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AssigneeAdvanced Tissue Sciences, Inc.
InventorsGail K. Naughton, Brian A. Naughton
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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.
A genetically engineered living stromal tissue prepared in vitro, comprising stromal cells transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed, in which both the transfected stromal cells and connective tissue proteins naturally secreted by the transfected stromal cells are attached to and substantially envelope a framework composed of a biocompatible, nonliving material thereby forming a three-dimensional structure having interstitial spaces bridged by the transfected stromal cells.
A three-dimensional cell culture of genetically engineered dermis prepared in vitro, comprising stromal cells transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed, in which both the transfected stromal cells and connective tissue proteins naturally secreted by the transfected stromal cells are attached to and substantially envelope a framework composed of a biocompatible, non-living material thereby forming a three-dimensional structure having interstitial spaces bridged by the transfected stromal cells.
A three-dimensional cell culture comprising parenchymal cells cultured on a genetically engineered living stromal tissue prepared in vitro, comprising stromal cells transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed, in which both the transfected stromal cells and connective tissue proteins naturally secreted by the transfected stromal cells are attached to and substantially envelope a framework composed of a biocompatible, non-living material thereby forming a three-dimensional structure having interstitial spaces bridged by the transfected stromal cells.
A three-dimensional genetically engineered cell culture comprising transfected parenchymal sells cultured on a living stromal tissue prepared in vitro, in which both:
The three-dimensional genetically engineered cell culture of claim 24, 28, 29 or 30 in which the framework is pre-coated with collagen.
The three-dimensional genetically engineered cell culture of claim 24, 25, 26, 27, 28, 29, or 30 in which the framework is a mesh.
A three-dimensional genetically engineered culture system comprising neuron cells and transfected astrocytes cultured on a living stromal tissue prepared in vitro, in which both:
A three-dimensional genetically engineered culture system comprising transfected neuron cells and astrocytes cultured on a living stromal tissue prepared in vitro, in which both:
A method for implantation in vivo of a genetically engineered living stromal tissue prepared in vitro, comprising implanting the genetically engineered living stromal tissue comprising stromal cells transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed, in which both the transfected stromal cells and connective tissue proteins naturally secreted by the transfected stromal cells are attached to and substantially envelope a framework composed of a biocompatible, non-living material thereby forming a three-dimensional structure having interstitial spaces bridged by the transfected stromal cells, so that a tissue equivalent is formed in vivo.
A method of implanting in vivo a genetically engineered three-dimensional cell culture prepared in vitro, comprising implanting a culture of parenchymal cells inoculated onto a genetically engineered living stromal tissue, comprising stromal cells transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed, in which both the transfected stromal cells and connective tissue proteins naturally secreted by the transfected stromal cells are attached to and substantially envelope a framework composed of a biocompatible, non-living material thereby forming a three-dimensional structure having interstitial spaces bridged by the transfected stromal cells, so that a tissue equivalent is formed in vivo.
A method for transplantation or implantation of bone marrow cells in vivo, comprising implanting in vivo the hematopoietic cells cultured on a genetically engineered living stromal tissue prepared in vitro, in which both:
A method for transplantation or implantation of genetically engineered cells in vivo, comprising implanting in vivo transfected parenchymal cells cultured on a living stromal tissue prepared in vitro in which both:
The method for transplantation or implantation of genetically engineered bone marrow cells in vivo, comprising implanting in vivo the transfected hematopoietic cells cultured on a living stromal tissue prepared in vitro, in which both:
A method for transplantation or implantation of genetically engineered skin cells in vivo comprising implanting in vivo the transfected melanocyte or keratinocyte cells cultured on a living stromal tissue prepared in vitro in which both:
A method for transplantation or implantation of genetically engineered liver cells in vivo comprising implanting in vivo the transfected liver parenchymal cells cultured on a living stromal tissue prepared in vitro in which both:
A method for transplantation or implantation of genetically engineered pancreatic cells in vivo comprising implanting in vivo the transfected pancreatic acinar cells cultured on a living stromal tissue prepared in vitro in which both:
A method for transplantation or implantation of genetically engineered kidney cells in vivo comprising transplanting in vivo the transfected kidney parenchymal cells cultured on a living stromal tissue prepared in vitro in which both:
A method for transplantation or implantation of genetically engineered mucosal epithelium cells in vivo comprising implanting in vivo the transfected mucosal epithelium cells cultured on a living stromal tissue prepared in vitro in which both:
A method for transplantation or implantation of a genetically engineered blood-brain barrier cell culture in vivo comprising transplanting in vivo the neuron cells and transfected astrocytes cultured on a living stromal tissue prepared in vitro, in which both:
A method for transplantation or implantation of a three-dimensional genetically engineered blood-brain barrier cell culture in vivo comprising implanting in vivo the transfected neuron cells and astrocytes cultured on a living stromal tissue prepared in vitro, in which both:
Dependencies are inferred from the claim text.