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Full capsular bag culture regeneration crystalline lens model as well as construction method and application thereof

A lens and capsular bag technology is applied in the field of full capsular bag culture and regeneration lens model and its construction.

Pending Publication Date: 2022-05-13
EYE INST OF SHANDONG FIRST MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the prior art has studies on in vitro stem cell-induced lensoids, they are still in the stage of culturing normal lens morphology, structure and function, and they are mainly used as in vitro models for cataract-related research.
In addition, most of the existing in vitro lens culture models have damage to the capsule or lens epithelial cells, or it is difficult to maintain the original spherical structure of the lens

Method used

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  • Full capsular bag culture regeneration crystalline lens model as well as construction method and application thereof
  • Full capsular bag culture regeneration crystalline lens model as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Male C57BL / 6 mice (age, 5 weeks, body weight, 14-20 g) were purchased from Beijing Weitong Lihua Biological Co., Ltd. The mice were kept in a pathogen-free environment in the animal room of Shandong Ophthalmology Institute.

[0031]After the mice were killed by neck dislocation, the eyeballs were taken out and placed in a 1.5ml EP tube (PBS containing double antibodies) and placed in an ice box. After alcohol disinfection, the eyeballs were washed twice with PBS containing double antibodies again in the ultra-clean bench . Under a stereomicroscope, use ophthalmic scissors to cut the cornea along the limbus, separate the iris with the help of microscopic forceps, and break the zonule. Before removing the lens, use ophthalmic scissors to cut a small opening on the anterior capsule to mark the front capsule. When removing the lens, avoid surrounding cluttered tissue. After the lens was taken out, it was rinsed twice in PBS again to remove the surrounding tissue as much ...

Embodiment 2

[0036] The difference from Example 1 is that when preparing the low-melting point agarose gel, the amount of low-melting point agarose powder is 0.05 g, and during cultivation, from the first day to the seventh day, the culture medium is added bFGF on the basis of the basal medium 5ng / ml; on the 8th day to the 14th day, the medium was supplemented with bFGF 200ng / ml on the basis of the basal medium, wherein the basal medium was composed of DMEM / F-12, 1% B27, 1% L-glutamine Amide, 1% double antibody, 5% FBS, and the rest are the same as in Example 1. After culturing for 14 days, the regenerated lens was obtained.

Embodiment 3

[0038] The difference from Example 1 is that when preparing low-melting point agarose gel, the amount of low-melting point agarose powder is 0.15 g, and during cultivation, from day 1 to day 7, the culture medium is added bFGF on the basis of basal medium 3ng / ml; on the 8th day to the 14th day, the medium was supplemented with bFGF 150ng / ml on the basis of the basal medium, wherein the basal medium consisted of DMEM / F-12, 1% B27, 1% L-glutamine Amide, 1% double antibody, 5% FBS, and the rest are the same as in Example 1. After culturing for 14 days, the regenerated lens was obtained.

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Abstract

The invention provides a full capsular bag culture regeneration crystalline lens model and a construction method and application thereof, and belongs to the technical field of regeneration crystalline lens model construction. The invention provides a method for constructing a full-capsular culture regenerated crystalline lens model, which comprises the following steps of: upwards placing an anterior capsular membrane of a crystalline lens in low-melting-point agarose gel with the temperature of 37 DEG C, enabling the liquid level of the gel to submerge the equator part of the crystalline lens, reserving the anterior capsular membrane, and after the gel is solidified, cutting at the anterior capsular membrane, and removing crystalline lens fiber agglomerates; and then changing the liquid of the whole lens capsular bag fixed by the gel in the culture medium every other day for culture. According to the crystalline lens regeneration construction method, low-melting-point agarose gel is used, crystalline lens tissue can be rapidly fixed, damage to crystalline lens epithelial cells can be reduced, the 3D spherical structure of the capsule can be maintained, and the defects that an existing in-vitro crystalline lens culture model mostly damages the capsule or the crystalline lens epithelial cells, and the culture efficiency is high are overcome. Or the original spherical structure of the crystalline lens is difficult to maintain.

Description

technical field [0001] The invention belongs to the technical field of constructing a regenerative lens model, and in particular relates to a full capsular bag cultured regenerative lens model and its construction method and application. Background technique [0002] Cataract is the world's first blinding eye disease, and its only effective treatment is to remove the cloudy lens through surgery and implant an artificial lens to restore visual function. Although intraocular lenses have been widely used clinically, there are still many complications, especially for infants with cataract. Therefore, it is of great significance to explore new cataract treatment strategies for the clinical treatment of cataract. Currently, the lenses regenerated in mammals do not meet the standard of normal lenses, and the regeneration mechanism and optimization strategy are still unclear. Therefore, an effective lens culture system in vitro is necessary for the study of lens regeneration mecha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12Q1/02
CPCC12N5/0621G01N33/5088C12N2533/76C12N2501/115C12N2500/32C12N2503/02G01N2500/10
Inventor 谢立信周庆军黄钰森李宗义刘晓敏
Owner EYE INST OF SHANDONG FIRST MEDICAL UNIV
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