Biological Tissue Sheet, Method Of Forming The Same And Transplantation Method By Using The Sheet

1 . A biological tissue sheet comprising in vivo-derived cells proliferated on amniotic membrane in the absence of a xenogeneic animal cell.

2 . The biological tissue sheet according to claim 1 , wherein the in vivo-derived cells are proliferated in a state in which the amniotic membrane is placed on a collagen gel containing human fibroblasts.

3 . The biological tissue sheet according to claim 1 , wherein the in vivo-derived cells are proliferated by using a serum free medium.

4 . The biological tissue sheet according to claim 1 , wherein the in vivo-derived cells are proliferated by using a medium including only serum derived from a recipient as a serum component.

5 . The biological tissue sheet claim 1 , wherein the in vivo-derived cells are cells derived from corneal epithelium, conjunctival epithelium, skin epidermis, hair follicle epithelium, oral mucosa, respiratory tract mucosa or intestinal tract mucosa.

6 . The biological tissue sheet claim 1 , wherein the amniotic membrane is amniotic membrane from which epithelium has been removed.

7 . The biological tissue sheet claim 1 , comprising the amniotic membrane as a culture substrate in addition to proliferated cells.

8 . The biological tissue sheet according to claim 7 , which is a transplantation material to be transplanted to a defective tissue site via second amniotic membrane.

9 . The biological tissue sheet according to claim 1 , comprising cells obtained by placing in vivo-derived cells, which are proliferated on amniotic membrane placed on a collagen gel containing human fibroblasts, on second amniotic membrane together with the amniotic membrane in the absence of a xenogeneic animal cell, and further proliferating thereof.

10 . A forming method of a biological tissue sheet, the method comprising the following steps:

(a) preparing in vivo-derived cells;

(b) sowing the in vivo-derived cells on amniotic membrane; and

(c) culturing and proliferating the in vivo-derived cells in the absence of a xenogeneic animal cell.

11 . The forming method according to claim 10 , wherein the step (b) consists of the following steps:

(b-1) culturing human fibroblasts in a collagen gel; and

(b-2) placing amniotic membrane on the collagen gel, followed by plating the in vivo-derived cells on the amniotic membrane.

12 . The forming method according to claim 10 , further comprising the following step:

(d) after the in vivo-derived cells are proliferated, bringing the outermost surface layer into contact with air.

13 . The forming method according to claim 11 , further comprising the following steps:

(e) collecting the in vivo-derived cells together with the amniotic membrane; and

(f) placing the collected in vivo-derived cells and the amniotic membrane on second amniotic membrane with a side of the amniotic membrane facing downward, followed by culturing and proliferating the in vivo-derived cells.

14 . The forming method according to claim 10 , wherein the step (c) is carried out by using a serum free medium.

15 . The forming method according to claim 10 , wherein the step (c) is carried out by using a medium including only serum derived from a recipient as a serum component.

16 . The forming method according to claim 10 , wherein the in vivo-derived cells are cells derived from corneal epithelium, conjunctival epithelium, skin epidermis, hair follicle epithelium, oral mucosa, respiratory tract mucosa or intestinal tract mucosa.

17 . The forming method according to claim 10 , wherein the amniotic membrane is amniotic membrane from which epithelium has been removed.

18 . A method of preparing a skin epidermal cell, the method comprising the following steps:

(A) plating skin epidermal cells onto amniotic membrane;

(B) culturing and proliferating the skin epidermal cells; and

(C) collecting the proliferated skin epidermal cells.

19 . A transplantation method using the biological tissue sheet according to claim 1 as a transplantation material.