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US5266480A
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Three-dimensional skin culture system
Assignee: Advanced BioHealing Inc
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Filing: 1990-08-30
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Granted: 1993-11-30
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Expires: 2010-08-30
Abstract
Abstract The present invention relates to a three-dimensional cell culture system which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. In accordance with the invention, cells derived from a desired tissue are inoculated and grown on a pre-established stromal support matrix. The stromal support matrix comprises stromal cells, such as fibroblasts actively growing on a three-dimensional matrix. Stromal cells may also include other cells found in loose connective tissue such as endothelial cells, macrophages/monocytes, adipocytes, pericytes, reticular cells found in bone marrow stroma, etc. The stromal matrix provides the support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts found in vivo.
Inventors
- 👤 Gail K. Naughton
- 👤 Brian A. Naughton
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:33
CPC Classifications
A; A61; A61L; A61L27/00; A61L27/36; A61L27/3604; A61B; A61B90/00; A61K; A61K35/00; A61K35/12; A61K35/28; A61L27/38; A61L27/3886; A61L27/50; A61L27/60; C; C12; C12N; C12N5/00; C12N5/06; C12N5/0697; C12N5/0698; A61B10/00; A61B10/02; A61B10/0233; A61B10/025; A61B2010/0258; A61K2035/124; C12N2500/00; C12N2500/30; C12N2500/34; C12N2501/00; C12N2501/10; C12N2501/11; C12N2501/30; C12N2501/33; C12N2501/335; C12N2501/38; C12N2501/39; C12N2502/00; C12N2502/09; C12N2502/091; C12N2502/094; C12N2502/13; C12N2502/1323; C12N2503/00; C12N2503/04; C12N2510/00; C12N2533/00; C12N2533/30; C12N2533/40; C12N2533/70; Y; Y10; Y10S; Y10S623/00; Y10S623/915
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a living stromal tissue prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three dimensional structure having interstitial spaces bridged by the stromal cells.
Claim 2
Dependent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a living stromal tissue prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three dimensional structure having interstitial spaces bridged by the stromal cells.
Claim 3
Dependent Claim
14 words
The three-dimensional skin culture of claim 1 in which the stromal cells comprise fibroblasts.
Claim 4
Dependent Claim
30 words
The three-dimensional skin culture of claim 1 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 5
Dependent Claim
17 words
The three-dimensional skin culture of claim 1 in which the framework is composed of a biodegradable material.
Claim 6
Dependent Claim
23 words
The three-dimensional skin culture of claim 4 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 7
Dependent Claim
17 words
The three-dimensional skin culture of claim 1 in which the framework is composed of a non-biodegradable material.
Claim 8
Dependent Claim
33 words
The three-dimensional skin culture of claim 6 in which the non-biodegradable material is a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 9
Dependent Claim
19 words
The three-dimensional skin culture of claims 4, 5, 6 or 7 in which the framework is pre-coated with collagen.
Claim 10
Dependent Claim
21 words
The three-dimensional skin culture of claims 1, 2, 3, 4, 5, 6 or 7 in which the framework is a mesh.
Claim 11
Dependent Claim
14 words
The three-dimensional skin culture of claim 9 in which the framework is a mesh.
Claim 12
Dependent Claim
55 words
A three dimensional culture of dermis prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells, in which the stromal cells are confluent.
Claim 13
Dependent Claim
55 words
A three dimensional culture of dermis prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells, in which the stromal cells are confluent.
Claim 14
Dependent Claim
14 words
The three-dimensional dermis culture of claim 11 in which the stromal cells are fibroblasts.
Claim 15
Dependent Claim
30 words
The three-dimensional dermis culture of claim 11 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 16
Dependent Claim
17 words
The three-dimensional dermis culture of claim 11 in which the framework is composed of a biodegradable material.
Claim 17
Dependent Claim
23 words
The three-dimensional dermis culture of claim 14 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 18
Dependent Claim
17 words
The three-dimensional dermis culture of claim 11 in which the framework is composed of a non-biodegradable material.
Claim 19
Dependent Claim
33 words
The three-dimensional dermis culture of claim 16 in which the non-biodegradable material is a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 20
Dependent Claim
19 words
The three-dimensional dermis culture of claims 14, 15, 16 or 17 in which the framework is pre-coated with collagen.
Claim 21
Dependent Claim
21 words
The three-dimensional dermis culture of claims 11, 12, 13, 14, 15, 16 or 17 in which the framework is a mesh.
Claim 22
Dependent Claim
14 words
The three-dimensional dermis culture of claim 18 in which the framework is a mesh.
Claim 23
Independent Claim
82 words
A method for culturing skin cells in vitro comprising: (a) inoculating melanocytes and keratinocytes onto a living stromal tissue prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three dimensional structure having interstitial spaced bridges by the stromal cells; and (b) incubating the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 24
Independent Claim
82 words
A method for culturing skin cells in vitro comprising: (a) inoculating melanocytes and keratinocytes onto a living stromal tissue prepared in vitro, comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three dimensional structure having interstitial spaced bridges by the stromal cells; and (b) incubating the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 25
Dependent Claim
13 words
The method according to claim 21 in which the stromal cells are fibroblasts.
Claim 26
Dependent Claim
29 words
The method according to claim 21 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 27
Dependent Claim
16 words
The method according to claim 21 in which the framework is composed of a biodegradable material.
Claim 28
Dependent Claim
22 words
The method according to claim 24 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 29
Dependent Claim
16 words
The method according to claim 21 in which the framework is composed of a non-biodegradable material.
Claim 30
Dependent Claim
32 words
The method according to claim 26 in which the non-biodegradable material is a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 31
Independent Claim
18 words
The method according to claims 24, 25, 26 or 27 in which the framework is pre-coated with collagen.
Claim 32
Independent Claim
20 words
The method according to claims 21, 22, 23, 24, 25, 26 or 27 in which the framework is a mesh.
Claim 33
Dependent Claim
13 words
The method according to claim 29 in which the framework is a mesh.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-11-29 | 1997-05-31 | 1997-11-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-11-29 | 2001-05-31 | 2001-11-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-11-29 | 2005-05-31 | 2005-11-30 | $7700.00 | Window Closed / Lapsed |