Middle ear mucosa-like cell sheet, process of producing the same and method of using the same

Abstract

The purpose of the present invention is to obtain an alternative to a substitute of the mucosa in the middle ear which is engrafted on the surface of the bone in the middle ear, hyperplasia of the granulation tissue and the bone and the development of the fibroblast cell sir the middle ear are suppressed, and to obtain a middle ear mucosa-like cell sheet retaining cilia in the surface layer, comprising culturing nasal epithelium cells on a cell culture substrate coated with a polymer whose hydration force changes within a temperature range of 0 to 80 ° C., wherein the cells are cultured within a temperature range where the hydration force of the polymer is weak, and then changing the temperature to a temperature at which the hydration force is strong to recover the cultured cell sheet.

1 . A middle ear mucosa-like cell sheet comprising nasal epithelium cells retaining cilia thereof, wherein the middle ear mucosa-like cell sheet has been detached from a culture substrate. 
     
     
         2 . The middle ear mucosa-like cell sheet according to  claim 1 , wherein the nasal epithelium cells are originated from mucosa in nasal cavity, in paranasal sinus or in mastoid antrum. 
     
     
         3 . The middle ear mucosa-like cell sheet according to  claim 1 , wherein the nasal epithelium cells are originated from superior turbinate mucosa, middle turbinate mucosa, inferior turbinate mucosa or mastoid antrum mucosa. 
     
     
         4 . (canceled) 
     
     
         5 . The middle ear mucosa-like cell sheet according to  claim 1 , which has a ventilatory function. 
     
     
         6 . The middle ear mucosa-like cell sheet according to  claim 1 , which has a granulation tissue inhibitory function. 
     
     
         7 . The middle ear mucosa-like cell sheet according to  claim 1 , which has a mucin secretion function. 
     
     
         8 . The middle ear mucosa-like cell sheet according to  claim 1 , which is transgenic. 
     
     
         9 . The middle ear mucosa-like cell sheet according to  claim 1 , which is in the form of a laminate comprising two or more middle ear mucosa-like cell sheets. 
     
     
         10 . The middle ear mucosa-like cell sheet according to  claim 1 , which is in the form of a laminate comprising two or more middle ear mucosa-like cell sheets, wherein each middle ear mucosa-like cell sheet comprises one or more kinds of cells selected from vascular endothelial cell, endothelial precursor cell, fat-derived cell and mesenchymal stem cell. 
     
     
         11 . A process of producing a middle ear mucosa-like cell sheet, comprising culturing nasal epithelium cells on a cell culture substrate coated with a polymer whose hydration force changes within a temperature range of 0 to 80° C., wherein the cells are cultured within a temperature range where the hydration force of the polymer is weak; and then changing the temperature to a temperature at which the hydration force is strong to detach the cultured middle ear mucosa-like cells while retaining the form of a sheet. 
     
     
         12 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the cell culture substrate is a cell insert which has a porous membrane for a cell culture surface. 
     
     
         13 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , comprising laminating the detached middle ear mucosa-like cell sheet on another middle ear mucosa-like cell sheet, or, optionally, repeating this operation, to laminate the middle ear mucosa-like cell sheets. 
     
     
         14 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the middle ear mucosa-like cell sheet is one detached from the cell culture substrate without being treated with a protease. 
     
     
         15 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the middle ear mucosa-like cell sheet is detached by intimately contacting the cultured cells with a carrier at the end of culturing, and detaching the cell sheet together with the carrier. 
     
     
         16 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the middle ear mucosa-like cells were collected from a living tissue. 
     
     
         17 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the number of the cells seeded in the culturing is 3×10 4  to 2×10 5  cells/cm 2 . 
     
     
         18 . The process of producing a middle ear mucosa-like cell sheet according to  claim 11 , wherein the polymer whose hydration force changes within a temperature range of 0 to 80° C. is poly(N-isopropylacrylamide). 
     
     
         19 . A method of using a middle ear mucosa-like cell sheet, comprising transplanting the middle ear mucosa-like cell sheet according to  claim 1  to a middle ear cavity or a mastoid antrum. 
     
     
         20 . The method of using a middle ear mucosa-like cell sheet according to  claim 19 , wherein the cells are transplanted to a transplantation site preliminarily coated with a fibrin glue. 
     
     
         21 . The method of using a middle ear mucosa-like cell sheet according to  claim 19 , which is for treating cholesteatoma of middle ear, adhesive otitis, postoperative mucocele of paranasal sinus, frontal sinus cyst, oropharyngeal cancer, oropharyngeal cancer or paranasal cancer or laryngeal cancer, or reconstruction of middle ear cavity.