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
The object of the present invention is to provide the following: a method for preparing a basement membrane which is extracellular matrices having a function to control morphology, differentiation, proliferation, motility, function expression and the like of cells; a method for constructing a specimen of a basement membrane; a process for producing a reconstituted artificial tissue; a basement membrane specimen or an artificial tissue which can be transplanted while maintaining the structure of a basement membrane. Abasement 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 which can localize a receptor having 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 having an ability to form a basement membrane on the basement membrane specimen constructed by the following process: 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; the lipid component of cells is lysed; 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. A protein support structure is temporarily adhered to plastic surface through an adsorptive polymer by hydrophobic bonding, such as an alternating copolymer of methyl vinyl ether and maleic anhydride, which has a hydrophobic linear carbon skeleton and a functional group which can react with protein in a molecule and a basement membrane specimen or an artificial tissue is formed thereon, then the protein support structure supporting a basement membrane specimen or an artificial tissue is physically exfoliated from plastic surface when desired.
