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Cornea mid-term preserving fluid and preparation method thereof

A preservation solution and cornea technology, applied in the preservation of human or animal bodies, chemicals for biological control, disinfectants, etc., can solve the problems of expensive, unsuitable, etc., to promote repair, stimulate proliferation, and promote adhesion The effect of growth

Inactive Publication Date: 2013-07-10
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the most commonly used Dexsol and Optisol in the world have significant functions in maintaining the activity of corneal endothelial cells, they are expensive and not suitable for my country's national conditions.

Method used

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  • Cornea mid-term preserving fluid and preparation method thereof
  • Cornea mid-term preserving fluid and preparation method thereof
  • Cornea mid-term preserving fluid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of medium-term preservation solution of cornea

[0045] Taking the configuration of 1000ml as an example, the formula is as follows:

[0046]

[0047]

[0048] (1) Dissolve the low-volume basal medium, tissue culture medium 199, low-molecular-weight dextran, chondroitin sulfate, sodium pyruvate, and HEPES buffer salt in 950 ml of double-distilled water, and use a filter to filter and sterilize , put it into a sterile container, and prepare it as a base solution.

[0049] (3) Add calf serum, non-essential amino acids, gentamicin sulfate and basic fibroblast growth factor in the above ratio to the base solution to make the whole solution, and obtain the mid-term corneal preservation solution.

[0050] The medium-term corneal preservation solution was divided into sterile 20 ml glass bottles, sealed and stored in a refrigerator at 4°C for use.

Embodiment 2

[0051] Embodiment 2, the preparation of medium-term preservation solution of cornea

[0052] Taking the configuration of 1000ml as an example, the formula is as follows:

[0053]

[0054] (1) Dissolve the low-volume basal medium, tissue culture medium 199, low-molecular-weight dextran, chondroitin sulfate, sodium pyruvate, and HEPES buffer salt in 950 ml of double-distilled water, and use a filter to filter and sterilize , put it into a sterile container, and prepare it as a base solution.

[0055] (2) Add calf serum, non-essential amino acids, gentamycin sulfate and basic fibroblast growth factor in the above ratio to the base solution to make the whole solution, and obtain the mid-term corneal preservation solution.

[0056] The medium-term corneal preservation solution was divided into sterile 20 ml glass bottles, sealed and stored in a refrigerator at 4°C for use.

Embodiment 3

[0057] Example 3, corneal endothelial cell density and corneal endothelial cell death rate experiment after corneal mid-term preservation

[0058]A total of 45 cases of fresh human corneal tissues (donors who died of various causes, collected within 6 hours) were collected from Peking University Third Hospital from January 2010 to January 2011, and placed in corneal preservation bottles for eye banks. Each corneal material used 20 milliliters of the medium-term corneal preservation solution prepared in Example 1, sealed the bottle with a parafilm, and stored it in a refrigerator at 4°C. The corneal material was monitored using the HAI EB-3000xyz eye bank endothelial microscope, and the detection time points were 0 day, 3 days, 7 days and 14 days. At each time point, the remaining corneal rings after penetrating keratoplasty were taken, and 2.5g / L trypan blue solution was dropped on the endothelial surface. After 1-2 minutes, the dye was washed with normal saline, and then 2g / L...

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Abstract

The invention discloses cornea mid-term preserving fluid and a preparation method thereof. Based on 1000mL, the mid-term preserving fluid consists of the following ingredients: 4g to 5g of minimum essential medium, 4g to 5g of tissue culture medium 199, 10g to 20g of low-molecular-weight dextranum, 20g to 30g of chondroitin sulfate, 100mg to 150mg of sodium pyruvate, 3g to 4g of HEPES, 20ml to 40ml of calf serum, 10ml to 20ml of non-essential amino acid, 1ml to 2ml of gentamicin sulphate, 100 microliters to 200 microliters of basic fibroblast growth factor and the balance of double distilled water. The cornea mid-term preserving fluid can promote the repair of damaged cells on corneal endothelium and the reconstruction of cellular morphology, can adjust the expression of different integrins on the surface of a cell membrane, can promote the adhesion growth of the cell and can stimulate the proliferation of the endothelial cells, and an effect for reducing the cell apoptosis and maintaining the morphology and activity of the cells can be realized for the mid-term preserved cornea.

Description

technical field [0001] The invention relates to a mid-term corneal preservation solution and a preparation method thereof. Background technique [0002] Corneal preservation is one of the important technologies of eye banks. It is divided into active and inactive preservation. Active preservation is divided into short-term, medium-term and long-term preservation according to the preservation time of corneal endothelial cell activity. Short-term storage is to store the whole eyeball in a wet room at 4°C for 24 hours after disinfection and aseptic treatment. For medium-term preservation, corneal tissue nutrient solution is generally used for preservation at 4°C. At present, Dexsol, Optisol and Optisol-GS are most commonly used in the world, and the preservation time can reach 14 days. Long-term preservation methods include organ culture and ultra-low temperature cryopreservation. In 1965, Capella and Kaufman used the cryopreservation method to effectively extend the preserva...

Claims

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Application Information

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IPC IPC(8): A01N1/02A01P1/00
Inventor 靳瑛洪晶马志中
Owner PEKING UNIV THIRD HOSPITAL
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