Postpartum cells derived from placental tissue, and methods of making and using the same

• US 10,195,233 B2
• Filed: 02/27/2017
• Issued: 02/05/2019
• Est. Priority Date: 06/27/2003
• Status: Active Grant

• Current Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Original Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Inventors: Kihm, Anthony J., Harris, Ian Ross, Mistry, Sanjay, Harmon, Alexander M., Messina, Darin J., Seyda, Agnieszka, Yi, Chin-Feng, Gosiewska, Anna
• Primary Examiner(s): Fox, Allison M
• Application Number:
  US15/443,718
• Publication Number:
  US 20170166863A1
• Time in Patent Office: 708 Days
• 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: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 38/1808 Epidermal growth factor [EGF] urogastrone
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 38/1833 Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
• CPC Class Codes: A61K 38/1841 Transforming growth factor [TGF]
• CPC Class Codes: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61K 38/1858 Platelet-derived growth factor [PDGF]
• CPC Class Codes: A61K 38/1866 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: A61K 38/1891 Angiogenesic factors; Angiogenin
• CPC Class Codes: A61K 38/19 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 38/204 IL-6
• CPC Class Codes: A61K 38/2053 IL-8
• CPC Class Codes: A61K 38/27 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 13/12 : of the kidneys
• CPC Class Codes: A61P 17/02 : for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 : for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/08 : for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 : for osteoporosis
• CPC Class Codes: A61P 21/00 : Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 : for peripheral neuropathies
• CPC Class Codes: A61P 25/14 : for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 : Anti-Parkinson drugs
• 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 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 27/06 : Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/06 : Free radical scavengers or antioxidants
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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 2500/32 : Amino acids
• CPC Class Codes: C12N 2500/34 : Sugars
• CPC Class Codes: C12N 2500/44 : Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2500/90 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2500/95 : Protein-free medium and culture conditions
• CPC Class Codes: C12N 2501/12 : Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/21 : Chemokines, e.g. MIP-1, MIP-2, RANTES, MCP, PF-4
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• CPC Class Codes: C12N 2502/02 : embryonic cells
• 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 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2533/50 : Proteins
• 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 5/0607 : Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696

Abstract

Cells derived from postpartum placenta and methods for their isolation are provided by the invention. The invention further provides cultures and compositions of the placenta-derived cells. The placenta-derived cells of the invention have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications.

Claims

1. A method of isolating a placenta-derived cell from a human postpartum placenta comprising dissociating said cell from said placenta with a matrix metalloprotease, a neutral protease, and a mucolytic enzyme that digests hyaluronic acid, wherein the isolated placenta-derived cell is obtained from human postpartum placenta substantially free of blood, and wherein the placenta-derived cell has the following characteristics:

a) self-renews and expands in culture;

b) is multipotent;

c) produces CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A,B,C;

d) does not produce CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G or HLA-DR-DP,DQ; and

e) expresses, relative to a human fibroblast, mesenchymal stem cell, or iliac crest bone marrow cell, increased levels of oxidized low density lipoprotein receptor 1 and renin.

2. The method of claim 1, wherein said matrix metalloprotease comprises collagenase.

3. The method of claim 1, wherein said neutral protease comprises dispase.

4. The method of claim 1, wherein said mucolytic enzyme comprises hyaluronidase.

5. The method of claim 1, further comprising mechanically dissociating said placenta prior to said dissociating step.

6. The method of claim 1, further comprising growing said cell in culture medium.

7. The method of claim 6, wherein said culture medium comprises RPMI1640, Ham'"'"'s F10 medium, Ham'"'"'s F12 medium, Mesenchymal Stem Cell Growth Medium, Iscove'"'"'s modified Dulbecco'"'"'s medium, Dulbecco'"'"'s modified Eagle'"'"'s Medium (DMEM), CELL-GRO FREE, DMEM/F12, advanced DMEM, DMEM/MCDB201, or Eagle'"'"'s basal medium.

8. The method of claim 6, wherein said medium comprises about 2% to about 15% (v/v) serum.

9. The method of claim 6, wherein said medium comprises beta-mercaptoethanol.

10. The method of claim 6, wherein said culture medium comprises at least one antibiotic agent.

11. The method of claim 6, wherein said culture medium comprises glucose.

12. The method of claim 6, wherein said culture medium comprises L-valine.

13. The method of claim 6, wherein said culture medium comprises DMEM-low glucose, beta-mercaptoethanol, serum, and an antibiotic agent.

14. The method of claim 1, further comprising cryopreserving said placenta-derived cell.

15. The method of claim 1, further comprising banking said placenta-derived cell.

16. The method of claim 1, further comprising expanding said isolated cells on an uncoated surface.

17. The method of claim 1, further comprising expanding said isolated cells on a coated surface.

18. The method of claim 17, wherein said surface is coated with at least one of gelatin, collagen, fibronectin, laminin, ornithine, vitronectin, or extracellular membrane protein.

19. The method of claim 6, wherein the culture medium is protein-free culture medium.

20. The method of claim 6, wherein the culture medium is serum-free culture medium.

21. The method of claim 1, wherein the placenta-derived cell lacks production of GRO-alpha and oxidized low density lipoprotein receptor, as detected by flow cytometry.

22. The method of claim 1, wherein the placenta-derived cell further has the following characteristics:

(a) secretion of monocyte chemotactic protein 1 (MCP-1), interleukin-6 (IL-6), interleukin 8 (IL8), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), heparin-binding epidermal growth factor (HB-EGF), brain-derived neurotrophic factor (BDNF), tissue inhibitor of matrix metalloproteinase 1 (TIMP1), thrombopoietin (TPO), macrophage inflammatory protein 1alpha(MIP1a), Rantes (regulated on activation, normal T cell expressed and secreted), thymus and activation-regulated chemokine (TARC), and Eotaxin; and

(b) lack of secretion of fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), angiopoietin 2 (ANG2), platelet derived growth factor (PDGF-bb), transforming growth factor beta2 (TGFbeta2), macrophage inflammatory protein 1beta (MIP1b), I309, and macrophage-derived chemokine (MDC), as detected by ELISA.

23. The method of claim 1, wherein the placenta-derived cell is of a neonatal origin.

24. The method of claim 1, wherein said placenta-derived cell is of a maternal origin.

25. The method of claim 1, wherein the placenta-derived cell has the ability to differentiate into a mesodermal, ectodermal, or endodermal phenotype.

26. The method of claim 1, wherein the placenta-derived cell can undergo at least 40 population doublings in culture.

27. The method claim 1, further comprising genetically engineering the isolated cell to produce a protein of interest.

28. The method of claim 1, wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6074.

29. The method of claim 1, wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6075.

30. The method of claim 1, wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6079.

31. The method of claim 1, wherein the isolated placenta-derived cell expresses granulocyte chemotactic protein-2 (GCP-2).