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.