Three-dimensional tumor cell and tissue culture system
First Claim
1. 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.
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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.
79 Citations
58 Claims
- 1. 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.
- 11. 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.
- 21. 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.
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31. A method for testing the effect of a drug comprising:
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(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. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 40)
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41. A method for diagnosing or monitoring a malignancy in a patient, comprising:
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(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. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56)
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57. A method for diagnosing or monitoring a malignancy in a patient comprising:
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(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.
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58. 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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Specification