Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
90 words
A composite living skin equivalent which comprises relative to a horizontal plane: a) a sponge first layer comprising a cross-linked collagen sponge, said first layer having upper and lower surface, said sponge containing cultured fibroblast cells therein, b) a second layer comprising a high purity non-porous collagen, said second layer having upper and lower surfaces, the lower surface thereof being in contact with the upper surface of said first layer, and c) a layer comprising cultured keratinocyte cells in contact with the upper surface of said non-porous collagen second layer.
Claim 2
Dependent Claim
90 words
A composite living skin equivalent which comprises relative to a horizontal plane: a) a sponge first layer comprising a cross-linked collagen sponge, said first layer having upper and lower surface, said sponge containing cultured fibroblast cells therein, b) a second layer comprising a high purity non-porous collagen, said second layer having upper and lower surfaces, the lower surface thereof being in contact with the upper surface of said first layer, and c) a layer comprising cultured keratinocyte cells in contact with the upper surface of said non-porous collagen second layer.
Claim 3
Dependent Claim
18 words
The composite living skin equivalent of claim 1 wherein the collagen of the non-porous second layer is polymerized.
Claim 4
Dependent Claim
22 words
The composite living skin equivalent of claim 2 wherein the collagen of at least one of the layers is bovine source derived.
Claim 5
Dependent Claim
28 words
The composite living skin equivalent of claim 3, wherein the non-porous collagen is selected from the group consisting of Type 1 collagen, Type 3 collagen, and mixtures thereof.
Claim 6
Dependent Claim
20 words
The composite living skin equivalent of claim 4, wherein the non-porous collagen has been highly purified by pre-treatment with pepsin.
Claim 7
Dependent Claim
30 words
The composite living skin equivalent of claim 5 wherein the pores of the cross-linked first layer of collagen are of sufficient size to permit the growth of fibroblast cells therein.
Claim 8
Dependent Claim
33 words
The composite living skin equivalent of claim 6 wherein the non-porous collagen second layer is applied to said upper surface of said second layer as a thin film from a solution of collagen.
Claim 9
Dependent Claim
29 words
The composite living skin equivalent of claim 7 wherein the second layer of collagen contacts the first layer of collagen only on the upper surface of said first layer.
Claim 10
Dependent Claim
18 words
The composite living skin equivalent of claim 8 wherein the fibroblast and keratinocytes cells are human source derived.
Claim 11
Dependent Claim
21 words
The composite living skin equivalent of claim 9, wherein the fibroblast cells are obtained from treating a dermal sample with collagenase.
Claim 12
Dependent Claim
42 words
The composite living skin equivalent of claim 10, wherein the layer of keratinocyte cells is prepared by obtaining a single-cell suspension of keratinocytes, and evenly and discontinuously distributing drops of the suspension onto the layer of non-porous collagen, which is then incubated.