Postpartum-derived cells for use in treatment of disease of the heart and circulatory system

Abstract

Cells derived from postpartum tissue are disclosed along with methods for their therapeutic use in diseases of the heart or circulatory system are disclosed. Cells may be used therapeuticall in either differentiated or undifferentiated forms, in homogenous cultures, or as populations with other cells, and in conjunction with other bioactive factors.

1 . 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 of at least one of C-type (calcium dependent, carbohydrate-recognition domain) lectin, superfamily member 2 (activation-induced); Wilms tumor 1; aldehyde dehydrogenase 1 family, member A2; and renin; oxidized low density lipoprotein (lectin-like) receptor 1;  Homo sapiens,  clone IMAGE:4179671, mRNA, partial cds; protein kinase C, zeta; hypothetical protein DKFZp564FO13; downregulated in ovarian cancer 1;  Homo sapiens  mRNA; and cDNA DKFZp547K1113 (from clone DKFZp547K1113);    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 FLJ 14054;  Homo sapiens  mRNA; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; BCL2/adenovirus EIB 19 kDa interacting protein 3-like; AE binding protein 1; cytochrome c oxidase subunit VIIIa 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, MIP1b, 1309, MDC, and VEGF, as detected by ELISA.    
     
     
         2 . The postpartum-derived cell of  claim 1  isolated in the presence of one or more enzyme activities comprising metalloprotease activity, mucolytic activity and neutral protease activity.  
     
     
         3 . The postpartum-derived cell of  claim 2  wherein the enzyme activities are collagenase and dispase.  
     
     
         4 . The postpartum-derived cell of  claim 3  further comprising hyaluronidase.  
     
     
         5 . The postpartum-derived cell of  claim 4  comprising a normal karyotype.  
     
     
         6 . The postpartum-derived cell of  claim 5  that maintains its karyotype as it is passaged.  
     
     
         7 . The postpartum-derived cell of  claim 6  wherein the cell expresses each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A,B,C.  
     
     
         8 . The postpartum-derived cell of  claim 7  wherein the cell does not express any of CD31, CD34, CD45, CD1 17, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.  
     
     
         9 . The postpartum-derived cell of  claim 8  wherein the cell does not spontaneously differentiate along a cardiogenic, angiogenic, hemangiogenic, or vasculogenic pathway when cultured in Growth Medium.  
     
     
         10 . A population of cells comprising the postpartum-derived cell of  claim 8 .  
     
     
         11 . The population of  claim 10  comprising from about 1% postpartum-derived cells to about 10% postpartum cells.  
     
     
         12 . The population of  claim 11  comprising about 50% postpartum-derived cells.  
     
     
         13 . The population of  claim 12  comprising at least 90% postpartum-derived cells.  
     
     
         14 . The population of  claim 13  comprising substantially only postpartum-derived cells.  
     
     
         15 . The population of  claim 14  comprising a clonal cell line of postpartum-derived cells.  
     
     
         16 . A cell lysate prepared from the population of  claim 10 .  
     
     
         17 . The population of  claim 10  incubated in the presence of one or more factors which stimulate stem cell differentiation along a cardiogenic, angiogenic, hemangiogenic, or vasculogenic pathway.  
     
     
         18 . The population of  claim 17  wherein the factors comprise at least one of a demethylation agent, a member of BMP, FGF, TAK, GATA, Csx, NK, MEF2, ET-1, and Wnt factor families, Hedgehog, Csx/Nkx-2.5, and anti-Wnt factors.  
     
     
         19 . The population of  claim 18  wherein the demethylation agent comprises an inhibitor of DNA methyltransferase or an inhibitors of a histone deacetylase, or inhibitors of a repressor complex.  
     
     
         20 . The population of  claim 18  wherein the demethylation agents comprise at least one of 5-azacytidine, 5-aza-2′-deoxycytidine, DMSO, chelerythrine chloride, retinoic acid or salts thereof, 2-amino-4-(ethylthio)butyric acid, procainamide, and procaine.  
     
     
         21 . The population of  claim 10  that is seed onto a matrix to form a matrix-cell complex.  
     
     
         22 . The population of  claim 21  wherein the matrix is a scaffold.  
     
     
         23 . The population of  claim 22  wherein the scaffold is bioabsorbable.  
     
     
         24 . The population of  claim 23  wherein the scaffold comprises at least one other cell type.  
     
     
         25 . The population of  claim 24  wherein the other cell type is a stem cell.  
     
     
         26 . A therapeutic cell composition comprising a pharmaceutically-acceptable carrier and post-partum-derived cells derived from human postpartum tissue substantially free of blood, said cells capable of self-renewal and expansion in culture and having the potential to differentiate into cells of cardiomyocyte phenotypes; the cells capable of growth in an atmosphere containing oxygen from about 5% to at least about 20%; wherein said cells: 
 require L-valine for growth;    have the potential for at least about 40 doublings in culture;    attach and expand on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyomithine, vitronectin, or fibronectin;    produce tissue factor, vimentin, and alpha-smooth muscle actin;    produce each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A,B,C; and    do not produce any of CD31, CD34, CD45, CD117, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.    
     
     
         27 - 40 . (canceled)  
     
     
         41 . The therapeutic cell composition of  claim 26  further comprising the following characteristics: 
 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 of at least one of C-type (calcium dependent, carbohydrate-recognition domain) lectin, superfamily member 2 (activation-induced); Wilms tumor 1; aldehyde dehydrogenase 1 family, member A2; and renin; oxidized low density lipoprotein (lectin-like) receptor 1;  Homo sapiens,  clone IMAGE:4179671, mRNA, partial cds; protein kinase C, zeta; hypothetical protein DKFZp564F013; downregulated in ovarian cancer 1;  Homo sapiens  mRNA; and cDNA DKFZp547K1113 (from clone DKFZp547K1113);    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; disheveled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin (plasminogen binding protein); src homology three (SH3) and cysteine rich domain; cholesterol 25-hydroxylase; runt-related transcription factor 3; 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; neuroblastoma, suppression of tumorigenicity 1; insulin-like growth factor binding protein 2, 36 kDa;  Homo sapiens  cDNA FLJ12280 fis, clone MAMMA1001744; cytokine receptor-like factor 1; potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4; integrin, beta 7; transcriptional co-activator with PDZ-binding motif (TAZ); sine oculis homeobox homolog 2 ( Drosophila ); KIAA1034 protein; vesicle-associated membrane protein 5 (myobrevin); EGF-containing fibulin-like extracellular matrix protein 1; 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; transcriptional co-activator with PDZ-binding motif (TAZ); 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); BCL2/adenovirus E1B 19 kDa interacting protein 3-like; AE binding protein 1; and cytochrome c oxidase subunit VIla polypeptide 1 (muscle);    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, MIP1b, 1309, MDC, and VEGF, as detected by ELISA.    
     
     
         42 . The therapeutic cell composition of  claim 41  comprising about 50% postpartum-derived cells.  
     
     
         43 . The therapeutic cell composition of  claim 42  comprising substantially only postpartum-derived cells.  
     
     
         44 . The therapeutic cell composition of  claim 43  comprising a substantially homogeneous population of postpartum-derived cells.  
     
     
         45 . The therapeutic cell composition of  claim 44  comprising a clonal cell line of postpartum-derived cells.  
     
     
         46 . The therapeutic cell composition of  claim 44  wherein the substantially homogeneous population is umbilical-derived cells.  
     
     
         47 . The therapeutic cell composition of  claim 44  wherein the substantially homogeneous population is substantially free of maternal cells.  
     
     
         48 . The therapeutic cell composition of  claim 44  wherein the substantially homogeneous population is placental-derived cells.  
     
     
         49 . The therapeutic cell composition of  claim 44  wherein the substantially homogeneous population is of neonatal origin.  
     
     
         50 . The therapeutic cell composition of  claim 44  wherein the substantially homogeneous population is of maternal origin.  
     
     
         51 - 58 . (canceled)  
     
     
         59 . A coculture of human postpartum cells and another mammalian cell, wherein at least one of the cells is a cardiomyoblast, angioblast, or hemangioblast or induced to differentiate along a pathway leading to a cardiomyoblast, angioblast or hemangioblast.  
     
     
         60 - 61 . (canceled)