Postpartum cells derived from umbilical cord tissue, and methods of making and using the same
First Claim
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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.
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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.
211 Citations
118 Claims
- 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.
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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.
- View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32)
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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. - View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41, 42, 43)
- short stature homeobox 2;
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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 naï
- ve CD4+ T cells, and expresses PD-L2, but not HLA-G, CD178, HLA-DR, HLA-DP, HLA-DQ, CD80, CD86, or B7-H2.
- View Dependent Claims (45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 57, 100, 101, 102)
- 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.
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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:
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(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. - View Dependent Claims (61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 92, 93)
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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. - View Dependent Claims (104)
- the cell capable of growth in an atmosphere containing oxygen from about 5% to at least about 20%;
- 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.
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108. A method for the preparation of therapeutic cells or cultures comprising:
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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. - View Dependent Claims (109, 110, 111)
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- 112. A serum-free medium for the expansion of postpartum-derived cells wherein the medium has one or more growth factors added.
- 116. A therapeutic culture comprising postpartum-derived cells expanded in serum-free medium.
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118. A cell culture bank comprising the cells of 117.
Specification