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

• US 9,579,351 B2
• Filed: 06/25/2004
• Issued: 02/28/2017
• 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:
  US10/877,446
• Publication Number:
  US 20050058631A1
• Time in Patent Office: 4,631 Days
• US Class Current: 1/1
• 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 cell culture comprising an isolated homogeneous cell population and a culture medium, wherein said isolated cell population is obtained from human postpartum placenta substantially free of blood, wherein said cell population self-renews and expands in culture;

is multipotent;

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

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

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

produces granulocyte chemotactic protein-2 (GCP-2); and

has the ability to undergo at least 40 population doublings in culture, and wherein the culture medium comprises Dulbecco'"'"'s modified Eagle'"'"'s medium (DMEM)-low glucose, DMEM-high glucose, RPMI1640, serum- and protein-free DMEM, DMEM/MCDB201, Ham'"'"'s F10 medium, Ham'"'"'s F12 medium, DMEM/F12, Iscove'"'"'s modified Dulbecco'"'"'s medium, or Eagle'"'"'s basal medium.

2. The cell of claim 1 wherein said cell lacks production of GRO-alpha and oxidized low density lipoprotein receptor, as detected by flow cytometry.

3. The cell culture of claim 1 wherein said cell population 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 1 alpha(MIP 1 a), 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.

4. The cell of claim 1 wherein said cell population is of a neonatal origin.

5. The cell of claim 1 wherein said cell population is of a maternal origin.

6. The cell culture of claim 1 wherein the culture medium comprises serum.

7. The cell of claim 6 wherein said culture medium comprises 2% -15% (v/v) serum.

8. The cell culture of claim 7 further comprising at least one growth factor of platelet derived growth factor-bb, epidermal growth factor, vascular endothelial growth factor, fibroblast growth factor, or leukemia inhibitory factor.

9. The cell culture of claim 6 wherein said culture medium comprises at least one of fibroblast growth factor, platelet-derived growth factor, vascular endothelial growth factor, and epidermal growth factor.

10. The culture of claim 6 that can undergo at least 40 population doublings in culture.

11. The cell culture of claim 1 wherein said culture medium comprises betamercaptoethanol.

12. The cell culture of claim 1 wherein said culture medium comprises at least one antibiotic agent.

13. The cell culture of claim 1 wherein said cell population expands in the following culture media in order of growth:

Growth medium>

Mesenchymal Stem Cell Growth Medium (MSCGM)>

Iscove'"'"'s+10% serum=DMEM-high glucose+10% serum=Ham'"'"'s F12+10% serum=RPMI1640+10% serum.

14. The cell culture of claim 1 wherein said culture medium is protein-free culture medium.

15. The cell culture of claim 1 wherein said culture medium is serum-free culture medium.

16. The cell culture of claim 1 wherein said cell population cannot expand in the absence of L-valine.

17. The cell culture of claim 7 wherein the cell population has the ability to differentiate into a mesodermal, ectodermal, or endodermal phenotype.

18. The cell culture of claim 1 further comprising at least one growth factor of platelet derived growth factor-bb, epidermal growth factor, vascular endothelial growth factor, fibroblast growth factor, or leukemia inhibitor factor.

19. The cell culture of claim 1 wherein the culture medium further comprises at least one antimycotic agent.

20. The cell culture of claim 1 wherein said cell population characteristics are stable with passage of said cell population.

21. The cell culture of claim 1 wherein the cell population is genetically engineered to produce a protein of interest.

22. The cell culture of claim 1, wherein the cell population has undergone at least one doubling in culture.

23. The cell culture of claim 1 wherein said cell population obtained from human postpartum placenta substantially free of blood is produced by contacting said human postpartum placenta substantially free of blood with a composition comprising a matrix metalloprotease and a neutral protease and obtaining the cells yielded by said contacting.

24. The cell culture of claim 23 wherein said composition further comprises a mucolytic enzyme that digests hyaluronic acid.

25. The cell culture of claim 1 wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6074.

26. The cell culture of claim 1 wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6075.

27. The cell culture of claim 1 wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6079.

28. A cell culture comprising an isolated homogeneous cell population and a culture medium, wherein said isolated cell population is obtained from human postpartum placenta substantially free of blood, wherein said cell population self-renews and expands in culture;

is multipotent;

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

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

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

produces granulocyte chemotactic protein-2 (GCP-2); and

has the ability to undergo at least 40 population doublings in culture.

29. A matrix comprising a cell culture,

wherein the matrix comprises a nonwoven scaffold comprising a copolymer of glycolic and lactic acids, a 35/65 PCL/PGA foam, an in situ polymerizable gel, or a self-assembling peptide hydrogel, and

wherein the cell culture comprises an isolated homogeneous cell population and a culture medium, wherein said isolated cell population is obtained from human postpartum placenta substantially free of blood, wherein said cell population self-renews and expands in culture;

is multipotent;

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

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

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

produces granulocyte chemotactic protein-2 (GCP-2).

30. The matrix of claim 29 wherein said matrix comprises a nonwoven scaffold comprising a copolymer of glycolic and lactic acids.

31. The matrix of claim 29 wherein the matrix comprises a self-assembling peptide hydrogel.

32. A kit comprising the cell culture comprising an isolated homogeneous cell population, a culture medium and at least one additional component selected from the group consisting of a scaffold, a hydrating agent, and a cell culture substrate, wherein the cell population is cryopreserved, and wherein said isolated cell population is obtained from human postpartum placenta substantially free of blood, wherein said cell population self-renews and expands in culture;

is multipotent;

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

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

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

produces granulocyte chemotactic protein-2 (GCP-2).

33. The kit of claim 32 wherein said cell culture is seeded on said scaffold.

34. A cryopreserved composition comprising an isolated homogeneous cell population, wherein said isolated cell population is obtained from human postpartum placenta substantially free of blood, wherein said cell population self-renews and expands in culture;

is multipotent;

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

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

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

produces granulocyte chemotactic protein-2 (GCP-2); and

has the ability to undergo at least 40 population doublings in culture.

35. The cryopreserved composition of claim 34, wherein said cell population lacks production of GRO-alpha and oxidized low density lipoprotein receptor, as detected by flow cytometry.

36. The cryopreserved composition of claim 34, wherein said cell population 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 1 alpha(MIP 1a), 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.

37. The cryopreserved composition of claim 34, further comprising a matrix, wherein the matrix comprises a nonwoven scaffold comprising a copolymer of glycolic and lactic acids, a 35/65 PCL/PGA foam, an in situ polymerizable gel, or a self-assembling peptide hydrogel.

38. The cryopreserved composition of claim 37, wherein said matrix comprises a nonwoven scaffold comprising a copolymer of glycolic and lactic acids.

39. The cryopreserved composition of claim 37, wherein the matrix comprises a self- assembling peptide hydrogel.

40. The cryopreserved composition of claim 34, wherein said cell population obtained from human postpartum placenta substantially free of blood is produced by contacting said human postpartum placenta substantially free of blood with matrix metalloprotease enzyme and a neutral protease enzyme and obtaining the cells yielded by said contacting.

41. The cryopreserved composition of claim 40, further comprising contacting said human postpartum placenta with a mucolytic enzyme that digests hyaluronic acid.

42. The cryopreserved composition of claim 34, wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6074.

43. The cryopreserved composition of claim 34, wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6075.

44. The cryopreserved composition of claim 34, wherein the isolated homogeneous cell population is identified by ATCC Accession No. PTA-6079.