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

A bioengineering and construction method technology, applied in the field of bioengineering, can solve the problems of biosafety uncertainty, insufficient biomechanical strength and biocompatibility, and cannot replace donor corneal corneal transplantation, etc., to achieve good biocompatibility Effects on Sexuality and Biosecurity

Inactive Publication Date: 2018-06-08
SHANDONG PROVINCIAL HOSPITAL
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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

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

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Embodiment Construction

[0015] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0016] see figure 1 , the present invention provides the following technical solutions: a method for constructing bioengineered full-thickness human cornea in vitro, comprising the following steps:

[0017] S1) Cell culture: Cell culture includes preparation of single cells, corneal stromal cell culture and corneal endothelial cell preparation, and cell culture is carried out in an incubator containing 5% CO2, and the temperature of the incubator is maintained...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/38C12N5/071
CPCA61L27/3604A61L27/3804A61L27/3886A61L2430/16A61L2430/40C12N5/0697C12N2502/085C12N2533/90
Inventor 张凯王鸿李萍刘卫卫王珏徐艳云
Owner SHANDONG PROVINCIAL HOSPITAL
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