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US 2003/0211602 A1Published application
Isolation of mesenchymal stem cells and use thereof
OwnerAnthony Atalainventor
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Abstract
The present invention provides a method of isolating mesenchymal stem cells from postnatal human tissue such as postnatal human foreskin tissue. The present invention additionally provides a method of differentiating the isolated mesenchymal stem cells into various lineages, such as osteogenic, adipogenic, and chondrogenic lineages.
Claims9
A method for obtaining human mesenchymal stem cells, the method comprising:
- (a) providing a specimen of human connective tissue;
- (b) holding the specimen in a sterile environment for at least 2 days at about 4C;
- (c) isolating a dermal layer from the specimen;
- (d) contacting the dermal layer with collagenase to provide a collagenase cell suspension;
- (e) collecting supernatant from the collagenase cell suspension;
- (f) centrifuging the supernatant to obtain a cell product;
- (g) culturing the cell product under conditions to promote cell growth;
- and
- (h) isolating mesenchymal stem cells from the culture.
6 more elements
A method for obtaining osteoblasts, the method comprising:
- (a) providing a specimen of human connective tissue;
- (b) holding the specimen in a sterile environment for at least 2 days at about 4 C.;
- (c) isolating a dermal layer from the tissue;
- (d) contacting the dermal layer with collagenase to provide a collagenase cell suspension;
- (e) collecting supernatant from the collagenase cell suspension;
- (f) centrifuging the supernatant to obtain a cell product;
- and (g) culturing the cell product under conditions to promote cell growth in contact with dexamethasone, beta-glycerophosphate and ascorbic acid-2-phosphate in amounts sufficient to induce human mesenchymal stem cells to undergo osteogenic differentiation.
4 more elements
A method for obtaining adipocytes, the method comprising:
- (a) providing a specimen of human connective tissue;
- (b) holding the specimen in a sterile environment for at least 2 days at about 4 C.;
- (c) isolating a dermal layer from the tissue;
- (d) contacting the dermal layer with collagenase to provide a collagenase cell suspension;
- (e) collecting supernatant from the collagenase cell suspension;
- (f) centrifuging the supernatant to obtain a cell product;
- and
- (g) culturing the cell product under conditions to promote cell growth in contact with dexamethasone, 3-isobutyl-1-methylxanthine, insulin, and indomethacin in an amount sufficient to induce human mesenchymal stem cells to undergo adipogenic differentiation.
5 more elements
A method for obtaining chondrocytes, the method comprising:
- (a) providing a specimen of human connective tissue;
- (b) holding the specimen in a sterile environment for at least 2 days at about 4 C.;
- (c) isolating a dermal layer from the tissue;
- (d) contacting the dermal layer with collagenase to provide a collagenase cell suspension;
- (e) collecting supernatant from the collagenase cell suspension;
- (f) centrifuging the supernatant to obtain a cell product;
- and
- (g) culturing the cell product under conditions to promote cell growth in contact with in contact with L-glutamine, MEM nonessential amino acids, 2-mercaptoethanol, dexamethasone, ascorbic acid-2-phosphate, and transforming growth factor 3 in an amount sufficient to induce human mesenchymal stem cells to undergo chondrogenic differentiation.
5 more elements
Dependencies are inferred from the claim text.