Method of producing retinal pigment epithelial cell sheet

Abstract

The present invention provides a method of producing a cell sheet including the following steps (1) seeding and culturing retinal pigment epithelial cells on a collagen gel to form a cell sheet composed of the retinal pigment epithelial cells, and (2) degrading the collagen gel with collagenase to detach the cell sheet composed of the retinal pigment epithelial cells, and the like.

1 . A method of producing a cell sheet comprising the following steps
 (1) seeding and culturing retinal pigment epithelial cells on a collagen gel to form a cell sheet composed of the retinal pigment epithelial cells, and   (2) degrading the collagen gel with collagenase to detach the cell sheet composed of the retinal pigment epithelial cells.   
     
     
         2 . The production method of the cell sheet according to  claim 1 , further comprising the following step (3):
 (3) confirming the presence or absence of a basement membrane on the contact surface between the detached cell sheet and the collagen gel.   
     
     
         3 . The production method of the cell sheet according to  claim 1 , wherein the retinal pigment epithelial cells are cells obtained by inducing differentiation of stem cells or progenitor cells. 
     
     
         4 . The production method of the cell sheet according to  claim 3 , further comprising the following step (3):
 (3) confirming the presence or absence of a basement membrane on the contact surface between the detached cell sheet and the collagen gel.   
     
     
         5 . The production method of the cell sheet according to  claim 3 , wherein the stem cells are ES cells or iPS cells. 
     
     
         6 . The production method of the cell sheet according to  claim 5 , further comprising the following step (3):
 (3) confirming the presence or absence of a basement membrane on the contact surface between the detached cell sheet and the collagen gel.   
     
     
         7 . The production method of the cell sheet according to  claim 1 , wherein the concentration of collagen in the collagen gel is 0.1%-0.5%. 
     
     
         8 . The production method of the cell sheet according to  claim 7 , further comprising the following step (3):
 (3) confirming the presence or absence of a basement membrane on the contact surface between the detached cell sheet and the collagen gel.   
     
     
         9 . A cell sheet produced by the method according to  claim 1 . 
     
     
         10 . A cell sheet produced by the method according to  claim 2 . 
     
     
         11 . A cell sheet produced by the method according to  claim 3 . 
     
     
         12 . A cell sheet produced by the method according to  claim 4 . 
     
     
         13 . A cell sheet produced by the method according to  claim 5 . 
     
     
         14 . A cell sheet produced by the method according to  claim 6 . 
     
     
         15 . A cell sheet produced by the method according to  claim 7 . 
     
     
         16 . A cell sheet produced by the method according to  claim 8 . 
     
     
         17 . A cell sheet for transplantation, comprising a cell layer formed with retinal pigment epithelial cells obtained by inducing differentiation of stem cells or progenitor cells ex vivo, and a basement membrane secreted from the cells. 
     
     
         18 . A cell sheet for screening, comprising a cell layer formed with retinal pigment epithelial cells obtained by inducing differentiation of stem cells or progenitor cells ex vivo, and a basement membrane secreted from said cells.