US5512475A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Three-dimensional skin cell and tissue culture system
Assignee: Shire Regenerative Medicine LLC
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Filing: 1995-04-06
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Granted: 1996-04-30
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Expires: 2015-04-06
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 04:40
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: 41
Claim 1
Independent Claim
80 words
A method for transplantation or implantation of a three-dimensional skin cell culture comprising implanting a culture of melanocyte and keratinocyte cells cultured on a living stromal tissue prepared in vitro, said living stromal tissue 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 so that a tissue equivalent is formed.
Claim 2
Independent Claim
80 words
A method for transplantation or implantation of a three-dimensional skin cell culture comprising implanting a culture of melanocyte and keratinocyte cells cultured on a living stromal tissue prepared in vitro, said living stromal tissue 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 so that a tissue equivalent is formed.
Claim 3
Dependent Claim
13 words
The method according to claim 1 in which the stromal cells are fibroblasts.
Claim 4
Dependent Claim
29 words
The method according to 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
16 words
The method according to claim 1 in which the framework is composed of a biodegradable material.
Claim 6
Dependent Claim
22 words
The method according to claim 4 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 7
Dependent Claim
16 words
The method according to claim 1 in which the framework is composed of a non-biodegradable material.
Claim 8
Dependent Claim
32 words
The method according to 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
Independent Claim
77 words
A method for transplantation or implantation of a three-dimensional culture comprising implanting in vivo melanocytes and keratinocytes cultured on a living stromal tissue prepared in vitro, said living stromal tissue comprising confluent 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 so that a tissue equivalent is formed.
Claim 10
Independent Claim
77 words
A method for transplantation or implantation of a three-dimensional culture comprising implanting in vivo melanocytes and keratinocytes cultured on a living stromal tissue prepared in vitro, said living stromal tissue comprising confluent 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 so that a tissue equivalent is formed.
Claim 11
Dependent Claim
13 words
The method according to claim 8 in which the stromal cells are fibroblasts.
Claim 12
Dependent Claim
29 words
The method according to claim 8 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 13
Dependent Claim
16 words
The method according to claim 8 in which the framework is composed of a biodegradable material.
Claim 14
Dependent Claim
22 words
The method according to claim 11 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 15
Dependent Claim
16 words
The method according to claim 8 in which the framework is composed of a non-biodegradable material.
Claim 16
Dependent Claim
32 words
The method according to claim 13 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 17
Dependent Claim
22 words
The method according to claim 4, 5, 11, 12, 6, 7, 13 or 14 in which the framework is pre-coated with collagen.
Claim 18
Dependent Claim
13 words
The method according to claim 15 in which the framework is a mesh.
Claim 19
Dependent Claim
27 words
The method according to claim 1, 8, 2, 3, 9, 10, 4, 5, 11, 12, 6, 7, 13 or 14 in which the framework is a mesh.
Claim 20
Independent Claim
107 words
A method for determining the effect of a drug on melanocyte and keratinocyte cells in culture comprising: (a) exposing a three-dimensional skin cell culture to the drug, in which the three-dimensional skin cell culture comprises melanocyte and keratinocyte cells grown on a living stromal tissue prepared in vitro, said living stromal tissue 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; and (b) determining the effect of the drug on the melanocyte and keratinocyte cells in culture.
Claim 21
Independent Claim
107 words
A method for determining the effect of a drug on melanocyte and keratinocyte cells in culture comprising: (a) exposing a three-dimensional skin cell culture to the drug, in which the three-dimensional skin cell culture comprises melanocyte and keratinocyte cells grown on a living stromal tissue prepared in vitro, said living stromal tissue 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; and (b) determining the effect of the drug on the melanocyte and keratinocyte cells in culture.
Claim 22
Dependent Claim
20 words
The method for determining the effect of a drug according to claim 18 in which the stromal cells are fibroblasts.
Claim 23
Dependent Claim
36 words
The method for determining the effect of a drug according to claim 18 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 24
Dependent Claim
23 words
The method for determining the effect of a drug according to claim 18 in which the framework is composed of a biodegradable material.
Claim 25
Dependent Claim
29 words
The method for determining the effect of a drug according to claim 21 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 26
Dependent Claim
23 words
The method for determining the effect of a drug according to claim 18 in which the framework is composed of a non-biodegradable material.
Claim 27
Dependent Claim
39 words
The method for determining the effect of a drug according to claim 23 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 28
Dependent Claim
25 words
The method for determining the effect of a drug according to claim 21, 22, 23 or 24 in which the framework is pre-coated with collagen.
Claim 29
Dependent Claim
13 words
The method according to claim 25 in which the framework is a mesh.
Claim 30
Dependent Claim
20 words
The method according to claim 18, 19, 20, 21, 22, 23 or 24 in which the framework is a mesh.
Claim 31
Independent Claim
128 words
A method for studying the mechanisms of a disease or condition in a patient, which disease or condition has a detectable affect on melanocyte and keratinocyte cells in culture, comprising: (a) inoculating melanocyte and keratinocyte cells obtained from the patient onto a living stromal tissue prepared in vitro, said living stromal tissue 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; (b) incubating the inoculated living stromal tissue in a nutrient medium so that the inoculated melanocyte and keratinocyte cells proliferate in culture; and (c) analyzing the proliferated cells in culture for markers of the disease or condition.
Claim 32
Independent Claim
128 words
A method for studying the mechanisms of a disease or condition in a patient, which disease or condition has a detectable affect on melanocyte and keratinocyte cells in culture, comprising: (a) inoculating melanocyte and keratinocyte cells obtained from the patient onto a living stromal tissue prepared in vitro, said living stromal tissue 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; (b) incubating the inoculated living stromal tissue in a nutrient medium so that the inoculated melanocyte and keratinocyte cells proliferate in culture; and (c) analyzing the proliferated cells in culture for markers of the disease or condition.
Claim 33
Dependent Claim
13 words
The method according to claim 28 in which the stromal cells are fibroblasts.
Claim 34
Dependent Claim
29 words
The method according to claim 28 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 35
Dependent Claim
16 words
The method according to claim 28 in which the framework is composed of a biodegradable material.
Claim 36
Dependent Claim
22 words
The method according to claim 31 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 37
Dependent Claim
16 words
The method according to claim 28 in which the framework is composed of a non-biodegradable material.
Claim 38
Dependent Claim
32 words
The method according to claim 33 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 39
Dependent Claim
18 words
The method according to claim 31, 32, 33 or 34 in which the framework is pre-coated with collagen.
Claim 40
Dependent Claim
13 words
The method according to claim 35 in which the framework is a mesh.
Claim 41
Dependent Claim
20 words
The method according to claim 28, 29, 30, 31, 32, 33 or 34 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) | 1999-04-30 | 1999-10-30 | 2000-04-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-04-30 | 2003-10-30 | 2004-04-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-04-30 | 2007-10-30 | 2008-04-30 | $7700.00 | Window Closed / Lapsed |