Postpartum cells derived from placental tissue, and methods of making and using the same
View Patent ↗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.
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, B 7 -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.