In-vitro construction method of bioengineering full-layer human cornea

Abstract

The invention discloses an in-vitro construction method of bioengineering full-layer human cornea. Cell culture comprises preparation of single cell, cultivation of corneal stroma cells and preparation of corneal endothelial cells. The method comprises the following steps: washing a corneosclera ring of a donor with 100 U / ml penicillin and 80 U / ml streptomycin solution wash for three times, soaking into a 2.4 U / ml Dispase 2 solution, performing incubation in an incubator for 1 hour, lightly stripping limbal epithelium through a dissecting microscope and microscopic tweezers, soaking the stripped epithelial sheet into a 0.25 percent pancreatin-0.02 percent EDTA solution, and digesting in the incubator for 5 minutes again to obtain single cell. The in-vitro construction method of the bioengineering full-layer human cornea can be applied to an in-vitro medicine toxicity experiment, more importantly, sufficient biomechanical property is achieved, traction of a suture in penetrating keratoplasty and pressure in eyeballs after operation can be tolerated, and high biocompatibility and biological security are achieved, so the bioengineering full-layer human cornea can replace the donor's cornea to be applied to corneal transplantation.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to an in vitro construction method of bioengineered full-thickness human cornea. Background technique [0002] Many researchers have used natural or synthetic polymer scaffolds and normal human keratocytes or immortalized cells to construct bioengineered full-thickness corneas that resemble natural corneas in shape and structure; Insufficiency or uncertainty of biological safety, it can only be used for drug toxicity test in vitro, but cannot replace the donor cornea for corneal transplantation. Contents of the invention [0003] The technical problem to be solved by the present invention is to overcome the existing defects and provide an in vitro construction method of bioengineered full-thickness human cornea, which can not only be used in in vitro drug toxicity tests, but more importantly, it has sufficient biomechanical properties and has Good biocompatibility and bios...

AI Technical Summary

Problems solved by technology

[0002] Many researchers have used natural or synthetic polymer scaffolds and normal human keratocytes or immortalized cells to construct bioengineered full-thickness corneas that resemble natural corneas in shape and structure; Insufficiency or uncertainty of its biological safety, it can only be used for drug toxicity test in vitro, but cannot replace the donor cornea for corneal transplantation

Images