US5032508A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Three-dimensional cell and tissue culture system
Assignee: REGENEMED Inc A CALIFORNIA Corp
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Filing: 1989-09-01
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Granted: 1991-07-16
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Expires: 2009-09-01
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 02:42
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0062; A; A61; A61L; A61L27/00; A61L27/36; A61L27/38; A61L27/50; A61L27/60; C12N5/06; C12N5/0602; C12N5/0618; C12N5/0652; C12N5/0669; C12N5/067; C12N5/0671; C12N5/0676; C12N5/0677; C12N5/0697; C12N5/0698; A61K; A61K35/00; A61K35/12; A61K2035/124; C12N2502/00; C12N2502/08; C12N2502/081; C12N2502/086; C12N2502/09; C12N2502/091; C12N2502/094; C12N2502/097; C12N2502/13; C12N2502/1305; C12N2502/1323; C12N2502/28; C12N2502/30; C12N2503/00; C12N2503/04; C12N2510/00; C12N2533/00; C12N2533/30; C12N2533/40; C12N2533/70
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
101 words
A method for testing the cytological effect of a test substance comprising: (a) exposing a three dimensional cell culture to the test substance, in which the three dimensional cell culture comprises parenchymal cells grown 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 bridges by the stromal cells; and (b) determining the effect of the test substance by measuring a change in the three dimensional cell culture.
Claim 2
Independent Claim
101 words
A method for testing the cytological effect of a test substance comprising: (a) exposing a three dimensional cell culture to the test substance, in which the three dimensional cell culture comprises parenchymal cells grown 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 bridges by the stromal cells; and (b) determining the effect of the test substance by measuring a change in the three dimensional cell culture.
Claim 3
Dependent Claim
54 words
The method of claim 1 for testing the cytotoxicity of a test substance, wherein the effect of the test substance is determined by measuring the number of damaged or deed cells in the three dimensional cell culture, in which the number of damaged or dead cells correlates with the cytotoxicity of the test substance.
Claim 4
Dependent Claim
54 words
The method of claim 1 for testing the growth/regulatory activity of a test substance, wherein the effect of the test substance is determined by analyzing the cellular content of the three dimensional cell culture, in which an increase in cell number or a cell type correlates with the growth/regulatory activity of the test substance.
Claim 5
Dependent Claim
54 words
The method of claim 1 for testing the metabolic activity of a test substance, wherein the effect of the test substance is determined by measuring a metabolite produced by the three dimensional cell culture, in which an increase or decrease in the amount of metabolite produced correlates with the activity of the test substance.
Claim 6
Dependent Claim
60 words
The method of claim 1 for testing the allergenic effect of a test substance, wherein the three dimensional cell cultures containing mast cells, and the effect of the test substance is determined by measuring the release of a vasoactive mediator by the mast cells, in which release of the vasoactive mediator correlates with the allergenic effect of the test substance.
Claim 7
Dependent Claim
22 words
The method for testing the cytological effect of a test substance according to claim 1 in which the stromal cells are fibroblasts.
Claim 8
Dependent Claim
38 words
The method for testing the cytological effect of a test substance according to claim 1 in which the stromal cells are a combination of fibroblasts and endothelial cells, perictyes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 9
Dependent Claim
25 words
The method for testing the cytological effect of a test substance according to claim 1 in which the framework is composed of a biodegradable material.
Claim 10
Dependent Claim
31 words
The method for testing the cytological effect of a test substance according to claim 8 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 11
Dependent Claim
25 words
The method for testing the cytological effect of a test substance according to claim 1 in which the framework is composed of a non-biodegradable material.
Claim 12
Dependent Claim
41 words
The method for testing the cytological effect of a test substance according to claim 10 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 13
Dependent Claim
27 words
The method for testing the cytological effect of a test substance according to claim 8, 9, 10 or 11 in which the framework is pre-coated with collagen.
Claim 14
Dependent Claim
33 words
The method for testing the cytological effect of a test substance according to claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 in which the framework is a mesh.
Claim 15
Dependent Claim
22 words
The method for testing the cytological effect of a test substance according to claim 12 in which the framework is a mesh.
Claim 16
Dependent Claim
23 words
The method for testing the cytological effect of a test substance according to claim 1 in which the parenchymal cells are hematopoietic cells.
Claim 17
Dependent Claim
24 words
The method for testing the cytological effect of a test substance according to claim 1 in which the parenchymal cells are melanocytes and keratinocytes.
Claim 18
Dependent Claim
22 words
The method for testing the cytological effect of a test substance according to claim 1 in which the parenchymal cells are hepatocytes.
Claim 19
Dependent Claim
24 words
The method for testing the cytological effect of a test substance according to claim 11 in which the parenchymal cells are pancreatic acinar cells.
Claim 20
Dependent Claim
23 words
The method for testing the cytological effect of a test substance according to claim 1 in which the parenchymal cells are kidney cells.
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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) | 1994-07-15 | 1995-01-14 | 1995-07-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-15 | 1999-01-14 | 1999-07-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-15 | 2003-01-14 | 2003-07-16 | $7700.00 | Window Closed / Lapsed |