A kind of medium-term preservation solution of cornea and preparation method thereof
A technology for preserving liquid and cornea, applied in the preservation of human or animal body, chemicals for biological control, disinfectants, etc., can solve the problems of high price and incompatibility, and achieve the goal of promoting repair, stimulating proliferation and reducing cells. effect of apoptosis
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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 into a sterile container, and prepare the 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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