IMPLANTABLE COMPOSITIONS FOR REPAIRING OSTEOCHONDRAL DEFECTS
Compositions are described that are suitable for implantation in a human and that contain a scaffold housing cells having an osteocytic phenotype and, attached to the scaffold, cartilage material containing a population of chondrocytes.
1 . A composition comprising a collagen coated three-dimensional scaffold housing a population of osteoblasts and, attached thereto, cartilage material comprising a population of chondrocytes.
2 . The composition of claim 1 , wherein said population of chondrocytes is isogenic.
3 . The composition of claim 1 , wherein said population of osteoblasts is isogenic.
4 . The composition of claim 1 , wherein said population of chondrocytes is a clonal population.
5 . The composition of claim 1 , wherein said population of osteoblasts is a clonal population.
6 . The composition of claim 1 , wherein said composition is suitable for implantation in a human.
7 . The composition of claim 1 , wherein said collagen coated three-dimensional scaffold is composed of tri-calcium phosphate, alumina, titania, poly-lactic acid, poly-1-glycolic acid, polyetheretherketone (PEEK), polycaprolactone, or collagen.
8 . The composition of claim 1 , wherein said collagen coated three-dimensional scaffold is biodegradable.
9 . The composition of claim 1 , further comprising a chondrocyte differentiation medium.
10 . The composition of claim 1 , wherein said composition is housed within a sterile container.
11 . The composition of claim 1 , said composition further comprising a cryopreservative.
12 . The composition of claim 11 , wherein said cryopreservative is dimethylsulfoxide (DMSO).
13 . The composition of claim 12 , wherein said cryopreservative is 1 to 10% DMSO.
14 . The composition of claim 11 , wherein said cryopreservative is fetal bovine serum, human serum, or human serum albumin in combination with one or more of the following: DMSO, trehalose, and dextran.
15 . The composition of claim 14 , wherein said cryopreservative is human serum, DMSO, and trehalose.
16 . The composition of claim 14 , wherein said cryopreservative is fetal bovine serum and DMSO.
17 . An article of manufacture comprising a purified population of human fetal blood multi-lineage progenitor cells (MLPC) or a clonal line of human fetal blood MLPC, a differentiation medium for inducing differentiation of said MLPC into cells having an osteoblast cell phenotype, and a differentiation medium for inducing differentiation of said MLPC into cells having a chondrogenic cell phenotype, wherein said MLPC are positive for CD9, negative for CD45, negative for CD34, and negative for SSEA-4.
18 . The article of manufacture of claim 17 further comprising a growth substrate coated with collagen.
19 . The article of manufacture of claim 18 , wherein said growth substrate is a collagen-coated culturing device.
20 . The article of manufacture of claim 17 further comprising a collagen coated three-dimensional scaffold.
21 . The article of manufacture of claim 20 , wherein said collagen coated three-dimensional scaffold is composed of tri-calcium phosphate, alumina, titania, poly-lactic acid, poly-1-glycolic acid, polyetheretherketone (PEEK), poly caprolactone, or collagen.
22 . The article of manufacture of claim 17 , wherein said MLPC are further positive for CD13, CD29, CD44, CD73, CD90 and CD105, and further negative for CD10, CD41, Stro-1, and SSEA-3.
23 . The article of manufacture of claim 17 , wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.
24 . The article of manufacture of claim 17 , wherein said osteoblast differentiation medium comprises dexamethasone, L-glutamine, ascorbate, andβ-glycerophosphate.
25 . The article of manufacture of claim 17 , wherein said chondrogenic differentiation medium comprises ascorbate, dexamethasone and transforming growth factor beta 3.
26 . A method of producing an implant for repairing osteochondral defects, said method comprising contacting a collagen coated three-dimensional scaffold housing a population of osteoblasts with cartilage material comprising a population of chondrocytes in the presence of a chondrocyte differentiation medium to produce said implant; and culturing said implant for a time sufficient for said cartilage material to securely attach to said collagen-coated scaffold.
27 . The method of claim 26 , wherein said population of chondrocytes is isogenic.
28 . The method of claim 26 , wherein said population of osteoblasts is isogenic.
29 . The method of claim 26 , wherein said population of chondrocytes is a clonal population.
30 . The method of claim 26 , wherein said population of osteoblasts is a clonal population.
31 . The method of claim 30 , wherein said clonal population of osteoblasts is obtained by culturing a clonal line of MLPC with a differentiation medium for inducing differentiation of said MLPC into cells having an osteoblast cell phenotype, wherein said MLPC are positive for CD9, negative for CD45, negative for CD34, and negative for SSEA-4 before differentiation.
32 . The method of claim 31 , wherein said MLPC are further positive for CD13, CD29, CD44, CD73, CD90 and CD105, and further negative for CD10, CD41, Stro-1, and SSEA-3.
33 . The method of claim 31 , wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.
34 . The method of claim 29 , wherein said clonal population of chondrocytes is obtained by culturing a clonal line of MLPC on a collagen coated growth substrate with a differentiation medium effective for inducing differentiation of said MLPC into cells having said chondrocyte phenotype, wherein said MLPC are positive for CD9, negative for CD45, negative for CD34, and negative for SSEA-4.
35 . The method of claim 34 , wherein said collagen coated growth substrate is a collagen coated culturing device.
36 . The method of claim 26 , further comprising testing cells having said chondrocyte phenotype for intracellular aggrecan, intracellular collagen type II, intracellular SOX9, or cell surface TGF-β receptor.
37 . The method of claim 26 , further comprising testing cells having said osteoblast phenotype for mineralization or expression of collagen type I, osteocalcin, osteopontin, or alkaline phosphatase.