Patent US5580781A successfully refreshed from Google Patents.
US5580781A
Expired / Lapsed
US
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Three-dimensional tumor cell and tissue culture system
Assignee: REGENEMED Inc A CALIFORNIA Corp
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Filing: 1995-04-06
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Granted: 1996-12-03
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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
2026-11-01 10:17
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: 65
Claim 1
Independent Claim
55 words
A three-dimensional tumor culture, comprising tumor cells 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
55 words
A three-dimensional tumor culture, comprising tumor cells 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 cell culture of claim 1 in which the stromal cells are fibroblasts.
Claim 4
Dependent Claim
30 words
The three-dimensional cell 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 cell culture of claim 1 in which the framework is composed of a biodegradable material.
Claim 6
Dependent Claim
23 words
The three-dimensional cell 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 cell culture of claim 1 in which the framework is composed of a non-biodegradable material.
Claim 8
Dependent Claim
33 words
The three-dimensional cell 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 cell culture of claim 4, 5, 6 or 7 in which the framework is pre-coated with collagen.
Claim 10
Dependent Claim
21 words
The three-dimensional cell culture of claim 1, 2, 3, 4, 5, 6 or 7 in which the framework is a mesh.
Claim 11
Dependent Claim
14 words
The three-dimensional cell culture of claim 8 in which the framework is a mesh.
Claim 12
Independent Claim
60 words
A method for culturing tumor cells in vitro, comprising culturing tumor cells inoculated 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 spaces bridged by the stromal cells.
Claim 13
Independent Claim
60 words
A method for culturing tumor cells in vitro, comprising culturing tumor cells inoculated 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 spaces bridged by the stromal cells.
Claim 14
Dependent Claim
13 words
The method according to claim 11 in which the stromal cells are fibroblasts.
Claim 15
Dependent Claim
29 words
The method according to 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
16 words
The method according to claim 11 in which the framework is composed of a biodegradable material.
Claim 17
Dependent Claim
22 words
The method according to claim 14 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 18
Dependent Claim
16 words
The method according to claim 11 in which the framework is composed of a non-biodegradable material.
Claim 19
Dependent Claim
32 words
The method according to 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
18 words
The method according to claim 14, 15, 16 or 17 in which the framework is pre-coated with collagen.
Claim 21
Dependent Claim
20 words
The method according to claim 11, 12, 13, 14, 15, 16 or 17 in which the framework is a mesh.
Claim 22
Dependent Claim
13 words
The method according to claim 18 in which the framework is a mesh.
Claim 23
Independent Claim
74 words
A method for transplantation or implantation of tumor cells into non-human animals which comprises implanting in vivo the tumor cells 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 so that a tumor is formed.
Claim 24
Independent Claim
74 words
A method for transplantation or implantation of tumor cells into non-human animals which comprises implanting in vivo the tumor cells 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 so that a tumor is formed.
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
Dependent Claim
18 words
The method according to claim 24, 25, 26 or 27 in which the framework is pre-coated with collagen.
Claim 32
Dependent Claim
20 words
The method according to claim 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 28 in which the framework is a mesh.
Claim 34
Independent Claim
90 words
A method for testing the effect of a drug comprising: (a) exposing a three-dimensional tumor cell culture to the drug, in which the three-dimensional cell culture comprises tumor 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 bridged by the stromal cells; and (b) determining the effect of the drug on the cells in culture.
Claim 35
Independent Claim
90 words
A method for testing the effect of a drug comprising: (a) exposing a three-dimensional tumor cell culture to the drug, in which the three-dimensional cell culture comprises tumor 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 bridged by the stromal cells; and (b) determining the effect of the drug on the cells in culture.
Claim 36
Dependent Claim
20 words
The method for testing the effect of a drug according to claim 31 in which the stromal cells are fibroblasts.
Claim 37
Dependent Claim
36 words
The method for testing the effect of a drug according to claim 31 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 38
Dependent Claim
23 words
The method for testing the effect of a drug according to claim 31 in which the framework is composed of a biodegradable material.
Claim 39
Dependent Claim
29 words
The method for testing the effect of a drug according to claim 34 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 40
Dependent Claim
23 words
The method for testing the effect of a drug according to claim 31 in which the framework is composed of a non-biodegradable material.
Claim 41
Dependent Claim
39 words
The method for testing the effect of a drug according to claim 36 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 42
Dependent Claim
25 words
The method for testing the effect of a drug according to claim 34, 35, 36 or 37 in which the framework is pre-coated with collagen.
Claim 43
Dependent Claim
20 words
The method according to claim 31, 32, 33, 34, 35, 36 or 37 in which the framework is a mesh.
Claim 44
Dependent Claim
13 words
The method according to claim 38 in which the framework is a mesh.
Claim 45
Independent Claim
100 words
A method for diagnosing or monitoring a malignancy in a patient, comprising: (a) inoculating tumor cells obtained from the patient 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 spaces bridged by the stromal cells; (b) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (c) determining the presence of malignant cells in the proliferated cells in culture.
Claim 46
Independent Claim
100 words
A method for diagnosing or monitoring a malignancy in a patient, comprising: (a) inoculating tumor cells obtained from the patient 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 spaces bridged by the stromal cells; (b) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (c) determining the presence of malignant cells in the proliferated cells in culture.
Claim 47
Dependent Claim
19 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the stromal cells are fibroblasts.
Claim 48
Dependent Claim
35 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the stromal cells are a combination of fibroblasts and endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma Cells, mast cells or adipocytes.
Claim 49
Dependent Claim
22 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the framework is composed of a biodegradable material.
Claim 50
Dependent Claim
28 words
The method for diagnosing or monitoring a malignancy according to claim 44 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 51
Dependent Claim
22 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the framework is composed of a non-biodegradable material.
Claim 52
Dependent Claim
38 words
The method for diagnosing or monitoring a malignancy according to claim 46 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 53
Dependent Claim
24 words
The method for diagnosing or monitoring a malignancy according to claim 44, 45, 46 or 47 in which the framework is pre-coated with collagen.
Claim 54
Dependent Claim
26 words
The method for diagnosing or monitoring a malignancy according to claim 41, 42, 43, 44, 45, 46 or 47 in which the framework is a mesh.
Claim 55
Dependent Claim
13 words
The method according to claim 48 in which the framework is a mesh.
Claim 56
Dependent Claim
19 words
The method for diagnosing or monitoring malignancy according to claim 41 in which the tumor cells are hematopoietic cells.
Claim 57
Dependent Claim
22 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the tumor cells are melanocyte or keratinocyte cells.
Claim 58
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the tumor cells are liver parenchymal cells.
Claim 59
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the tumor cells are pancreatic acinar cells.
Claim 60
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the tumor cells are kidney parenchymal cells.
Claim 61
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 41 in which the tumor cells are mucosal epithelium cells.
Claim 62
Independent Claim
112 words
A method for diagnosing or monitoring a malignancy in a patient comprising: (a) inoculating astrocytes from a tumor cell sample obtained from the patient onto a living stromal tissue prepared in vitro comprising confluent endothelial 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 endothelial cells; (b) inoculating further onto said astrocytes, neuron cells from the sample obtained from the patient; (c) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant astrocyte cells in the proliferated cells in culture.
Claim 63
Independent Claim
112 words
A method for diagnosing or monitoring a malignancy in a patient comprising: (a) inoculating astrocytes from a tumor cell sample obtained from the patient onto a living stromal tissue prepared in vitro comprising confluent endothelial 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 endothelial cells; (b) inoculating further onto said astrocytes, neuron cells from the sample obtained from the patient; (c) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant astrocyte cells in the proliferated cells in culture.
Claim 64
Independent Claim
111 words
A method for diagnosing or monitoring a malignancy in a patient comprising (a) inoculating astrocytes from a tumor cell sample obtained from the patient onto a living stromal tissue prepared in vitro comprising confluent endothelial 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 endothelial cells; (b) inoculating further onto said astrocytes, neuron cells from the sample obtained from the patient; (c) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant neuron cells in the proliferated cells in culture.
Claim 65
Independent Claim
111 words
A method for diagnosing or monitoring a malignancy in a patient comprising (a) inoculating astrocytes from a tumor cell sample obtained from the patient onto a living stromal tissue prepared in vitro comprising confluent endothelial 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 endothelial cells; (b) inoculating further onto said astrocytes, neuron cells from the sample obtained from the patient; (c) culturing the inoculated living stromal tissue in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant neuron cells in the proliferated cells in culture.
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