A kind of preparation method and application of cell, tissue or organ cold preservation solution

A preservation method and technology of preservation solution, applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve the problems of unfavorable preservation solution improvement, lack of protection mechanism guidance of organ preservation solution, failure of corneal transplantation operation, etc., to achieve survival Better, guaranteed quality, increased corneal permeability

Active Publication Date: 2022-05-27
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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  • A kind of preparation method and application of cell, tissue or organ cold preservation solution
  • A kind of preparation method and application of cell, tissue or organ cold preservation solution
  • A kind of preparation method and application of cell, tissue or organ cold preservation solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Activation of FOXO1 enables younger corneas to have better cold adaptation

[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 compared with the cornea of ​​6-month-old rats, the corneal endothelial cells of 4-week-old rats died less, indicating that the corneal tissue of young individuals has better cold adaptation ability.

[0071] The inventor's preliminary research results show that the entry of FOXO1 protein into the nucleus is an important mechanism for cold adaptation in hibernating animals (for details, see patent WO 2019 / 040359A1, and Jingxing Ou et al.'s document "iPSCs from aHibernator Provide a Platform for Studying Cold Adaptation and Its Potential Medical Applications" , published on: 2018.05.03, DOI: 10.1016 / j.cell.2018.03.010). By immunofluorescence staining, we found that ( figure 2 ), FOXO1 was in t...

Embodiment 2C6

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

[0073] The donor sources of corneal transplantation are mainly adults and the elderly. Is it possible to activate FOXO1 to enable adult corneas to acquire the cold adaptation ability of young individual corneas? In this regard, the inventors successfully screened two drugs, C6 ceramide (C6 for short) and Apigenin.

[0074] The inventors 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 injection of C6, the Mouse corneal endothelial cells can activate FOXO1 into the nucleus and the nuclear entry signal is enhanced after cold stimulation ( image 3 ). However, injection of Apigenin did not cause FOXO1 to enter the nucleus. After cold stimulation, the nuclear entry signal activated FOXO1 into the nucleus, and the weak nucle...

Embodiment 3

[0076] Embodiment 3 Novel corneal preservation solution formula

[0077] According to the metabolic characteristics during hibernation, the inventors mainly adjusted polar neutral amino acids, basic amino acids and water-soluble vitamins, and adjusted the components of the existing preservation solution 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 cold preservation solution for cells, tissues or organs. The present invention is based on the cold adaptation mechanism and patent WO2019 / 040359 A1 discovered by the inventor in the previous research on hibernating model animals, and improves the cold preservation effect of donor cells, tissues or organs in vitro by regulating the entry of FOXO1 into the nucleus with drugs. Taking corneal cold preservation as an example, under the cold preservation strategy of the new corneal preservation solution of the present invention plus C6 ceramide, Apigenin and Pi, the survival of corneal endothelial cells is better, the mortality rate is only 2.5%, and the corneal permeability is increased. This guarantees the quality of corneal cryopreservation and provides more optional corneal donors for successful transplantation. The cold adaptation to hibernation is effective in different tissues and species, which suggests that we can improve and develop preservation solutions for different species and organs based on this.

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, organ transplantation techniques have been continuously enriched, the range of organ transplantation donors has continued to expand, and the survival rate of organ transplant recipients has continued to improve. However, existing organ donations are far from meeting the huge demand for transplant donors, and it is particularly important to make good use of limited organ resources. The rapid development of organ static cold storage (SCS) technology for organ protection solution has further freed organ preservation from time and geographical constraints. Since then, the development of organ protection has entered a new era, ...

Claims

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Application Information

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Patent Type & Authority Patents(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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