Preservation method for corneal limbus tissue

A preservation method and limbal technology, applied in the preservation, application, and animal husbandry of human or animal bodies, can solve the problems of losing epithelium, restricting the clinical application of limbal active tissue, and damaging the basement membrane, so as to achieve complete preservation and high The effect of activity and simple production process

Active Publication Date: 2014-11-19
XIAMEN EYE CENTER OF XIAMEN UNIVERSITY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During storage, traditional corneal materials will lose epithelium and even damage the basement membrane, which limits the clinical application of limbal active tissue to a certain extent.

Method used

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  • Preservation method for corneal limbus tissue
  • Preservation method for corneal limbus tissue
  • Preservation method for corneal limbus tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The preparation method of the present invention includes the following steps: the remaining limbal tissue of the corpse-derived eyeball after corneal transplantation is cleaned with sterile normal saline in a sterile and clean environment, and the blood stains on the surface are cleaned and disinfected. Fresh corneal limbus tissues are soaked and rinsed with antibiotic-containing phosphate buffer for 5-10min, rinse at least 3 times, each time greater than or equal to 5min; the surgeon wears sterile gloves with both hands and uses microscopic instruments to remove the conjunctival iris For tissue, place the sterile transwell chamber in a sterile cell culture dish with a diameter of 35mm, and spread the limbal tissue on the PVDF membrane of the transwell chamber; add the mid-term preservation solution to the bottom of the transwell chamber until the liquid surface is fully covered with the PVDF membrane Contact: Place the cell culture dish in the refrigerator to keep the am...

Embodiment 2

[0047] The preparation method of the present invention includes the following steps: the remaining limbal tissue of the corpse-derived eyeball after corneal transplantation is cleaned with sterile normal saline in a sterile and clean environment, and the blood stains on the surface are cleaned and disinfected. Fresh corneal limbus tissues are soaked and rinsed with antibiotic-containing phosphate buffer for 5-10min, rinse at least 3 times, each time greater than or equal to 5min; the surgeon wears sterile gloves with both hands and uses microscopic instruments to remove the conjunctival iris For tissue, place the sterile transwell chamber in a sterile cell culture dish with a diameter of 35mm, and spread the limbal tissue on the PVDF membrane of the transwell chamber; add the mid-term preservation solution to the bottom of the transwell chamber until the liquid surface is fully covered with the PVDF membrane Contact: Place the cell culture dish in a thermostat, keep the ambient ...

Embodiment 3

[0049] The preparation method of the present invention includes the following steps: the remaining limbal tissue of the corpse-derived eyeball after corneal transplantation is cleaned with sterile normal saline in a sterile clean environment, and the blood stains on the surface are cleaned and disinfected. Fresh corneal limbus tissues are soaked and rinsed with antibiotic-containing phosphate buffer for 5-10min, rinse at least 3 times, each time greater than or equal to 5min; the surgeon wears sterile gloves with both hands and uses microscopic instruments to remove the conjunctival iris For tissue, place the sterile transwell chamber in a sterile cell culture dish with a diameter of 35mm, and spread the limbal tissue on the PVDF membrane of the transwell chamber; add SHEM medium to the bottom of the transwell chamber until the liquid surface is fully connected to the PVDF membrane Contact: Put the cell culture dish in the refrigerator to keep the ambient temperature constant at...

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Abstract

The invention discloses a preservation method for corneal limbus tissue. The method comprises a step of disinfecting fresh corneal limbus tissue and a step of air exposure preservation of the disinfected fresh corneal limbus tissue. The step of air exposure preservation is as follows: the fresh corneal limbus tissue is placed on a permeable covering film, preservation liquid or a cell culture medium capable of providing nutrients to the corneal limbus tissue is arranged below the covering film, epitheliums of cornea directly contact with air, the temperature of a culture environment in the step is 0 to 37 DEG C, and oxygen concentration is 0 to 20%. Compared to immersion preservation in the prior art, the preservation method provided in the invention has the following advantages: the structure of the preserved corneal limbus tissue is more integral; obvious apoptosis of the epitheliums of cornea does not occur; stem cells of corneal limbus can maintain high activity; and process flow is simple and reliable, is easy to implement, costs little and has other efficacy.

Description

Technical field [0001] The invention relates to the field of preservation of graft materials, and more specifically to a method for preservation of limbal tissue. Background technique [0002] Limbal tissue is widely used in many diseases such as limbal stem cell deficiency or limbal stem cell dysfunction, limbal corneal degeneration and limbal tumors. One of the important issues in this treatment is to remove the limbal tissue from the donor and keep it alive. [0003] In 1992, Kaufman reported the Optisol corneal active preservation solution, which makes the preservation of corneal slices in eye banks safe and reliable within 2-3 weeks, and has become the first choice for penetrating corneal transplantation in eye banks around the world. In China, 2-4° wet room preservation method is mainly used. Traditional corneal transplantation takes penetrating keratoplasty (PKP) as the mainstream surgical procedure. In order to consider corneal grafts to meet penetrating keratoplasty, it ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 吴护平董诺李程刘祖国
Owner XIAMEN EYE CENTER OF XIAMEN UNIVERSITY CO LTD
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