Fibrin patch containing corneal epithelial cells and the method of manufacturing thereof
OwnerRafal BULDAK, Krzysztof PIETRYGA +3inventors
InventorsRafal BULDAK, Krzysztof PIETRYGA, Adam KONKA, Katarzyna JESSE, Joanna ZEMBALA-JOHN
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Abstract
The first object of the invention is a fibrin patch containing corneal limbal epithelial cells, characterized in that it contains a protein substrate, which is dried and rehydrated fibrin, a density of corneal limbal epithelial cells on the protein substrate is from 5,000 cells/cm2 to 70,000 cells/cm2, and the protein substrate does not contain a nutrient layer. The invention discloses also the method of the manufacturing of the patch.
Claims20
A fibrin patch containing corneal limbal epithelial cells, characterized in that it comprises a protein substrate, which is dried and rehydrated fibrin, the initial density of corneal limbal epithelial cells on the protein substrate is from 5,000 cells/cm2 to 70,000 cells/cm2, and the protein substrate does not comprise a feeder layer.
A method for manufacturing the fibrin patch, comprising following steps:
- (a) preparing a mixture for producing a protein substrate, wherein the protein is fibrinogen,
- (b) manufacturing the protein substrate,
- (c) trimming of the scaffold from the protein substrate,
- (d) applying corneal limbal epithelial cells onto the scaffold,
- (e) culturing the cells on the scaffold,
- and characterized in that,
- in step (b), the mixture for making the protein substrate is poured into the bottom mold plate and allowed to gelate at 2-25° C. for 30 min to 48 h, then the protein substrate is dried to a water content of 5-95% by weight on air at 2-25° C. for 15 min to 48 h, then the substrate is rehydrated for 30 min to 2 h and in step (d), corneal limbal epithelial cells are seeded to the protein substrate to an initial density of 5,000 cells/cm2 to 70,000 cells/cm2, and in step (e) the corneal limbal epithelial cells are cultured on the scaffold prepared in step (c) until the surface of the scaffold is entirely overgrown by corneal limbal cells.
The method according the claim 4 or 7, characterized in that the mold comprises a top and a bottom plate, where the bottom plate has a cavity for pouring the mixture for obtaining the substrate.
Dependencies are inferred from the claim text.