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US 5,374,515 AGranted
In vitro cornea equivalent model
AssigneesOrganogenesis, Inc., President and Fellows of Harvard College
InventorsNancy L. Parenteau, Valerie S. Mason, Bjorn R. Olsen
Life of this patent
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Abstract
This invention is directed to an organ equivalent of the cornea part of the eye made using tissue culturing systems. The method of constructing the cornea equivalent results in a structure analogous to the eye cornea in vivo. The cornea equivalent is an in vitro model of the eye, which can be used for transplantation or implantation in vivo or for screening compounds in vitro.
Claims28
A cornea equivalent comprising:
- (a) an inner endothelial cell layer, wherein said endothelial cells are derived from mammalian corneal endothelial cells;
- (b) a middle stromal cell-collagen mixture layer, wherein said stromal cells are derived from mammalian corneal stromal fibroblast cells;
- and
- (c) an external epithelial cell layer, wherein said epithelial cells are derived from mammalian corneal epithelial cells.
1 more element
The method of producing a cornea equivalent comprising:
- (a) culturing endothelial cells to form an inner endothelial cell layer, wherein said endothelial cells are derived from mammalian corneal cells;
- (b) mixing stromal cells with collagen to achieve a stromal cell-collagen mixture, wherein said stromal cells are derived from mammalian corneal stromal fibroblast cells;
- (c) contacting said inner endothelial cell layer of step (a) with said stromal cell-collagen mixture of step (b), thereby forming a middle layer provided on said inner layer;
- (d) culturing said inner endothelial cell layer and said middle layer;
- (e) contacting epithelial cells onto said middle layer of step (d), wherein said epithelial cells are derived from mammalian corneal epithelial cells;
- (f) culturing said epithelial cells with said middle layer until said middle layer is covered with an external layer of epithelial cells;
- and,
- (g) continue culturing said inner, middle, and external layers under conditions to achieve a moist interface and forming a cornea equivalent.
5 more elements
A cornea equivalent comprising:
- (a) an inner endothelial cell layer, wherein said endothelial cells are derived from the mammalian endothelial cells;
- (b) a middle stromal cell-collagen mixture layer, wherein said stromal cells are mammalian stromal fibroblasts derived from the group comprising sclera, dermis, tendon or fascia;
- and
- (c) an external epithelial cell layer, wherein said epithelial cells are mammalian epithelial keratinocytes derived from the group comprising corneal keratinocytes and scleral keratinocytes.
1 more element
The method of producing a cornea equivalent comprising:
- (a) culturing mammalian endothelial cells to form an inner endothelial cell layer;
- (b) mixing mammalian stromal cells with collagen to achieve a stromal cell-collagen mixture, wherein said stromal cells are mammalian stromal fibroblasts derived from sclera, dermis, tendon or fascia;
- (c) contacting said inner endothelial cell layer of step (a) with said stromal cell-collagen mixture of step (b), thereby forming a middle layer provided on said inner layer;
- (d) culturing said inner endothelial cell layer and said middle layer;
- (e) contacting mammalian epithelial cells onto said middle layer of step (d), wherein said epithelial cells are mammalian epithelial keratinocytes selected from the group consisting of corneal keratinocytes and scleral keratinocytes;
- (f) culturing said epithelial cells with said middle layer until said middle layer is covered with an external layer of epithelial cells;
- and,
- (g) continue culturing said inner, middle, and external layers under conditions to achieve a moist interface and forming a cornea equivalent.
5 more elements
Dependencies are inferred from the claim text.