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

Abstract

Cells derived from human umbilical cords are disclosed along with methods for their therapeutic use. Isolation techniques, culture methods and detailed characterization of the cells with respect to their cell surface markers, gene expression, and their secretion of trophic factors are described.

1 . An isolated umbilicus-derived cell comprising a cell derived from mammalian umbilical cord tissue substantially free of blood, said cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes.  
     
     
         2 . The isolated cell of  claim 1  which can expand in the presence of oxygen from about 5% to about 20%.  
     
     
         3 . The cell of  claim 2  which requires L-valine for growth.  
     
     
         4 . The isolated cell of  claim 3  which can double sufficiently to generate yields of greater than about 10 14  cells in less than about 80 days in culture when seeded at about 10 3  cells/cm 2 .  
     
     
         5 . The isolated cell of  claim 3  which can double sufficiently to generate greater than about 10 15  cells in less than about 80 days in culture when seeded at about 5×10 3  cells/cm 2 .  
     
     
         6 . The isolated cell of  claim 5  which can double sufficiently to generate greater than about 10 17  cells in less than about 65 days in culture when seeded at about 5×10 3  cells/cm 2 .  
     
     
         7 . The isolated cell of  claim 3  which can undergo at least 40 doublings in culture.  
     
     
         8 . The cell of  claim 3  which is isolated from human umbilicus.  
     
     
         9 . The isolated cell of  claim 8  isolated in the presence of one or more enzyme activities comprising metalloproteases, neutral proteases, or mucolytic enzymes.  
     
     
         10 . The isolated cell of  claim 9  wherein the enzyme activities include at least one collagenase, and one or more of the protease activities, dispase and thermolysin.  
     
     
         11 . The isolated cell of  claim 9  wherein the enzyme activities are a collagenase from  Clostridium histolyticum  and dispase.  
     
     
         12 . The isolated cell of  claim 11  wherein the enzyme activities further include hyaluronidase.  
     
     
         13 . The isolated cell of  claim 12  which attaches and expands on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating with gelatin, laminin, collagen, polyornithine, polylysine, vitronectin, or fibronectin.  
     
     
         14 . The isolated cell of  claim 13  which expands in the presence of from about 2% to about 15% added serum, in the presence or absence of beta-mercaptoethanol, and in the presence or absence of added growth factors including EGF, FGF, PDGF, VEGF, IGF or LIF.  
     
     
         15 . A cell culture comprising the isolated cell of  claim 1  which is free of maternal cells.  
     
     
         16 . The isolated cell of  claim 1  which maintains a normal karyotype as it is passaged.  
     
     
         17 . The isolated cell of  claim 1  wherein the cell is characterized in its production or lack of production of one or more cell surface markers comprising CD10, CD13, CD31, CD44, CD45, CD73, CD90, CD117, CD141, PDGFr-alpha, HLA-A, B, C, and HL-Dr, DP, DQ.  
     
     
         18 . The isolated cell of  claim 17  wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A, B, C.  
     
     
         19 . The isolated cell of  claim 18  wherein the cell produces each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A, B, C.  
     
     
         20 . The isolated cell of  claim 17  wherein the cell does not produce one or more of CD31, CD34, CD45, CD117, CD141, or HLA-DR, DP, DQ, as detected by flow cytometry.  
     
     
         21 . The isolated cell of  claim 20  wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A, B, C.  
     
     
         22 . The isolated cell of  claim 20  wherein the cell does not produce any of CD31, CD34, CD45, CD117, CD141, or HLA-DR, DP, DQ, as detected by flow cytometry.  
     
     
         23 . The isolated cell of  claim 22  wherein the cell produces each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A, B, C.  
     
     
         24 . An isolated CD45 −  umbilicus-derived cell having a cell surface marker profile wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A, B, C, and does not express one or more of CD31, CD34, CD117, CD141, or HLA-DR, DP, DQ, as detected by flow cytometry.  
     
     
         25 . The isolated cell of  claim 24  wherein the cell surface marker expression profile remains substantially unchanged with passage, culture vessel surface coating, or isolation procedure.  
     
     
         26 . The isolated cell of  claim 25  expressing a gene for one or more of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; or tumor necrosis factor, alpha-induced protein 3.  
     
     
         27 . The isolated cell of  claim 26  expressing a gene for each of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; and tumor necrosis factor, alpha-induced protein 3.  
     
     
         28 . The isolated cell of  claim 27  wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, an ileac crest bone marrow cell, or placenta-derived cell.  
     
     
         29 . The isolated cell of  claim 25 , which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of one or more genes selected from the group consisting of: short stature homeobox 2; heat shock 27 kDa protein 2; chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);  Homo sapiens  mRNA; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2 (growth arrest-specific homeobox); sine oculis homeobox homolog 1 ( Drosophila ); crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; B-cell translocation gene 1, anti-proliferative; cholesterol 25-hydroxylase; runt-related transcription factor 3; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; frizzled homolog 7 ( Drosophila ); hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C (hexabrachion); iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; early growth response 3; distal-less homeobox 5; hypothetical protein FLJ20373; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglycan; fibronectin 1; proenkephalin; integrin, beta-like 1 (with EGF-like repeat domains);  Homo sapiens  mRNA full length insert cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIa polypeptide 1 (muscle); neuroblastoma, suppression of tumorigenicity 1; and insulin-like growth factor binding protein 2, 36 kDa.  
     
     
         30 . The isolated cell of  claim 29  expressing a gene for one or more of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; or tumor necrosis factor, alpha-induced protein 3, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, an ileac crest bone marrow cell, or placenta-derived cell.  
     
     
         31 . The isolated cell of  claim 29 , which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of genes for each of: short stature homeobox 2; heat shock 27 kDa protein 2; chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);  Homo sapiens  mRNA; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2 (growth arrest-specific homeobox); sine oculis homeobox homolog 1 ( Drosophila ); crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; B-cell translocation gene 1, anti-proliferative; cholesterol 25-hydroxylase; runt-related transcription factor 3; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; frizzled homolog 7 ( Drosophila ); hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C (hexabrachion); iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; early growth response 3; distal-less homeobox 5; hypothetical protein FLJ20373; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglycan; fibronectin 1; proenkephalin; integrin, beta-like 1 (with EGF-like repeat domains);  Homo sapiens  mRNA full length insert cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIa polypeptide 1 (muscle); neuroblastoma, suppression of tumorigenicity 1; and insulin-like growth factor binding protein 2, 36 kDa.  
     
     
         32 . The isolated cell of  claim 31  expressing a gene for each of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; and tumor necrosis factor, alpha-induced protein 3, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell.  
     
     
         33 . An isolated human umbilicus-derived cell, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of genes for each of: short stature homeobox 2; heat shock 27 kDa protein 2; chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);  Homo sapiens  mRNA; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2 (growth arrest-specific homeobox); sine oculis homeobox homolog 1 ( Drosophila ); crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; B-cell translocation gene 1, anti-proliferative; cholesterol 25-hydroxylase; runt-related transcription factor 3; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; frizzled homolog 7 ( Drosophila ); hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C (hexabrachion); iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; early growth response 3; distal-less homeobox 5; hypothetical protein FLJ20373; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglycan; fibronectin 1; proenkephalin; integrin, beta-like 1 (with EGF-like repeat domains);  Homo sapiens  mRNA full length insert cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIa polypeptide 1 (muscle); neuroblastoma, suppression of tumorigenicity 1; insulin-like growth factor binding protein 2, 36 kDa; and which expresses a gene for each of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; and tumor necrosis factor, alpha-induced protein 3, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell.  
     
     
         34 . The isolated cell of  claim 33  capable of self-renewal and expansion in culture, and having the potential to differentiate into cells of other phenotypes.  
     
     
         35 . The isolated cell of  claim 34  which produces one or both of vimentin and alpha smooth muscle actin.  
     
     
         36 . The isolated cell of  claim 35  which produces both vimentin and alpha-smooth muscle actin.  
     
     
         37 . The isolated cell of  claim 36  wherein the production of vimentin and alpha smooth muscle actin is retained over passaging under growth conditions.  
     
     
         38 . A therapeutic cell culture comprising the cell of  claim 34 .  
     
     
         39 . The therapeutic cell culture of  claim 38  which does not substantially stimulate allogeneic PBMCs.  
     
     
         40 . The therapeutic cell culture of  claim 39  which lacks detectable amounts of HLA-DR, HLA-DP, HLA-DQ, CD80, CD86, and B7-H2, as determined by flow cytometry.  
     
     
         41 . The therapeutic cell culture of  claim 40  which further lacks detectable amounts of HLA-G and CD178, as determined by flow cytometry.  
     
     
         42 . The therapeutic cell culture of  claim 41 , which produces detectable amounts of PD-L2, as determined by flow cytometry.  
     
     
         43 . The therapeutic cell culture of  claim 34  which does not substantially stimulate a lymphocyte mediated response in vitro, as compared to allogeneic controls in a mixed lymphocyte reaction.  
     
     
         44 . An isolated human umbilicus-derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes, wherein the cell does not substantially stimulate naïve CD4 +  T cells, and expresses PD-L2, but not HLA-G, CD178, HLA-DR, HLA-DP, HLA-DQ, CD80, CD86, or B7-H2.  
     
     
         45 . The isolated cell of  claim 44  wherein the cell produces vimentin and alpha-smooth muscle actin.  
     
     
         46 . The isolated cell of  claim 44  wherein the cell secretes one or more cellular factors.  
     
     
         47 . The isolated cell of  claim 46  wherein the factors are MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.  
     
     
         48 . The isolated cell of  claim 47  wherein the cell secretes each of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.  
     
     
         49 . The isolated cell of  claim 44  wherein the cell does not secrete one or more of the cellular factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.  
     
     
         50 . The isolated cell of  claim 49 , wherein the cell secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.  
     
     
         51 . The isolated cell of  claim 50  wherein the cell secretes each of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.  
     
     
         52 . The isolated cell of  claim 49  wherein the cell does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.  
     
     
         53 . The isolated cell of  claim 52 , wherein the cell secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.  
     
     
         54 . The isolated cell of  claim 53  wherein the cell secretes each of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.  
     
     
         55 . An isolated human umbilicus-derived cell which secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1, and does not secrete one or more of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.  
     
     
         56 . The isolated cell of  claim 55  which secretes each of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.  
     
     
         57 . The isolated cell of  claim 50  wherein the cell does not secrete any of the factors SD F-1 alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.  
     
     
         58 . A therapeutic cell culture comprising the isolated cell of  claim 55 .  
     
     
         59 . The therapeutic cell culture of  claim 58  further comprising one or more of a pharmaceutically-acceptable carrier, another cell culture, an antiapoptotic compound, an antithrombogenic compound, an anti-inflammatory compound, an immunosuppressive compound, an immunomodulatory compound, an angiogenic factor, and a neurotrophic factor.  
     
     
         60 . A method of deriving, from umbilical tissue, an isolated cell, said cell capable of self-renewal and expansion in culture, and having the potential to differentiate into cells of other phenotypes, the method comprising the steps of: 
 (a) obtaining umbilical tissue;    (b) removing substantially all of blood to yield a substantially blood-free umbilical tissue,    (c) dissociating the tissue by mechanical or enzymatic treatment, or both,    (d) resuspending the tissue in a culture medium, and    (e) providing growth conditions which allow for the growth of an umbilicus-derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes.    
     
     
         61 . The method of  claim 60  further comprising the step of selecting adherent cells after from about ten to about 100 hours in culture.  
     
     
         62 . The method of  claim 61  wherein the umbilicus tissue is from a human.  
     
     
         63 . The method of  claim 62  wherein the derived cell is capable of to expansion in the absence of added growth factors in medium with from 2% to about 15% added serum.  
     
     
         64 . The method of  claim 63  wherein the derived cell can expand in the presence of oxygen from about 5% to about 20%.  
     
     
         65 . The method of  claim 64  wherein the derived cell cannot be maintained in the absence of L-valine.  
     
     
         66 . The method of  claim 65  wherein the derived cell can undergo at least 40 doublings in culture.  
     
     
         67 . The method of  claim 65  wherein the derived cell can double sufficiently to generate at least about 10 17  cells in less than about 65 days in culture when seeded at about 5×10 3  cells/cm 2 .  
     
     
         68 . The method of  claim 62  wherein the umbilical tissue is obtained after normal or surgically-assisted child birth from a full-term or pre-term pregnancy.  
     
     
         69 . The method of  claim 60  wherein the dissociation step comprises the use of one or more enzyme activities selected from the group consisting of metalloprotease, hyaluronidase, and neutral protease.  
     
     
         70 . The method of  claim 69  wherein the enzyme activities are collagenase and dispase.  
     
     
         71 . The method of  claim 70  wherein the enzyme activities further include hyaluronidase.  
     
     
         72 . The method of  claim 71  wherein the dissociating step comprises incubating at about 37° C.  
     
     
         73 . The method of  claim 72  wherein the incubating is for one or more hours.  
     
     
         74 . The method of  claim 72  where the incubating is for about 2 hours.  
     
     
         75 . The method of  claim 69  wherein the derived cell attaches and expands on a coated or uncoated tissue culture vessel, wherein the coated tissue culture vessel comprises a coating with gelatin, laminin, collagen, polyornithine, polylysine, vitronectin, or fibronectin.  
     
     
         76 . The method of  claim 69  wherein the derived cell expands in the presence of from about 2% to about 15% Fetal Bovine Serum, in the presence or absence of beta-mercaptoethanol, and in the presence or absence of one or more added growth factors including EGF, FGF, PDGF, VEGF, IGF and LIF.  
     
     
         77 . The method of  claim 60  wherein the removing step comprises removal of free or clotted blood by one or more of washing, suctioning, blotting, centrifugal separation, or enzymatic removal.  
     
     
         78 . The method of  claim 60  wherein the dissociating step is accomplished aseptically.  
     
     
         79 . The method of  claim 78  wherein the dissociation step comprises one or more of mincing, blending, homogenizing, or grinding.  
     
     
         80 . An isolated human umbilicus-derived cell derived by the method of  claim 69 .  
     
     
         81 . The isolated cell of  claim 80  which maintains a normal karyotype with passaging.  
     
     
         82 . A therapeutic culture of human umbilicus-derived cells derived by the method of  claim 69 , wherein the culture is free of maternal cells.  
     
     
         83 . A conditioned culture medium generated by the growth of the culture of  claim 15  or  82 .  
     
     
         84 . The conditioned medium of  claim 83  comprising one or more of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.  
     
     
         85 . A mammalian cell culture comprising the conditioned medium of  claim 84  and a mammalian cell in need of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.  
     
     
         86 . The culture of  claim 15  or  claim 82  comprising the umbilicus-derived cells and another mammalian cell of any phenotype.  
     
     
         87 . The culture of claim of  claim 86  comprising a human cell line in addition to the umbilicus-derived cell.  
     
     
         88 . A three dimensional matrix comprising the umbilicus-derived cell of  claim 1 .  
     
     
         89 . The three-dimensional matrix of  claim 88  wherein the matrix comprises a biocompatible or bioabsorbable polymer.  
     
     
         90 . The three-dimensional matrix of  claim 89  wherein the matrix comprises PGA/PLA copolymer, PCL/PGA copolymer, or self assembling peptides.  
     
     
         91 . An implantable tissue structure comprising the matrix of  claim 89 .  
     
     
         92 . An implantable device comprising the therapeutic cell of  claim 82 .  
     
     
         93 . An implantable human tissue matrix comprising the cell of  claim 80 .  
     
     
         95 . A human tissue comprising the cell of  claim 8 .  
     
     
         96 . A cell lysate derived from the cell of  claim 8 .  
     
     
         97 . A soluble cell fraction derived from the cell of  claim 8   
     
     
         98 . A membrane-enriched cell fraction derived from the cell of  claim 8 .  
     
     
         99 . An extracellular membrane fraction derived from the cell of  claim 8 .  
     
     
         100 . An injectable therapeutic cell comprising an isolated human umbilicus-derived cell of  claim 44 .  
     
     
         101 . The injectable cell of  claim 100  treated to inactivate tissue factor.  
     
     
         102 . The injectable cell of  claim 101  wherein the treated is with an anti-tissue factor antibody.  
     
     
         103 . An isolated postpartum-derived cell comprising an L-valine-requiring cell derived from human postpartum tissue substantially free of blood, said cell capable of self-renewal and expansion in culture and having the potential to differentiate into a cell of cardiomyocyte phenotypes; the cell capable of growth in an atmosphere containing oxygen from about 5% to at least about 20%; wherein said cell comprises at least one of the following characteristics: 
 potential for at least about 40 doublings in culture;    attachment and expansion on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyornithine, vitronectin, or fibronectin;    production of at least one of tissue factor, vimentin, and alpha-smooth muscle actin;    production of at least one of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A, B, C;    lack of production of at least one of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, and HLA-DR, DP, DQ, as detected by flow cytometry;    expression of at least one of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; and tumor necrosis factor, alpha-induced protein 3;    expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is reduced for at least one of: short stature homeobox 2; heat shock 27 kDa protein 2; chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);  Homo sapiens  mRNA; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2 (growth arrest-specific homeobox); sine oculis homeobox homolog 1 ( Drosophila ); crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; B-cell translocation gene 1, anti-proliferative; cholesterol 25-hydroxylase; runt-related transcription factor 3; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; frizzled homolog 7 ( Drosophila ); hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C (hexabrachion); iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; early growth response 3; distal-less homeobox 5; hypothetical protein FLJ20373; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglycan; fibronectin 1; proenkephalin; integrin, beta-like 1 (with EGF-like repeat domains);  Homo sapiens  mRNA full length insert cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIa polypeptide 1 (muscle); neuroblastoma, suppression of tumorigenicity 1; insulin-like growth factor binding protein 2, 36 kDa;    secretion of at least one of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1a, RANTES, and TIMP1; and    lack of secretion of at least one of TGF-beta2, ANG2, PDGFbb, MIP1beta, 1309, MDC, and VEGF, as detected by ELISA.    
     
     
         104 . The isolated postpartum-derived cell of  claim 103;  wherein said cell comprises each of the following characteristics: 
 potential for at least about 40 doublings in culture;    attachment and expansion on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyornithine, vitronectin, or fibronectin;    production of tissue factor, vimentin, and alpha-smooth muscle actin;    production of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A, B, C;    lack of production of each of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, and HLA-DR, DP, DQ, as detected by flow cytometry;    expression of interleukin 8; reticulon 1; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha); chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); chemokine (C-X-C motif) ligand 3; and tumor necrosis factor, alpha-induced protein 3;    expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is reduced for each of: short stature homeobox 2; heat shock 27 kDa protein 2; chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);  Homo sapiens  mRNA; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2 (growth arrest-specific homeobox); sine oculis homeobox homolog 1 ( Drosophila ); crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; B-cell translocation gene 1, anti-proliferative; cholesterol 25-hydroxylase; runt-related transcription factor 3; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; frizzled homolog 7 ( Drosophila ); hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C (hexabrachion); iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; early growth response 3; distal-less homeobox 5; hypothetical protein FLJ20373; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglycan; fibronectin 1; proenkephalin; integrin, beta-like 1 (with EGF-like repeat domains);  Homo sapiens  mRNA full length insert cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIa polypeptide 1 (muscle); neuroblastoma, suppression of tumorigenicity 1; insulin-like growth factor binding protein 2, 36 kDa;    secretion of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1a, RANTES, and TIMP1; and    lack of secretion of each of TGF-beta2, ANG2, PDGFbb, MIP1beta, 1309, MDC, and VEGF, as detected by ELISA.    
     
     
         105 . An isolated postpartum-derived cell comprising a signature gene profile wherein mRNA from genes for reticulon, oxidized LDL receptor, and IL-8 are present independent of whether the cells are grown in medium containing serum or medium free of serum.  
     
     
         106 . The isolated postpartum-derived cell of  claim 105  further comprising the ability to alter its expression of cell surface markers when grown in medium containing serum relative to that in serum free medium.  
     
     
         107 . The isolated postpartum-derived cell of  claim 106  wherein the markers for PDGFreceptor alpha and HLA-ABC are altered.  
     
     
         108 . A method for the preparation of therapeutic cells or cultures comprising: 
 a) isolating cells;    b) initially expanding the cells to a useful number in a serum-containing medium which supports cell expansion but in which the cells produce a quantity of the cell surface marker HLA-ABC;    c) transferring the cells to a medium in which the cells produce a decreased amount of the cell surface marker HLA-ABC; and    d) passaging the cells in the medium in which the cells produce a decreased amount of the HLA-ABC,    thereby preparing a therapeutic cell or culture.    
     
     
         109 . The method of  claim 108  wherein the medium in which the cells produce a decreased amount of the cell surface marker HLA-ABC is a serum-free medium.  
     
     
         110 . The method of  claim 109  for the production of therapeutic cells or cultures for implantation.  
     
     
         111 . The method of  claim 108  for the production of therapeutic cells or cultures for grafting.  
     
     
         112 . A serum-free medium for the expansion of postpartum-derived cells wherein the medium has one or more growth factors added.  
     
     
         113 . The serum-free medium of  claim 112  wherein the one or more growth factors added are bFGF, EGF, or PDGF.  
     
     
         114 . The serum-free medium of  claim 113  wherein the one or more added growth factors includes bFGF.  
     
     
         115 . The serum free medium of  claim 114  which supports expansion for at least 20 passages.  
     
     
         116 . A therapeutic culture comprising postpartum-derived cells expanded in serum-free medium.  
     
     
         117 . The therapeutic culture of  claim 116  comprising cells having a signature gene profile wherein mRNA from genes for reticulon, oxidized LDL receptor, and IL-8 are present independent of whether the cells are grown in medium containing serum or medium free of serum.  
     
     
         118 . A cell culture bank comprising the cells of 117.