCR, and CD49f⁺, CD90⁺, CD105⁺, and CD117⁻

as determined by flow cytometry.

8. The isolated cell of claim 1, wherein said cell is VEGFR1/Flt-1⁺ (vascular endothelial growth factor receptor 1) and VEGFR2/KDR⁺ (vascular endothelial growth factor receptor 2), as determined by immunolocalization

9. The isolated cell of claim 1, wherein said cell is one or more of CD9⁺, CD10⁺, CD44⁺, CD54⁺, CD98⁺, Tie-2⁺ (angiopoietin receptor), TEM-7⁺ (tumor endothelial marker 7), CD31⁻

, CD34⁻

, CD45⁻

, CD133⁻

, CD143⁻

(angiotensin-I-converting enzyme, ACE), CD146⁻

(melanoma cell adhesion molecule), or CXCR4⁻

(chemokine (C-X-C motif) receptor

     4) as determined by immunolocalization.

10. The isolated cell of claim 1, wherein said cell is CD9⁺’

CD10⁺, CD44⁺, CD54⁺, CD98⁺, Tie-2⁺ (angiopoietin receptor), TEM-7⁺ (tumor endothelial marker

     7), CD31⁻

, CD34⁻

, CD45⁻

, CD133⁻

, CD143⁻

, CD146⁻

, and CXCR4⁻

as determined by immunolocalization.

11. The isolated cell of claim 1, wherein said cell is VE-cadherin⁻

as determined by immunolocalization.

12. The isolated cell of claim 1, wherein said cell is additionally positive for CD105⁺ and CD200⁺ as determined by immunolocalization.

13. The isolated cell of claim 1, wherein said cell does not express CD34 as detected by immunolocalization after exposure to 50 ng/mL VEGF for 7 days.

14. An isolated population of cells comprising the cell of claim 1.

15. The isolated population of cells of claim 14, wherein at least 50% of the cells in said population are the cells of claim 1.

16. The isolated population of cells of claim 14, wherein at least 90% of the cells in said population are the cells of claim 1.

17. An isolated population of cells comprising the cell of claim 7.

18. The isolated population of cells of claim 17, wherein at least 50% of the cells in said population are the cells of claim 7.

19. The isolated population of cells of claim 11, wherein at least 90% of the cells in said population are the cells of claim 7.

20. An isolated population of cells comprising the cell of claim 4, wherein said isolated population of cells is not an amnion.

21. The isolated population of cells of claim 14, wherein at least 50% of the cells in said population are the cells of claim 4.

22. The isolated population of cells of claim 14, wherein at least 90% of the cells in said population are the cells of claim 4.

23. A composition comprising the isolated amnion derived adherent cell of claim 1 or claim 7.

24. The isolated population of cells of claim 14, wherein said second type of cell is an embryonic stem cell, a blood cell, a stem cell isolated from peripheral blood, a stem cell isolated from placental blood, a stem cell isolated from placental perfusate, a stem cell isolated from placental tissue, a stem cell isolated from umbilical cord blood, an umbilical cord stem cell, a bone marrow-derived mesenchymal stem cell, a bone marrow-derived mesenchymal stromal cell, a hematopoietic stem cell, a somatic stem cell, a chondrocyte, a fibroblast, a muscle cells, an endothelial cell, an angioblast, an endothelial progenitor cell, a pericyte, a cardiomyocyte, a myocyte, a cardiomyoblast, a myoblast, an embryonic stem cell, or a cell manipulated to resemble an embryonic stem cell.

25. The isolated population of cells of claim 24, wherein said second type of cell comprises at least 10% of cells in said population.

26. The isolated population of cells of claim 24, wherein said second type of cell comprises at least 25% of cells in said population.

27. The isolated population of cells of claim 24, wherein said second type of cell is a hematopoietic stem or progenitor cell.

33. A permanent or degradable decellularized or synthetic matrix or scaffold comprising the cell of any of claim 1, 2, 4, 7 or 9.

34. The matrix or scaffold of claim 33, wherein said matrix or scaffold is an amniotic membrane;

a dehydrated extracellular matrix;

placental collagen, or placental extracellular membrane.

35. A method of treating an individual having a disease or disorder of the circulatory system, comprising administering a population of the cells of claim 1 or claim 7 to said individual in an amount and for a time sufficient for detectable improvement of one or more symptoms of said disease or disorder.

36. A method of treating an individual having a disease or disorder of the circulatory system, comprising administering a population of the cells of claim 1 or claim 7 to said individual in an amount and for a time sufficient for detectable improvement of one or more indicia of cardiac function, wherein said indicia of cardiac function are chest cardiac output (CO), cardiac index (CI), pulmonary artery wedge pressure (PAWP), cardiac index (CI), % fractional shortening (% FS), ejection fraction (EF), left ventricular ejection fraction (LVEF);

left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD), contractility (dP/dt), a decrease in atrial or ventricular functioning, an increase in pumping efficiency, a decrease in the rate of loss of pumping efficiency, a decrease in loss of hemodynamic functioning, or decrease in complications associated with cardiomyopathy, as compared to the individual prior to administration of amnion derived adherent cells.

37. A method of treating an individual having a disruption of blood flow in or around the CNS, comprising administering a therapeutically effective amount of the amnion derived adherent cells of claim 1 or claim 7.

38. A method of treating an individual having a disruption of blood flow in or around a limb, comprising administering a therapeutically effective amount of the amnion derived adherent cells of claim 1 or claim 7.

28. The isolated population of claim 28, wherein said hematopoietic stem or progenitor cell is a CD34⁺ cell.

29. An isolated amnion derived adherent cell, wherein said cell is adherent to tissue culture plastic, and wherein said cell is OCT-4⁻

, as determined by RT-PCR, and CD49f⁺, HLA-G⁻

, CD90⁺, CD105⁺, and CD117⁻

, as determined by immunolocalization, and wherein said cell;

(a) expresses one or more of CD9, CD10, CD44, CD54, CD98, CD200, Tie-2, TEM-7, VEGFR1/Flt-1, or VEGFR2/KDR (CD309), as determined by immunolocalization;

(b) lacks expression of CD31, CD34, CD38, CD45, CD133, CD143, CD144, CD146, CD271, CXCR4, HLA-G, or VE-cadherin, as determined by immunolocalization, or lacks expression of SOX2, as determined by RT-PCR;

(c) express mRNA for ACTA2, ADAMTS1, AMOT, ANG, ANGPT1, ANGPT2, ANGPTL1, ANGPTL2, ANGPTL4, BAI1, CD44, CD200, CEACAM1, CHGA, COL15A1, COL18A1, COL4A1, COL4A2, COL4A3, CSF3, CTGF, CXCL12, CXCL2, DNMT3B, ECGF1, EDG1, EDIL3, ENPP2, EPHB2, FBLN5, F2, FGF1, FGF2, FIGF, FLT4, FN1, FST, FOXC2, GRN, HGF, HEY1, HSPG2, IFNB1, IL8, IL12A, ITGA4, ITGAV, ITGB3, MDK, MMP2, MYOZ2, NRP1, NRP2, PDGFB, PDGFRA, PDGFRB, PECAM1, PF4, PGK1, PROX1, PTN, SEMA3F, SERPINB5, SERPINC1, SERPINF1, TIMP2, TIMP3, TGFA, TGFB1, THBS1, THBS2, TIE1, TIE2/TEK, TNF, TNNI1, TNFSF15, VASH1, VEGF, VEGFB, VEGFC, VEGFR1/FLT1, or VEGFR2/KDR;

(d) expresses one or more of the proteins CD49d, Connexin-43, HLA-ABC, Beta 2-microglobulin, CD349, CD318, PDL1, CD106, Galectin-1, ADAM 17, angiotensinogen precursor, filamin A, alpha-actinin 1, megalin, macrophage acetylated LDL receptor I and II, activin receptor type IIB precursor, Wnt-9 protein, glial fibrillary acidic protein, astrocyte, myosin-binding protein C, or myosin heavy chain, nonmuscle type A;

(e) secretes VEGF, HGF, IL-8, MCP-3, FGF2, Follistatin, G-CSF, EGF, ENA-78, GRO, IL-6, MCP-1, PDGF-BB, TIMP-2, uPAR, or galectin-1 into culture medium in which the cell grows;

(f) expresses micro RNAs miR-17-3p, miR-18a, miR-18b, miR-19b, miR-92, or miR-296 at a higher level than an equivalent number of bone marrow-derived mesenchymal stem cells;

(g) expresses micro RNAs miR-20a, miR-20b, miR-221, miR-222, miR-15b, or miR-16 at a lower level than an equivalent number of bone marrow-derived mesenchymal stem cells;

(h) expresses miRNAs miR-17-3p, miR-18a, miR-18b, miR-19b, miR-92, miR-20a, miR-20b, miR-296, miR-221, miR-222, miR-15b, or miR-16;

or

(i) expresses increased levels of CD202b, IL-8 or VEGF when cultured in less than about 5% O₂, compared to expression of CD202b, IL-8 or VEGF under 21% O₂.

30. The isolated amnion derived adherent cell of claim 29, wherein said cell is OCT-4⁻

, as determined by RT-PCR, and CD49f⁺, HLA-G⁻

, CD90⁺, CD105⁺, and CD117⁻

, as determined by immunolocalization, and wherein said cell;

(a) expresses CD9, CD10, CD44, CD54, CD98, CD200, Tie-2, TEM-7, VEGFR1/Flt-1, and/or VEGFR2/KDR (CD309), as determined by immunolocalization;

(b) lacks expression of CD31, CD34, CD38, CD45, CD133, CD143, CD144, CD146, CD271, CXCR4, HLA-G, and/or VE-cadherin, as determined by immunolocalization, and/or lacks expression of SOX2, as determined by RT-PCR;

(c) express mRNA for ACTA2, ADAMTS1, AMOT, ANG, ANGPT1, ANGPT2, ANGPTL1, ANGPTL2, ANGPTL4, BAI1, CD44, CD200, CEACAM1, CHGA, COL15A1, COL18A1, COL4A1, COL4A2, COL4A3, CSF3, CTGF, CXCL12, CXCL2, DNMT3B, ECGF1, EDG1, EDIL3, ENPP2, EPHB2, FBLN5, F2, FGF1, FGF2, FIGF, FLT4, FN1, FST, FOXC2, GRN, HGF, HEY1, HSPG2, IFNB1, IL8, IL12A, ITGA4, ITGAV, ITGB3, MDK, MMP2, MYOZ2, NRP1, NRP2, PDGFB, PDGFRA, PDGFRB, PECAM1, PF4, PGK1, PROX1, PTN, SEMA3F, SERPINB5, SERPINC1, SERPINF1, TIMP2, TIMP3, TGFA, TGFB1, THBS1, THBS2, TIE1, TIE2/TEK, TNF, TNNI1, TNFSF15, VASH1, VEGF, VEGFB, VEGFC, VEGFR1/FLT1, and/or VEGFR2/KDR;

(d) expresses one or more of CD49d, Connexin-43, HLA-ABC, Beta 2-microglobulin, CD349, CD318, PDL1, CD106, Galectin-1, ADAM 17, angiotensinogen precursor, filamin A, alpha-actinin 1, megalin, macrophage acetylated LDL receptor I and II, activin receptor type IIB precursor, Wnt-9 protein, glial fibrillary acidic protein, astrocyte, myosin-binding protein C, and/or myosin heavy chain, nonmuscle type A;

(e) secretes one or more of VEGF, HGF, IL-8, MCP-3, FGF2, Follistatin, G-CSF, EGF, ENA-78, GRO, IL-6, MCP-1, PDGF-BB, TIMP-2, uPAR, and Galectin-1 into culture medium in which the cell grows;

(f) expresses micro RNAs miR-17-3p, miR-18a, miR-18b, miR-19b, miR-92, and/or miR-296 at a higher level than an equivalent number of bone marrow-derived mesenchymal stem cells;

(g) expresses micro RNAs miR-20a, miR-20b, miR-221, miR-222, miR-15b, and/or miR-16 at a lower level than an equivalent number of bone marrow-derived mesenchymal stem cells; and

(h) expresses miRNAs miR-17-3p, miR-18a, miR-18b, miR-19b, miR-92, miR-20a, miR-20b, miR-296, miR-221, miR-222, miR-15b, and/or miR-16; and

/or

(i) expresses increased levels of CD202b, IL-8 and/or VEGF when cultured in less than about 5% O₂, compared to expression of CD202b, IL-8 and/or VEGF under 21% O₂.

31. An isolated population of the cells of claim 29.

32. An isolated population of the cells of claim 30.