Use of adipose tissue-derived stromal cells for chondrocyte differentiation and cartilage repair
First Claim
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1. A composition comprising:
- (a) an isolated human adipose tissue derived stromal cell; and
(b) a chemically defined culture medium having (i) a chondroinductive agent capable of activating any cellular transduction pathway leading to the mature chondrocyte phenotype (ii) an antibiotic (iii) a nutrient supplement (iv) ascorbate or related vitamin C analogue and (v) a glucocorticoid or other chemical agent capable of activating the cellular glucocorticoid receptor.
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Abstract
Methods and compositions for directing adipose-derived stromal cells cultivated in vitro to differentiate into cells of the chondrocyte lineage are disclosed. The invention further provides a variety of chondroinductive agents which can be used singly or in combination with other nutrient components to induce chondrogenesis in adipose-derived stromal cells either in cultivating monolayers or in a biocompatible lattice or matrix in a three-dimensional configuration. Use of the differentiated chondrocytes for the therapeutic treatment of a number of human conditions and diseases including repair of cartilage in vivo is disclosed.
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Citations
31 Claims
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1. A composition comprising:
- (a) an isolated human adipose tissue derived stromal cell; and
(b) a chemically defined culture medium having (i) a chondroinductive agent capable of activating any cellular transduction pathway leading to the mature chondrocyte phenotype (ii) an antibiotic (iii) a nutrient supplement (iv) ascorbate or related vitamin C analogue and (v) a glucocorticoid or other chemical agent capable of activating the cellular glucocorticoid receptor. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
- (a) an isolated human adipose tissue derived stromal cell; and
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16. A method for differentiating adipose tissue derived stromal cells into chondrocytic cells, comprising:
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a) pelleting said stromal cells and preadipocytes by centrifuging between 50,000 to 5 million cells at 500×
g for 2 to 20 minutes in sterile tubes containing a medium;
b) plating isolated stromal cells and preadipocytes at a density of 500 to 20,000 cells/cm2 in a differentiating medium;
c) supplementing said medium with;
i) a chondroinductive agent capable of activating any cellular signal transduction pathway leading to the mature chondrocyte phenotype ii) an antibiotic iii) a nutrient supplement with 1 to 20% fetal bovine serum or 1 to 20% horse serum or any other biological or synthetic equivalent combination of proteins iv) ascorbate or related vitamin C analog v) a glucocorticoid or other chemical agent capable of activating the cellular glucocorticoid receptor; and
d) incubating said cells at about 31°
C. to 37°
C. for about 3-4 weeks with 5% CO2 and between 1% and 20% oxygen; and
,e) determining the extent of differentiation of the isolated cells.
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17. A method for differentiating adipose tissue derived stromal cells into chondrocytic cells, comprising:
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a) suspending stromal cells and preadipocytes at a concentration of 0.5 to 10 million cells per ml in calcium alginate or any other biocompatible lattice or matrix capable of supporting chondrogenesis in a three-dimensional configuration;
b) transferring cells to 35 mm culture dishes and plating cells at a density of 500 to 20,000 cells/cm2 in a differentiating medium comprising a chemically defined culture medium having or supplemented with;
i) a chondroinductive agent capable of activating any cellular transduction pathway leading to the mature chondrocyte phenotype;
ii) an antibiotic;
iii) a nutrient supplement with 1 to 20% fetal bovine serum or 1 to 20% horse serum or any other biological or synthetic equivalent combination of proteins;
iv) ascorbate or related vitamin C analog;
v) a glucocorticoid or other chemical agent capable of activating the cellular glucocorticoid receptor; and
c) incubating said cells at about 31 to 37°
C. for about 3-4 weeks in an incubator with 5% CO2 and between 1% and 20% oxygen, andd) determining the extent of differentiation of the isolated cells.
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18. A composition comprising:
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a) an isolated adipose tissue derived stromal cell that has been induced to express at least one characteristic of a chondrocyte; and
b) a biocompatible matrix.
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- 19. The composition of claim 19, wherein the matrix is capable of supporting chondrogenesis.
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21. An isolated adipose tissue derived stromal cell that has been induced to express at least one characteristic of a chondrocyte wherein a foreign nucleic acid has been introduced into the cell.
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22. A synthetic cartilage patch comprising an isolated adipose tissue derived stromal cell that has been induced to express at least one characteristic of a chondrocyte.
- 23. The cartilage patch of claim 23, wherein the patch is grown in pre-shaped culture wells to produce a patch of specific dimensions.
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29. The culture wells of claim 29, wherein the pliable or moldable biocompatible material is selected from the group consisting of agarose, glass, untreated cell culture plastic, and polytetrafluoroethylene.
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30. An isolated human adipose tissue derived stromal cell that has been induced to express at least one characteristic of a chondrocyte.
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31. The cell of claim 31, implanted in vivo.
Specification