Ectocornea-like sheet and method of constructing the same
It is intended to provide a transplantation material applicable to ocular surface diseases with a need for ectocornea transplantation (i.e., an ectocornea-like sheet). Oral mucosal epithelial cells are inoculated onto an amnion and then cultured in the coexistence of supporter cells. When a layered structure of the oral mucosal epithelial cells is formed, the outermost layer is brought into contact with air, thereby inducing differentiation. Thus, an ectocornea-like sheet having an oral mucosal epithelial cell layer on the amnion is obtained.
1 . A corneal epithelium-like sheet, comprising a cell layer in which a layered structure of cells derived from oral mucosal epithelial cells is formed.
2 . The corneal epithelium-like sheet according to claim 1 , wherein the cells of the outermost layer are not cornified.
3 . The corneal epithelium-like sheet according to claim 1 , wherein the cells of the outermost layer are flat-shaped.
4 . The corneal epithelium-like sheet according to claim 1 , wherein the corneal epithelium-like sheet has a barrier function.
5 . A corneal epithelium transplantation sheet, comprising a cell layer in which a layered structure of cells derived from oral mucosal epithelial cells is formed on a collagen layer.
6 . The corneal epithelium transplantation sheet according to claim 5 , wherein the cells of the outermost layer of the cell layer are not cornified.
7 . The corneal epithelium transplantation sheet according to claim 5 , wherein the cells of the outermost layer of the cell layer are flat-shaped.
8 . The corneal epithelium transplantation sheet according to claim 5 , wherein the collagen layer is derived from amniotic membrane.
9 . The corneal epithelium transplantation sheet according to claim 5 , wherein the collagen layer consists of amniotic membrane from which the epithelium has been removed.
10 . The corneal epithelium transplantation sheet according to claim 5 , wherein the corneal epithelium transplantation sheet has a barrier function.
11 . A method of constructing a corneal epithelium-like sheet, the method comprising:
a) culturing oral mucosal epithelial cells on a collagen layer; and
b) when the oral mucosal epithelial cells are proliferated and a layered structure of the cells is formed, bringing the outermost layer into contact with the air.
12 . The method of constructing a corneal epithelium-like sheet according to claim 11 , wherein the step a) is carried out in coexistence of supporter cells.
13 . The method of constructing a corneal epithelium-like sheet according to claim 11 , wherein the step a) is carried out in coexistence of supporter cells and in a state in which an isolating membrane with pore size through which the supporter cells cannot pass exists between the supporter cells and the collagen layer.
14 . The method of constructing a corneal epithelium-like sheet according to claim 11 , wherein the collagen layer is derived from amniotic membrane.
15 . The method of constructing a corneal epithelium-like sheet according to claim 11 , wherein the collagen layer consists of amniotic membrane from which the epithelium has been removed.
16 . A method of constructing a corneal epithelium-like sheet, comprising the steps of:
inoculating supporter cells in a first container to form a supporter cell layer;
setting a second container, which has a bottom face made of an isolation membrane with pore size through which the supporter cells cannot pass, in the first container so that the bottom face is located in a culture medium;
forming a collagen layer on the bottom face of the second container;
inoculating oral mucosal epithelial cells on the collagen layer;
culturing the oral mucosal epithelial cells to form a layered structure of the cells; and
bringing the outermost layer of the layered structure of the oral mucosal epithelial cells into contact with the air.
17 . The method of constructing the corneal epithelium-like sheet according to claim 16 , wherein the collagen layer is derived from amniotic membrane.
18 . The method of constructing the corneal epithelium-like sheet according to claim 16 , wherein the collagen layer consists of amniotic membrane from which the epithelium has been removed.
19 . The corneal epithelium-like sheet according to claim 2 , wherein the corneal epithelium-like sheet has a barrier function.
20 . The corneal epithelium-like sheet according to claim 3 , wherein the corneal epithelium-like sheet has a barrier function.