Method for growing cellular tissue

Claim 3

The method of claim 1 wherein the porous substrate is a microporous polymeric membrane.

Claim 4

The method of claim 2 wherein the microporous polymeric membrane is coated with collagen.

Claim 5

The method of claim 1 further comprising the steps of: a) treating a porous substrate suitable for growing cells to thereby provide activated sites for attaching growth factor thereto; b) contacting the porous substrate with cell culture medium comprising growth factor specific for growth of said cells under conditions whereby the growth factor is dispersed within the substrate; c) removing the medium from the substrate to provide a medium-free substrate; d) contacting the medium-free substrate with an amount of cell culture medium comprising growth factor and non-specific protein, sufficient to saturate remaining activated sites; e) subsequently seeding the cells onto the porous substrate; and f) maintaining the seeded porous substrate under conditions suitable for cell growth to thereby produce a confluent monolayer of tissue or uniformly differentiated multilayer tissue.

Claim 7

Tissue produced by the method of claims 1 or 5.

Claim 8

The method of claim 4 comprising the steps of: a) contacting the collagen-coated microporous substrate with cell culture medium comprising epidermal growth factor and pituitary extract containing growth factor therein, under conditions whereby the growth factors are dispersed within the substrate; b) contacting the medium-free microporous substrate with a sufficient amount of cell culture medium comprising serum, epidermal growth factor and pituitary extract containing growth factor therein, to saturate remaining activated sites; c) subsequently seeding keratinocyte cells onto the microprous substrate.

Claim 11

The method of claim 8 wherein microporous polymeric membrane substrate is a microporous membrane coated with collagen.

Claim 12

A substrate for growing tissue from cells produced by the method of claim 8.

Claim 13

A method for determining toxic effects of a substance on tissue, comprising the steps of: a) providing a tissue from keratinocyte cells which is grown on a microporous substrate suitable for growing cells having growth factors specific for growing the cells thereon produced by the method of claim 7; b) administering an amount of the substance to the tissue and observing cellular response to the substance; and c) evaluating the response to determine toxic effects of the substance.