Patent ID: 8889417
Filing Date: 2014-11-18
Classification: A61L,C12N

Abstract:
1. A process for producing a cultured multi-layered cell sheet adhered to a polymer membrane, wherein the cultured multi-layered cell sheet is used for grafting and has intracellular desmosome structures and intact cell-to-substrate basal membrane protein and said cell sheet consists of mucous membrane cells, said process comprising (1) culturing cells in culture solution on a temperature-responsive polymer which has been coated on a substrate surface of a cell culture support, wherein the temperature-responsive polymer has a lower critical solution temperature in water of 0-80° C., to form said cultured multi-layered cell sheet, wherein the temperature-responsive polymer is poly(N-isopropylacrylamide), and wherein the culture solution is a medium for multi-layer culture; (2) adjusting the temperature of the culture solution to below the lower critical solution temperature, whereby the substrate surface is made hydrophilic and adhesion of the cell sheet to the substrate is weakened; (3) adhering the cultured multi-layered cell sheet to the polymer membrane; and (4) peeling the cultured multi-layered cell sheet adhered to the polymer membrane from the substrate surface to obtain the cultured multi-layered cell sheet adhered to the polymer membrane.