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 - 19 . (canceled)  
     
     
         20 . A cell produced by a method comprising exposing a postpartum-derived cell (PPDC) to one or more osteogenic differentiation-inducing agents, 
 wherein said PPDC is derived from human postpartum tissue substantially free of blood, wherein said PPDC 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 PPDC requires L-valine for growth; wherein said PPDC is capable of growth in about 5% to about 20% oxygen; wherein said PPDC 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 charnel, 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.    
     
     
         21 - 28 . (canceled)  
     
     
         29 . A cell population comprising the cell of  claim 20 .  
     
     
         30 . The cell population of  claim 29  wherein said cell population is substantially homogeneous.  
     
     
         31 . The cell population of  claim 29  wherein said cell population is heterogeneous.  
     
     
         32 . The cell population of  claim 31  further comprising at least one cell type of bone marrow cells, osteocytes, osteoblasts, osteoclasts, bone lining cells, stem cells or other pluripotent or multipotent cell.  
     
     
         33 - 36 . (canceled)  
     
     
         37 . A cell lysate prepared from the cell population of  claim 29 .  
     
     
         38 . A soluble cell fraction prepared from the cell lysate of  claim 37 .  
     
     
         39 - 40 . (canceled)  
     
     
         41 . An extracellular matrix of the cell population of  claim 29 .  
     
     
         42 - 43 . (canceled)  
     
     
         44 . A composition comprising the cell population of  claim 29  and one or more bioactive factors.  
     
     
         45 - 48 . (canceled)  
     
     
         49 . A pharmaceutical composition comprising a cell of  claim 20  and a pharmaceutically acceptable carrier.  
     
     
         50 - 51 . (canceled)  
     
     
         52 . A pharmaceutical composition comprising the extracellular matrix of  claim 41  and a pharmaceutically acceptable carrier.  
     
     
         53 - 54 . (canceled)  
     
     
         55 . A pharmaceutical composition comprising the lysate of  claim 37  and a pharmaceutically acceptable carrier.  
     
     
         56 - 63 . (canceled)  
     
     
         64 . A matrix comprising a cell population of  claim 29 .  
     
     
         65 - 66 . (canceled)  
     
     
         67 . The matrix of  claim 64  wherein said matrix comprises a three-dimensional scaffold.  
     
     
         68 - 98 . (canceled)  
     
     
         99 . The pharmaceutical composition of  claim 49  wherein said composition comprises an effective amount of said cells to treat a bone or cartilage condition.  
     
     
         100 - 101 . (canceled)  
     
     
         102 . The pharmaceutical composition of  claim 52  wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.  
     
     
         103 - 104 . (canceled)  
     
     
         105 . The pharmaceutical composition of  claim 55  wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.  
     
     
         106 - 107 . (canceled)  
     
     
         108 . The pharmaceutical composition of  claim 49  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.