Patent ID: 7824913

Claim:
A method for producing a cultured tissue construct, comprising, (a) seeding human fibroblast cells capable of synthesizing an extracellular matrix on a porous membrane, wherein the membrane comprises pores that are about 3 microns or less in size, in a culture vessel in a chemically defined cell culture medium, wherein the chemically defined cell culture medium is free of undefined animal organ or tissue extracts and comprises the following: (i) nutrient base medium; (ii) insulin; (iii) L-glutamine or an L-glutamine derivative; and (iv) ascorbate or an ascorbate derivative; (b) stimulating the fibroblast cells, which are at least at 80% confluence to synthesize, secrete and organize extracellular matrix components; and, (c) continued culturing of the fibroblast cells to thereby produce a cultured tissue construct with an extracellular matrix of at least about 30 microns thick and cultured fibroblast cells contained within the synthesized extracellular matrix layer, wherein the extracellular matrix comprises: (i) fibrillar collagen showing a packing organization of fibrils and fibril bundles exhibiting a quarter-staggered 67 nm banding pattern; (ii) tenascin; and, (iii) glycosaminoglycans; and wherein said extracellular matrix is produced by the cultured fibroblast cells on one surface of the porous membrane.