Preparation method and application of cell, tissue or organ cold preservation liquid

A preservation method and technology of preservation solution, applied in the preparation of tissue or organ cold preservation solution, and in the field of cells, can solve the problems of unfavorable preservation solution improvement, lack of protection mechanism guidance of organ preservation solution, failure of corneal transplantation operation, etc., and achieve corneal permeability. Increased sex, guaranteed quality, better survival

Active Publication Date: 2021-04-06
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cornea after such cryopreservation often leads to the failure of corneal transplantation
The 12-day storage period cannot meet the needs of larger eye bank storage, and a better corneal storage strategy is urgently needed to meet
[0005] The current organ preservation solution lacks a clear guidance on the protection mechanism, which is not conducive to the improvement of the preservation solution, and also limits the scope of use of organ preservation strategies
A systematic cold storage theoretical system has not yet been formed at home and abroad, and it is urgent to explore a better cold storage mechanism to improve the current cold storage strategy.

Method used

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  • Preparation method and application of cell, tissue or organ cold preservation liquid
  • Preparation method and application of cell, tissue or organ cold preservation liquid
  • Preparation method and application of cell, tissue or organ cold preservation liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Activation of FOXO1 enables better cold adaptation in young corneas

[0070] The corneas of 4-week-old rats and 6-month-old rats were stored in commercial Optisol GS preservation solution at 4°C for 3 weeks. The result is as figure 1 As shown, it was found that the corneal endothelial cells of 4-week-old rats died less than those of 6-month-old rats, indicating that young individual corneal tissues have better cold adaptation ability.

[0071] The inventor's previous research results show that the entry of FOXO1 protein into the nucleus is an important mechanism for cold adaptation in hibernating model animals (see patent WO 2019 / 040359A1 for details, and Jingxing Ou et al.'s document "iPSCs from a Hibernator Provide a Platform for Studying Cold Adaptation and Its Potential Medical Applications ", public date: 2018.05.03, DOI: 10.1016 / j.cell.2018.03.010). By immunofluorescence staining, we found that ( figure 2 ), FOXO1 was in the nuclear state in 4-week-o...

Embodiment 2C6

[0072] Example 2C6 Ceramide and Apigenin can activate adult rat corneal FOXO1 into nucleus

[0073] The donors for corneal transplantation are mainly adults and the elderly. Whether it is possible to activate FOXO1 to allow adult corneas to acquire the cold adaptation ability of young individual corneas. In this regard, the inventors successfully screened two drugs, C6 ceramide (abbreviated as C6) and Apigenin.

[0074] The inventor injected C6 ceramide (C6, concentration 20 μM / μL, solvent goat serum) and Apigenin (concentration 4 μM / μL, solvent goat serum) into the anterior chamber of the eyeball of 6-month-old rats, and found that after the injection of C6, large Mouse corneal endothelial cells can activate FOXO1 into the nucleus and the nuclear import signal is enhanced after cold stimulation ( image 3 ). However, the injection of Apigenin does not cause FOXO1 to enter the nucleus. After the cold stimulation, the nuclear import signal activates FOXO1 to enter the nucleus...

Embodiment 3

[0076] Embodiment 3 Novel corneal preservation solution formula

[0077] According to the metabolic characteristics during hibernation, the inventors adjusted the components of the existing preservation solution mainly based on polar neutral amino acids, basic amino acids, and water-soluble vitamins, so as to meet the normal physiological needs of cells during cold storage. The composition of the improved new corneal preservation solution is shown in Table 1.

[0078] Table 1 New corneal preservation solution (MCM) formula

[0079]

[0080]

[0081]

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Abstract

The invention relates to the field of preservation of medical materials, in particular to a preparation method and application of a cell, tissue or organ cold preservation solution. According to a cold adaptation mechanism and a patent WO2019/040359 A1 discovered by early hibernation mode animal research of the inventor, extension is performed, and the in-vitro cold preservation effect of donor cells, tissues or organs is improved by regulating and controlling nucleus entry of FOXO1 through a medicine. Taking cornea cold preservation as an example, under the cold preservation strategy that C6 ceramide, Apigenin and Pi are added into the novel cornea preservation solution, corneal endothelial cells survive better, the death rate is only 2.5%, and the corneal permeability is improved. The quality of cornea in-vitro cold preservation is guaranteed, and more selectable cornea donors are provided for successfully carrying out a transplantation operation. The hibernation cold adaptation is based on effectiveness in different tissues and different species, and it is prompted that we can perform improved development of preserving fluid for different species and different organs.

Description

technical field [0001] The invention relates to the field of preservation of medical materials, in particular to a preparation method and application of a cold preservation solution for cells, tissues or organs. Background technique [0002] Since the invention of organ transplantation technology in the 1850s, through continuous innovation and development, the surgical methods of organ transplantation have been continuously enriched, the range of organ transplant donors has been continuously expanded, and the survival rate of organ transplant recipients has been continuously improved. However, existing organ donations are far from meeting the huge demand for transplant donors, so how to make good use of limited organ resources is particularly important. The rapid development of static cold storage (SCS) technology of organ protection liquid has further freed organ preservation from time and geographical constraints. Since then, the development of organ protection has entered...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/021A01N1/0221A01N1/0226
Inventor 区景行金光辉杨扬刘炜葛礼浩
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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