Compositions and methods for production and use of an injectable naturally secreted extracellular matrix

Claim 3

The method according to claim 1 in which the naturally secreted extracellular matrix is produced by a method comprising: (a) providing a living stromal tissue prepared in vitro comprising human stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework, said framework composed of a biocompatible, non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells; (b) killing the cells in the living stromal tissue; (c) removing the killed cells and any cellular contents from the framework; (d) collecting the connective tissue proteins naturally secreted by the stromal cells attached to the framework; and (e) processing the collected connective tissue proteins of step (d) with a pharmaceutically acceptable carrier into a formulation that is suitable for in vivo administration by injection via a syringe.

Claim 4

The method according to claim 2 in which the collected connective tissue proteins of step (d) are processed by homogenizing, cross-linking, or suspending the collected connective tissue proteins in a physiological acceptable carrier prior to step (e).

Claim 5

The method according to claim 2 in which the collected connective tissue proteins of step (d) are processed by adjusting ratios of collagen types I-V, respective to each other, prior to step (e).

Claim 6

The method according to claim 2 in which the stromal cells of the living stromal tissue are fibroblasts.

Claim 7

The method according to claim 2 in which the stromal cells of the living stromal tissue are cells found in loose connective tissue or bone marrow.

Claim 8

The method according to claim 6 in which the stromal cells of the living stromal tissue are endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells, chondrocytes or adipocytes.

Claim 9

The method according to claim 2 in which the framework is composed of a biodegradable material.

Claim 10

The method according to claim 8 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin, or dextran.

Claim 11

The method according to claim 2 in which the framework is composed of a non-biodegradable material.

Claim 12

The method 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 polytetrafluoroethylene, or a nitrocellulose compound.

Claim 13

The method according to claim 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 or 11 in which the framework is a mesh.

Claim 14

The method according to claim 12 in which the framework has pore spaces of about 150 μm to about 220 μm.

Claim 15

The method according to claim 2 in which the pharmaceutically acceptable carrier contains an anesthetic agent, a lubricating agent, a tissue growth factor or combinations thereof.

Claim 16

The method according to claim 1 in which the naturally secreted extracellular matrix is produced by a method comprising: (a) providing a living stromal tissue prepared in vitro comprising human stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework, said framework composed of a biocompatible, non-living biodegradable material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells; (b) killing the cells in the living stromal tissue; (c) removing the killed cells and any cellular contents from the framework; and (d) processing the connective tissue proteins naturally secreted by the stromal cells attached to the framework and the biodegradable framework with a pharmaceutically acceptable carrier into a formulation suitable for in vivo administration by injection via a syringe.

Claim 17

The method according to claim 6 in which the stromal cells of the living stromal tissue are glial cells, liver cells, hematopoietic cells, dermal cells, lens fiber cells, smooth muscle cells, osteoblast cells, odontoblast cells, or retinal cells.