Cartilage and bone repair and regeneration using postpartum-derived cells

Abstract

Cells derived from postpartum tissue and methods for their isolation and induction to differentiate to cells of a chondrogenic or osteogenic phenotype are provided by the invention. The invention further provides cultures and compositions of the postpartum-derived cells and products related thereto. The postpartum-derived cells of the invention and products related thereto have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications, for example, in the treatment of bone and cartilage conditions.

1 . A postpartum-derived cell comprising a cell derived from human postpartum tissue substantially free of blood, wherein said cell is capable of self-renewal and expansion in culture and has the potential to differentiate into a cell of an osteogenic or chondrogenic phenotype; wherein said cell requires L-valine for growth; wherein said cell is capable of growth in about 5% to about 20% oxygen; wherein said cell further comprises at least one of the following characteristics: 
 (a) production of at least one of granulocyte chemotactic protein 2 (GCP-2), reticulon 1, tissue factor, vimentin, and alpha-smooth muscle actin;    (b) lack of production of at least one of GRO-alpha or oxidized low density lipoprotein receptor, as detected by flow cytometry;    (c) production of at least one of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A,B,C;    (d) 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;    (e) expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is increased for 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 or expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is increased for at least one of C-type lectin superfamily member A2, Wilms tumor 1, aldehyde dehydrogenase 1 family member A2, renin, oxidized low density lipoprotein receptor 1, protein kinase C zeta, clone IMAGE:4179671, hypothetical protein DKFZp564F013, downregulated in ovarian cancer 1, and clone DKFZp547K1113;    (f) 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; cDNA DKFZp586M2022 (from clone DKFZp586M2022); mesenchyme homeobox 2; sine oculis homeobox homolog 1; crystallin, alpha B; dishevelled associated activator of morphogenesis 2; DKFZP586B2420 protein; similar to neuralin 1; tetranectin; 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; hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C; iroquois homeobox protein 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2; 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; 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); cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C); hypothetical protein FLJ14054; cDNA DKFZp564B222 (from clone DKFZp564B222); vesicle-associated membrane protein 5; 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;    (g) secretion of at least one of monocyte chemotactic protein-1, interleukin(IL)-6, IL-8, granulocyte chemotactic protein-2, hepatocyte growth factor, keratinocyte growth factor, fibroblast growth factor, heparin binding-epidermal growth factor, brain derived neurotrophic factor, thrombopoietin, macrophage inflammatory protein (MIP)-1a, RANTES, and tissue inhibitor of matrix metalloprotease 1;    (h) lack of secretion of at least one of transforming growth factor-beta2, angiopoetin-2, platelet derived growth factor-bb, MIP1b, I309, macrophage-derived chemokine, and vascular endothelial growth factor, as detected by ELISA; and    (i) the ability to undergo at least 40 population doublings in culture.    
     
     
         2 . The cell of  claim 1  which has been isolated from a post-partum placenta or fragment thereof by enzymatic dissociation with at least one of a matrix metalloprotease, a neutral protease, and a mucolytic enzyme that digests hyaluronic acid.  
     
     
         3 - 20 . (canceled)  
     
     
         21 . A cell population comprising the postpartum-derived cell of  claim 1 .  
     
     
         22 . The cell population of  claim 21  wherein said cell population is substantially homogeneous.  
     
     
         23 . The cell population of  claim 21  wherein said cell population is heterogeneous.  
     
     
         24 . The cell population of  claim 23  further comprising at least one cell type of bone marrow cells, chondrocytes, chondroblasts, chondrocyte progenitor cells, or stem cells.  
     
     
         25 . A cell population comprising the postpartum-derived cell of claim  12 .  
     
     
         26 . The cell population of  claim 25  wherein said cell population is substantially homogeneous.  
     
     
         27 . The cell population of  claim 25  wherein said cell population is heterogeneous.  
     
     
         28 . The cell population of  claim 27  further comprising at least one cell type of bone marrow cells, chondrocytes, chondroblasts, chondrocyte progenitor cells, stem cells, or other pluripotent or multipotent cell.  
     
     
         29 - 32 . (canceled)  
     
     
         33 . A cell lysate prepared from the cell population of  claim 21 .  
     
     
         34 . A soluble cell fraction prepared from the cell lysate of  claim 33 .  
     
     
         35 . A cell lysate prepared from the cell population of  claim 25 .  
     
     
         36 . A soluble cell fraction prepared from the cell lysate of  claim 35 .  
     
     
         37 - 38 . (canceled)  
     
     
         39 . An extracellular matrix of the cell population of  claim 21 .  
     
     
         40 . An extracellular matrix of the cell population of  claim 25 .  
     
     
         41 . (canceled)  
     
     
         42 . A composition comprising the cell population of  claim 21  and one or more bioactive factors.  
     
     
         43 . A composition comprising the cell population of  claim 25  and one or more bioactive factors.  
     
     
         44 . (canceled)  
     
     
         45 . The composition of  claim 42  wherein said bioactive factor is a chondrogenic differentiation-inducing factor.  
     
     
         46 . The composition of  claim 42  wherein said bioactive factor is an osteogenic differentiation-inducing factor.  
     
     
         47 . A pharmaceutical composition comprising a cell of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         48 - 49 . (canceled)  
     
     
         50 . A pharmaceutical composition comprising the extracellular matrix of  claim 39  and a pharmaceutically acceptable carrier.  
     
     
         51 . A pharmaceutical composition comprising the extracellular matrix of  claim 40  and a pharmaceutically acceptable carrier.  
     
     
         52 . (canceled)  
     
     
         53 . A pharmaceutical composition comprising the lysate of  claim 33  and a pharmaceutically acceptable carrier.  
     
     
         54 . A pharmaceutical composition comprising the lysate of  claim 35  and a pharmaceutically acceptable carrier.  
     
     
         55 . (canceled)  
     
     
         56 . A cell culture comprising at least one cell of  claim 1  in a culture medium.  
     
     
         57 . The cell culture of  claim 56  wherein said culture medium comprises chondrogenic medium or osteogenic medium.  
     
     
         58 . The cell culture of  claim 56  further comprising at least one chondrogenic differentiation-inducing agent.  
     
     
         59 . The cell culture of  claim 58  wherein said chondrogenic differentiation-inducing agent is at least one of transforming growth factor-beta3 or growth and differentiation factor-5.  
     
     
         60 . The cell culture of  claim 56  further comprising at least one osteogenic differentiation-inducing agent.  
     
     
         61 . The cell culture of  claim 60  wherein said osteogenic differentiation-inducing agent is at least one of transforming growth factor-beta 1, bone morphogenic protein (BMP)-2, or BMP4.  
     
     
         62 . A matrix comprising a cell population of  claim 21 .  
     
     
         63 . A matrix comprising a cell population of  claim 25 .  
     
     
         64 . (canceled)  
     
     
         65 . The matrix of  claim 62  wherein said matrix comprises a three-dimensional scaffold.  
     
     
         66 . The matrix of  claim 63  wherein said matrix comprises a three-dimensional scaffold.  
     
     
         67 - 89 . (canceled)  
     
     
         90 . A kit comprising at least one cell of  claim 1  and at least one additional component of a matrix, a hydrating agent, a cell culture substrate, a differentiation-inducing agent, and cell culture media.  
     
     
         91 . The kit of  claim 90  wherein said matrix is a three-dimensional scaffold.  
     
     
         92 . The kit of  claim 91  wherein said cell is seeded on said scaffold.  
     
     
         93 . The kit of  claim 90  wherein said differentiation-inducing agent is an osteogenic differentiation-inducing agent or a chondrogenic differentiation-inducing agent.  
     
     
         94 - 96 . (canceled)  
     
     
         97 . The pharmaceutical composition of  claim 47  wherein said composition comprises an effective amount of said cells to treat a bone or cartilage condition.  
     
     
         98 - 99 . (canceled)  
     
     
         100 . The pharmaceutical composition of  claim 50  wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.  
     
     
         101 . The pharmaceutical composition of  claim 51  wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.  
     
     
         102 . (canceled)  
     
     
         103 . The pharmaceutical composition of  claim 53  wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.  
     
     
         104 . The pharmaceutical composition of  claim 54  wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.  
     
     
         105 . (canceled)  
     
     
         106 . The pharmaceutical composition of  claim 47  further comprising at least one other cell type of stem cells, bone marrow cells, chondrocytes, chondroblasts, osteocytes, osteoblasts, osteoclasts, bone lining cells, and other bone or cartilage progenitor cells.  
     
     
         107 - 108 . (canceled)