| abstract |
An ex vivo method to induce the differentiation of cells that can be obtained from human cord blood tissue substantially free of blood, where the cells are capable of self-renewal and differentiation, and where the expression by said cells of genes encoding interleukin 8 ; crosslink 1; chemokine ligand 1 (motif C-X-C); chemokine ligand 3 (motif C-X-C); chemokine ligand 6 (CXC motif) and protein 3 induced by tumor necrosis factor alpha is increased in relation to a human cell that is a fibroblast, a mesenchymal stem cell or an iliac crest bone stem cell as characterized in a set of genes, to a chondrogenic phenotype comprising exposing said cells to one or more chondrogenic differentiation inducing agents wherein said cells further comprise the following characteristics: (i) potential to experience at least 40 duplications in the culture; (ii) binding and expansion in a coated or uncoated tissue culture vessel, wherein the coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyiorithin, vitronectin or fibronectin; (iii) production of vimentin and actin of smooth alpha muscle; (iv) production of each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha and HLA-A, B, C, as detected by flow cytometry; (v). lack of expression of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G and HLADR, DP, DQ as detected by flow cytometry; (vi) secretion of MCP-1, IL-6, GCP-2, IL-8, TIMP1, TPO, KGF, HGF, FGF, HBEGF and BDNF as detected by ELISA; (vii) lack of secretion of SDF-1 alpha, VEGF, TGF-beta2, ANG2 and PDGFbb as detected by ELISA; (viii). growth in about 5% to about 20% oxygen (ix). require L-valine for growth; (x) expression of PD-L2 and tissue factor as measured by flow cytometry; (xi) a decrease in the expression of the following genes in relation to a human cell that is a fibroblast, a mesenchymal stem cell, or an iliac crest bone stem cell as characterized in a set of genes: homeotic box 2 of the short stature; 27 kDa heat shock protein 2; chemokine ligand 12 (motif C-X-C) (factor 1 derived from stromal cells); elastin; DKFZp586M2022 cDNA (from clone DKFZp586M2022); homeotic box 2 mesenchyme; homologue 1 of homeotic box of "sine oculis"; crystalline, alpha B; morphogenesis activator 2 associated with "disheveled"; DKFZP586B2420 protein; similar to neuralin 1; tetranectin; domain 3 homology with src (SH3) and rich in cysteines; B lymphocyte translocation gene 1, antiproliferative; cholesterol 25-hydroxylase; transcription factor 3 related to dwarfism; hypothetical protein FLJ23191; interleukin 11 receptor, alpha; procollagen C-endopeptidase enhancer; counterpart 7 of "frizzled"; hypothetical gene BC008967; collagen, type VIII, alpha 1; tenascin C; homeotic box protein iroquois 5; hephaestin; integrin, beta 8; synaptic vesicle glycoprotein 2; CDNA FLJ12280 fis, clone MAMMA1001744; factor 1 similar to the cytokine receptor; calcium-activated potassium channel of intermediate / low conductance, subfamily N, member 4; integrin, alpha 7; DKFZP586L151 protein; co-activator of transcription on the occasion of PDZ binding (TAZ); homologue 2 of homeotic box of "sine oculis"; KIAA1034 protein; early growth response 3; homeotic box of "distal less" 5; hypothetical protein FLJ20373; family 1 of aldo-keto reductases, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); biglican; fibronectin 1; proencephalin; integrin, type beta 1 (with repeat domains similar to EGF); cDNA clone EUROIMAGE 1968422; EphA3; KIAA0367 protein; natriuretic peptide receptor C / guanylate cyclase C (atrial natriuretic peptide receptor C); hypothetical protein FLJ14054; DKFZp564B222 cDNA (from clone DKFZp564B222); membrane protein associated with vesicle 5; fibulin-like extracellular matrix protein 1 containing EGF; similar to protein 3 interaction with BCL2 / adenovirus E1B of 19 kDa; AE binding protein 1; polypeptide 1 of subunit VIIa of cytochrome c oxidase (muscle); neuroblastoma, suppression of tumorigenicity 1; and insulinoid growth factor binding protein 2, 36 kDa; and (xii) production of GCP-2 and NOGO-A as tested by immunocytochemistry. |