EP0358506A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Three-dimensional cell and tissue culture system
Assignee: TJ Smith and Nephew Ltd
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Filing: 1989-09-07
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Granted: —
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Expires: 2009-09-07
Abstract
Abstract The present invention relates to a three-dimensional cell culture system which 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
EP
Data Source
google_patents
Last Synchronized
2026-10-23 19:15
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: 166
Claim 1
Independent Claim
48 words
A three-dimensional stromal matrix comprising subconfluent stromal cells on a three dimensional matrix composed of a material and shape that allows cells to attach to it or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 2
Dependent Claim
48 words
A three-dimensional stromal matrix comprising subconfluent stromal cells on a three dimensional matrix composed of a material and shape that allows cells to attach to it or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 3
Dependent Claim
14 words
The three-dimensional stromal matrix of claim 1 in which the stromal cells comprise fibroblasts.
Claim 4
Dependent Claim
26 words
The three-dimensional stromal matrix of claim 2 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 5
Dependent Claim
16 words
The three-dimensional stromal matrix of claim 1 in which the three-dimensional matrix comprises a biodegradable material.
Claim 6
Dependent Claim
23 words
The three-dimensional stromal matrix of claim 4 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 7
Dependent Claim
17 words
The three-dimensional stromal matrix of claim 1 in which the three dimensional matrix comprises a non-biodegradable material.
Claim 8
Dependent Claim
33 words
The three-dimensional stromal matrix of claim 6 in which the non-biodegradable material comprises 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 stromal matrix of claim 4, 5, 6 or 7 in which the material is coated with collagen.
Claim 10
Dependent Claim
56 words
A three-dimensional cell culture comprising parenchymal cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 11
Dependent Claim
56 words
A three-dimensional cell culture comprising parenchymal cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 12
Dependent Claim
14 words
The three-dimensional cell culture of claim 9 in which the stromal cells comprise fibroblasts.
Claim 13
Dependent Claim
26 words
The three-dimensional cell culture of claim 10 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 14
Dependent Claim
16 words
The three-dimensional cell culture of claim 9 in which the three-dimensional matrix comprises a biodegradable material.
Claim 15
Dependent Claim
23 words
The three-dimensional cell culture of claim 12 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 16
Dependent Claim
17 words
The three-dimensional cell culture of claim 9 in which the three dimensional matrix comprises a non-biodegradable material.
Claim 17
Dependent Claim
33 words
The three-dimensional cell culture of claim 14 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 18
Dependent Claim
19 words
The three-dimensional cell culture of claim 12, 13, 14 or 15 in which the material is coated with collagen.
Claim 19
Dependent Claim
57 words
A three-dimensional bone marrow culture comprising hematopoietic cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 20
Dependent Claim
57 words
A three-dimensional bone marrow culture comprising hematopoietic cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 21
Dependent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 22
Dependent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 23
Dependent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 24
Dependent Claim
57 words
A three-dimensional skin culture comprising melanocytes and keratinocytes cultured on a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 25
Dependent Claim
61 words
A three-dimensional culture system comprising neuronal cells and astrocytes cultured on a three-dimensional stromal matrix comprising confluent small blood vessel endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 26
Dependent Claim
61 words
A three-dimensional culture system comprising neuronal cells and astrocytes cultured on a three-dimensional stromal matrix comprising confluent small blood vessel endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 27
Dependent Claim
56 words
A three-dimensional culture comprising mucosal epithelial cells cultured on a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 28
Dependent Claim
56 words
A three-dimensional culture comprising mucosal epithelial cells cultured on a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 29
Dependent Claim
56 words
A three-dimensional tumor culture comprising tumor cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 30
Dependent Claim
56 words
A three-dimensional tumor culture comprising tumor cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 31
Dependent Claim
55 words
A three-dimensional liver culture comprising hepatocytes cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 32
Dependent Claim
55 words
A three-dimensional liver culture comprising hepatocytes cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 33
Dependent Claim
57 words
A three-dimensional pancreatic culture comprising endocrine acinar cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 34
Dependent Claim
57 words
A three-dimensional pancreatic culture comprising endocrine acinar cells cultured on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 35
Independent Claim
77 words
A method for culturing cells in vitro comprising: (a) inoculating parenchymal cells onto a three-dimensional stromal matrix comprising stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 36
Independent Claim
77 words
A method for culturing cells in vitro comprising: (a) inoculating parenchymal cells onto a three-dimensional stromal matrix comprising stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 37
Dependent Claim
13 words
The method according to claim 25 in which the stromal cells comprise fibroblasts.
Claim 38
Dependent Claim
25 words
The method according to claim 26 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 39
Dependent Claim
15 words
The method according to claim 25 in which the three-dimensional matrix comprises a biodegradable material.
Claim 40
Dependent Claim
22 words
The method according to claim 28 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 41
Dependent Claim
16 words
The method according to claim 25 in which the three dimensional matrix comprises a non-biodegradable material.
Claim 42
Dependent Claim
32 words
The method according to claim 30 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 43
Dependent Claim
18 words
The method according to claim 28, 29, 30 or 31 in which the material is coated with collagen.
Claim 44
Independent Claim
71 words
A method for culturing bone marrow cells in vitro comprising: (a) inoculating hematopoietic cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 45
Independent Claim
71 words
A method for culturing bone marrow cells in vitro comprising: (a) inoculating hematopoietic cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 46
Independent Claim
72 words
A method for culturing skin cells in vitro , comprising: (a) inoculating melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 47
Independent Claim
72 words
A method for culturing skin cells in vitro , comprising: (a) inoculating melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 48
Independent Claim
72 words
A method for culturing skin cells in vitro , comprising: (a) inoculating melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 49
Independent Claim
72 words
A method for culturing skin cells in vitro , comprising: (a) inoculating melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 50
Independent Claim
69 words
A method for culturing neuronal cells in vitro (a) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and further (b) inoculating onto said astrocytes, neuronal cells; and (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium.
Claim 51
Independent Claim
69 words
A method for culturing neuronal cells in vitro (a) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and further (b) inoculating onto said astrocytes, neuronal cells; and (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium.
Claim 52
Independent Claim
71 words
A method for culturing mucosal epithelium in vitro comprising: (a) inoculating mucosal epithelial cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 53
Independent Claim
71 words
A method for culturing mucosal epithelium in vitro comprising: (a) inoculating mucosal epithelial cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 54
Independent Claim
71 words
A method for culturing tumor tissue cells in vitro comprising: (a) inoculating tumor cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 55
Independent Claim
71 words
A method for culturing tumor tissue cells in vitro comprising: (a) inoculating tumor cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 56
Independent Claim
69 words
A method for culturing liver cells in vitro comprising: (a) inoculating hepatocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 57
Independent Claim
69 words
A method for culturing liver cells in vitro comprising: (a) inoculating hepatocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 58
Independent Claim
71 words
A method for culturing pancreatic cells in vitro comprising: (a) inoculating pancreatic acinar cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 59
Independent Claim
71 words
A method for culturing pancreatic cells in vitro comprising: (a) inoculating pancreatic acinar cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 60
Independent Claim
44 words
A method for transplantation or implantation of cells in vivo comprising: (a) culturing parenchymal cells derived from a tissue type to be transplanted or implanted so that the parenchymal cells proliferate in vitro ; and (b) implanting the proliferated parenchymal cells in vivo .
Claim 61
Independent Claim
44 words
A method for transplantation or implantation of cells in vivo comprising: (a) culturing parenchymal cells derived from a tissue type to be transplanted or implanted so that the parenchymal cells proliferate in vitro ; and (b) implanting the proliferated parenchymal cells in vivo .
Claim 62
Dependent Claim
85 words
The method for transplantation or implantation according to claim 41 in which the culturing comprises: (a) inoculating parenchymal cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inocuated cells proliferate in culture.
Claim 63
Dependent Claim
13 words
The method according to claim 42 in which the stromal cells comprise fibroblasts.
Claim 64
Dependent Claim
25 words
The method according to claim 43 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 65
Dependent Claim
15 words
The method according to claim 42 in which the three-dimensional matrix comprises a biodegradable material.
Claim 66
Dependent Claim
22 words
The method according to claim 45 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.
Claim 67
Dependent Claim
16 words
The method according to claim 42 in which the three dimensional matrix comprises a non-biodegradable material.
Claim 68
Dependent Claim
32 words
The method according to claim 47 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 69
Dependent Claim
18 words
The method according to claim 45, 46, 47 or 48 in which the material is coated with collagen.
Claim 70
Independent Claim
35 words
A method for transplantation or implantation of bone marrow cells in vivo comprising: (a) culturing hematopoietic cells in vitro so that the hematopoietic cells proliferate; and (b) implanting the proliferated hematopoietic cells in vivo .
Claim 71
Independent Claim
35 words
A method for transplantation or implantation of bone marrow cells in vivo comprising: (a) culturing hematopoietic cells in vitro so that the hematopoietic cells proliferate; and (b) implanting the proliferated hematopoietic cells in vivo .
Claim 72
Dependent Claim
91 words
The method for transplantation or implantation of bone marrow cells according to claim 50 in which the culturing comprises: (a) inoculating hematopoietic cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 73
Independent Claim
37 words
A method for transplantation or implantation of skin cells in vivo comprising: (a) culturing melanocytes and keratinocytes in vitro so that the melanocytes and keratinocytes proliferate; and (b) implanting the proliferated melanocytes and keratinocytes in vivo .
Claim 74
Independent Claim
37 words
A method for transplantation or implantation of skin cells in vivo comprising: (a) culturing melanocytes and keratinocytes in vitro so that the melanocytes and keratinocytes proliferate; and (b) implanting the proliferated melanocytes and keratinocytes in vivo .
Claim 75
Dependent Claim
90 words
The method for transplantation or implantation of skin cells according to claim 52 in which the culturing comprises: (a) inoculating the melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 76
Dependent Claim
90 words
The method for transplantation or implantation of skin cells according to claim 52 in which the culturing comprises: (a) inoculating the melanocytes and keratinocytes onto a three-dimensional stromal matrix comprising confluent stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 77
Independent Claim
31 words
A method for transplantation or implantation of liver cells in vivo comprising: (a) culturing hepatocytes in vitro so that the hepatocytes proliferate; and (b) implanting the proliferated hepatocytes in vivo .
Claim 78
Independent Claim
31 words
A method for transplantation or implantation of liver cells in vivo comprising: (a) culturing hepatocytes in vitro so that the hepatocytes proliferate; and (b) implanting the proliferated hepatocytes in vivo .
Claim 79
Dependent Claim
88 words
The method for transplantation or implantation of liver cells according to claim 55 in which the culturing comprises: (a) inoculating the hepatocytes onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 80
Independent Claim
37 words
A method for transplantation or implantation of pancreatic cells in vivo comprising: (a) culturing pancreatic acinar cells in vitro so that the pancreatic acinar cells proliferate; and (b) implanting the proliferated pancreatic acinar cells in vivo .
Claim 81
Independent Claim
37 words
A method for transplantation or implantation of pancreatic cells in vivo comprising: (a) culturing pancreatic acinar cells in vitro so that the pancreatic acinar cells proliferate; and (b) implanting the proliferated pancreatic acinar cells in vivo .
Claim 82
Dependent Claim
90 words
The method for transplantation or implantation of pancreatic cells according to claim 57 in which the culturing comprises: (a) inoculating the pancreatic acinar cells onto a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture.
Claim 83
Independent Claim
57 words
A method for implantation of stromal cells in vivo comprising implanting a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 84
Independent Claim
57 words
A method for implantation of stromal cells in vivo comprising implanting a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer.
Claim 85
Dependent Claim
15 words
The method of implantation according to claim 59 in which the stromal cells comprise fibroblasts.
Claim 86
Dependent Claim
27 words
The method of implantation according to claim 60 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 87
Dependent Claim
17 words
The method of implantation according to claim 59 in which the three-dimensional matrix comprises a biodegradable material.
Claim 88
Dependent Claim
24 words
The method of implantation according to claim 62 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 89
Dependent Claim
16 words
The method of implantation according to claim 59 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 90
Dependent Claim
34 words
The method of implantation according to claim 64 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 91
Independent Claim
20 words
The method of implantation according to claims 62, 63, 64 or 65 in which the material is coated with collagen.
Claim 92
Independent Claim
111 words
A method for testing the cytotoxicity 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 three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cell to grow in more than one layer; and (b) determining the number of damaged or dead cells in the cell culture, in which the number of damaged or dead cells correlates with the cytotoxicity of the test substance.
Claim 93
Independent Claim
111 words
A method for testing the cytotoxicity 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 three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cell to grow in more than one layer; and (b) determining the number of damaged or dead cells in the cell culture, in which the number of damaged or dead cells correlates with the cytotoxicity of the test substance.
Claim 94
Dependent Claim
21 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the stromal cells comprise fibroblasts.
Claim 95
Dependent Claim
33 words
The method for testing the cytotoxicity of a test substance according to claim 68 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 96
Dependent Claim
23 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the three-dimensional matrix comprises a biodegradable material.
Claim 97
Dependent Claim
30 words
The method for testing the cytotoxicity of a test substance according to claim 70 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 98
Dependent Claim
22 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 99
Dependent Claim
40 words
The method for testing the cytotoxicity of a test substance according to claim 72 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 100
Dependent Claim
26 words
The method for testing the cytotoxicity of a test substance according to claim 70, 71, 72 or 73 in which the material is coated with collagen.
Claim 101
Dependent Claim
22 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the parenchymal cells comprise hematopoietic cells.
Claim 102
Dependent Claim
23 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the parenchymal cells comprise melanocytes and keratinocytes.
Claim 103
Dependent Claim
21 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the parenchymal cells comprise hepatocytes.
Claim 104
Dependent Claim
23 words
The method for testing the cytotoxicity of a test substance according to claim 67 in which the parenchymal cells comprise pancreatic acinar cells.
Claim 105
Independent Claim
83 words
A method for testing the effect of a drug, comprising: (a) exposing a three-dimensional cell culture to the drug, in which the three dimensional cell culture comprises parenchymal cells grown on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to grow in more than one layer; and (b) determining the effect of the drug on the cells in culture.
Claim 106
Independent Claim
83 words
A method for testing the effect of a drug, comprising: (a) exposing a three-dimensional cell culture to the drug, in which the three dimensional cell culture comprises parenchymal cells grown on a three-dimensional stromal matrix comprising subconfluent stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to grow in more than one layer; and (b) determining the effect of the drug on the cells in culture.
Claim 107
Dependent Claim
20 words
The method for testing the effect of a drug according to claim 79 in which the stromal cells comprise fibroblasts.
Claim 108
Dependent Claim
32 words
The method for testing the effect of a drug according to claim 80 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 109
Dependent Claim
22 words
The method for testing the effect of a drug according to claim 79 in which the three-dimensional matrix comprises a biodegradable material.
Claim 110
Dependent Claim
29 words
The method for testing the effect of a drug according to claim 82 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 111
Dependent Claim
21 words
The method for testing the effect of a drug according to claim 79 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 112
Dependent Claim
39 words
The method for testing the effect of a drug according to claim 84 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 113
Independent Claim
18 words
The method for testing the 82, 83, 84 or 85 in which the material is coated with collagen.
Claim 114
Independent Claim
18 words
The method for testing the 82, 83, 84 or 85 in which the material is coated with collagen.
Claim 115
Dependent Claim
21 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise hematopoietic cells.
Claim 116
Dependent Claim
22 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise melanocytes and keratinocytes.
Claim 117
Dependent Claim
20 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise hepatocytes.
Claim 118
Dependent Claim
22 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise pancreatic acinar cells.
Claim 119
Dependent Claim
22 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise mucosal epithelial cells.
Claim 120
Dependent Claim
26 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise mucosal epithelial cells infected with a virus.
Claim 121
Dependent Claim
26 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise mucosal epithelial cells infected with a herpesvirus.
Claim 122
Dependent Claim
26 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise mucosal epithelial cells infected with a papillomavirus.
Claim 123
Dependent Claim
21 words
The method for testing the effect of a drug according to claim 79 in which the parenchymal cells comprise tumor cells.
Claim 124
Independent Claim
109 words
A method for diagnosing or monitoring a malignancy in a patient, comprising: (a) obtaining a sample of cells from the patient; (b) inoculating cells from the sample onto a three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant cells in the proliferated cells in culture.
Claim 125
Independent Claim
109 words
A method for diagnosing or monitoring a malignancy in a patient, comprising: (a) obtaining a sample of cells from the patient; (b) inoculating cells from the sample onto a three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture; and (d) determining the presence of malignant cells in the proliferated cells in culture.
Claim 126
Dependent Claim
19 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the stromal cells comprise fibroblasts.
Claim 127
Dependent Claim
31 words
The method for diagnosing or monitoring a malignancy according to claim 97 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 128
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the three-dimensional matrix comprises a biodegradable material.
Claim 129
Dependent Claim
28 words
The method for diagnosing or monitoring a malignancy according to claim 99 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 130
Dependent Claim
20 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 131
Dependent Claim
38 words
The method for diagnosing or monitoring a malignancy according to claim 101 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 132
Dependent Claim
24 words
The method for diagnosing or monitoring a malignancy according to claim 99, 100, 101 or 102 in which the material is coated with collagen.
Claim 133
Dependent Claim
20 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the parenchymal cells comprise hematopoietic cells.
Claim 134
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the parenchymal cells comprise melanocytes and keratinocytes.
Claim 135
Dependent Claim
19 words
The method for diagnosing or monitoring a malignancy according to claim 96 in which the parenchymal cells comprise hepatocytes.
Claim 136
Dependent Claim
21 words
The method for diagnosing or monitoring a malignancy according to claim 82 in which the parenchymal cells comprise pancreatic acinar cells.
Claim 137
Independent Claim
107 words
A method for culturing genetically engineered cells, comprising: (a) inoculating transfected parenchymal cells onto a three-dimensional stromal matrix in which: (i) the transfected parenchymal cells contain a foreign gene under the control of an expression element; and (ii) the three-dimensional stromal matrix comprises actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three dimensional stromal matrix in a nutrient medium so that the transfected cells proliferate in culture.
Claim 138
Independent Claim
107 words
A method for culturing genetically engineered cells, comprising: (a) inoculating transfected parenchymal cells onto a three-dimensional stromal matrix in which: (i) the transfected parenchymal cells contain a foreign gene under the control of an expression element; and (ii) the three-dimensional stromal matrix comprises actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; and (b) incubating the inoculated three dimensional stromal matrix in a nutrient medium so that the transfected cells proliferate in culture.
Claim 139
Dependent Claim
18 words
The method for culturing genetically engineered cells according to claim 108 in which the stromal cells comprise fibroblasts.
Claim 140
Dependent Claim
30 words
The method for culturing genetically engineered cells according to claim 109 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 141
Dependent Claim
20 words
The method for culturing genetically engineered cells according to claim 108 in which the three-dimensional matrix comprises a biodegradable material.
Claim 142
Dependent Claim
27 words
The method for culturing genetically engineered cells according to claim 111 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 143
Dependent Claim
19 words
The method for culturing genetically engineered cells according to claim 108 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 144
Dependent Claim
37 words
The method for culturing genetically engineered cells according to claim 113 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 145
Dependent Claim
23 words
The method for culturing genetically engineered cells according to claim 111, 112, 113 or 114 in which the material is coated with collagen.
Claim 146
Dependent Claim
19 words
The method for culturing genetically engineered cells according to claim 108 in which the parenchymal cells comprise hematopoietic cells.
Claim 147
Dependent Claim
20 words
The method for culturing genetically engineered cells according to claim 108 in which the parenchymal cells comprise melanocytes and keratinocytes.
Claim 148
Dependent Claim
18 words
The method for culturing genetically engineered cells according to claim 108 in which the parenchymal cells comprise hepatocytes.
Claim 149
Dependent Claim
20 words
The method for culturing genetically engineered cells according to claim 108 in which the parenchymal cells comprise pancreatic acinar cells.
Claim 150
Independent Claim
110 words
A method for analyzing a disease or condition in a patient comprising: (a) obtaining a sample of cells from the patient; (b) inoculating cells from the sample onto a three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture; and (d) analyzing the proliferated cells in culture for markers of the disease or condition.
Claim 151
Independent Claim
110 words
A method for analyzing a disease or condition in a patient comprising: (a) obtaining a sample of cells from the patient; (b) inoculating cells from the sample onto a three-dimensional stromal matrix comprising actively growing stromal cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, or can be modified to allow cells to attach to it, and which allows cells to grow in more than one layer; (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium so that the inoculated cells proliferate in culture; and (d) analyzing the proliferated cells in culture for markers of the disease or condition.
Claim 152
Dependent Claim
17 words
The method for studying disease mechanisms according to claim 120 in which the stromal cells comprise fibroblasts.
Claim 153
Dependent Claim
29 words
The method for studying disease mechanisms according to claim 121 in which the stromal cells further comprise endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.
Claim 154
Dependent Claim
19 words
The method for studying disease mechanisms according to claim 120 in which the three-dimensional matrix comprises a biodegradable material.
Claim 155
Dependent Claim
26 words
The method for studying disease mechanisms according to claim 123 in which the biodegradable material comprises cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.
Claim 156
Dependent Claim
18 words
The method for studying disease mechanisms according to claim 120 in which the three-dimensional matrix comprises non-biodegradable materials.
Claim 157
Dependent Claim
36 words
The method for studying disease mechanisms according to claim 125 in which the non-biodegradable material comprises a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.
Claim 158
Dependent Claim
22 words
The method for studying disease mechanisms according to claim 123, 124, 125 or 126 in which the material is coated with collagen.
Claim 159
Dependent Claim
18 words
The method for studying disease mechanisms according to claim 120 in which the parenchymal cells comprise hematopoietic cells.
Claim 160
Dependent Claim
19 words
The method for studying disease mechanisms according to claim 120 in which the parenchymal cells comprise melanocytes and keratinocytes.
Claim 161
Dependent Claim
17 words
The method for studying disease mechanisms according to claim 120 in which the parenchymal cells comprise hepatocytes.
Claim 162
Dependent Claim
19 words
The method for studying disease mechanisms according to claim 120 in which the parenchymal cells comprise pancreatic acinar cells.
Claim 163
Independent Claim
114 words
A method for testing the ability of a substance to cross the blood-brain barrier comprising: (a) exposing an effective amount of the substance to the endothelial surface of a three-dimensional culture system made by (i) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; (ii) inoculating further, onto said astrocytes, neuronal cells; and (iii) incubating the inoculated three-dimensional stromal matrix in a nutrient medium; and (b) detecting the effects of the substance on the astrocytes or neuronal cells of the inoculated three-dimensional stromal matrix.
Claim 164
Independent Claim
114 words
A method for testing the ability of a substance to cross the blood-brain barrier comprising: (a) exposing an effective amount of the substance to the endothelial surface of a three-dimensional culture system made by (i) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; (ii) inoculating further, onto said astrocytes, neuronal cells; and (iii) incubating the inoculated three-dimensional stromal matrix in a nutrient medium; and (b) detecting the effects of the substance on the astrocytes or neuronal cells of the inoculated three-dimensional stromal matrix.
Claim 165
Dependent Claim
78 words
A model system for the blood-brain barrier comprising a three-dimensional tissue culture prepared by the following method: (a) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and further (b) inoculating onto said astrocytes, neuronal cells; and (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium.
Claim 166
Dependent Claim
78 words
A model system for the blood-brain barrier comprising a three-dimensional tissue culture prepared by the following method: (a) inoculating astrocytes onto a three-dimensional stromal matrix comprising confluent endothelial cells on a three-dimensional matrix composed of a material and shape that allows cells to attach to it, and which allows cells to grow in more than one layer; and further (b) inoculating onto said astrocytes, neuronal cells; and (c) incubating the inoculated three-dimensional stromal matrix in a nutrient medium.
References Cited (Backward Citations)
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