Method of preparing basement membrane, method of constructing basement membrane specimen, reconstituted artificial tissue using the basement membrane specimen and process for producing the same

摘要

A basement membrane having a barrier function is formed by culturing alveolar epithelial cells or vascular endothelial cells on a fibrous collagen matrix coated with a polymer having a sugar chain that can localize a receptor that has an activity to accumulate a basement membrane component on the basal surface of the cells having an ability to form a basement membrane. A reconstructed artificial tissue is obtained by seeding and culturing desired homogeneous or heterogeneous cells on the basement membrane specimen constructed by the following process: (i) the cells having an ability to form a basement membrane adhered onto a support structure through a basement membrane are treated with a surface active agent; (ii) the lipid component of cells is lysed; (iii) the mixture of an alkaline solution and a protease inhibitor is used to lyse the protein remained on the surface of the basement membrane of the cells.

主权项

1. A process for producing a reconstituted artificial tissue wherein the process comprises the following steps:
(a) seeding and culturing homogenous or heterogeneous cells having an ability to form a basement membrane to provide either (i) a basement membrane specimen having a function to control morphology, differentiation, proliferation, and motility of cells, or (ii) a basement membrane components-amorphous deposits specimen which is an incomplete basement membrane comprised of amorphous structure irregularly containing basement membrane components; (b) seeding and culturing cells having an ability to form a basement membrane on the basement membrane specimen or the basement membrane components-amorphous deposits specimen; and (c) reconstituting an artificial tissue on the basement membrane specimen or the basement membrane components-amorphous deposits specimen; wherein the basement membrane specimen or the basement membrane components-amorphous deposits specimen in step (a) is obtained from a basement membrane or a basement membrane components-amorphous deposits prepared by culturing the cells having an ability to form a basement membrane on a support structure with a sugar chain-coat that can localize a receptor; wherein the receptor has an activity to accumulate a basement membrane component onto (i) the basal surface of the cells having an ability to form a basement membrane, or (ii) the surface of the basement membrane components-amorphous deposits; and wherein the sugar-chain coat has a B-D-glucopyranosyl non-reducing end or a 2-acetoamide-2-deoxy-B-D-glucopyranosyl non-reducing end.