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Low-temperature preservation method and rewarming method of blood vessels

A technology for cryopreservation and blood vessels, which is applied in the field of cryogenic medicine to achieve the effects of inhibiting the formation of ice crystals, reducing osmotic pressure differences, and simple operation

Active Publication Date: 2020-03-20
银丰低温医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned prior art, in order to solve the problem of temperature difference between inside and outside of large-volume tissue and organ cooling, the present invention provides a blood vessel cryopreservation method and rewarming method, and also provides a blood vessel cryoprotectant

Method used

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  • Low-temperature preservation method and rewarming method of blood vessels
  • Low-temperature preservation method and rewarming method of blood vessels
  • Low-temperature preservation method and rewarming method of blood vessels

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The screening of embodiment 1 cryoprotectants

[0033] Isolate and obtain fresh vascular tissue, wash the inside and outside of the blood vessel with cleaning solution (RPMI1640 medium containing 10% fetal bovine serum) at 20-25°C (remove blood residue); %, 50%, 75%, 100%) perfuse the cryoprotectant into the inside of the blood vessel (to ensure that the inside of the blood vessel contains sufficient cryoprotectant), and then soak the blood vessel in the cryoprotectant of the same concentration gradient for 3min, the final concentration After perfusion, place in a cryopreservation tube or a cryopreservation bag (depending on the length and thickness of the blood vessel), seal the seal, and carry out cooling. Stored in nitrogen; the vitrified solution was directly put into liquid nitrogen for cryopreservation.

[0034] Rewarming was carried out by means of water bath rewarming: the cryopreserved blood vessel samples were taken out of the liquid nitrogen, the outer sleeve ...

Embodiment 2

[0046] Embodiment 2 Determination of programmatic cooling mode cooling rate

[0047] For slightly larger tissues such as blood vessels, the traditional vitrification cooling method, due to its more complex cryoprotectant composition, and the uneven heat transfer inside and outside the blood vessel during the rapid cooling process of vitrification, causes uneven cooling inside and outside the blood vessel. To a certain extent, it increases the possibility of vascular rupture and spatial structure damage. The introduction of programmed cooling can largely solve a series of problems caused by vitrification cooling. On the basis of determining the optimal cryoprotectant in Example 1, explore different ways of cooling, especially determine the difference between programmed cooling and vitrification.

[0048] Considering the composition and DSC analysis of the cryoprotectant screened in Example 1, in the traditional blood vessel cryopreservation process, in the temperature range of...

Embodiment 3

[0052] Embodiment 3 Magnetic-thermal rewarming of frozen blood vessels

[0053] For the traditional cryopreservation methods of cells and tissues, especially for the preservation of a small number of cells, vitrification is a good way. When tissues and organs are preserved, the rewarming process is more likely to cause damage to the tissue structure.

[0054] On the basis of Example 1 and Example 2, it is determined that under the situation of traditional water bath rewarming, the cryoprotectant of the present invention is used to drop to -40°C at -1°C / min, and to -5°C / min to -40°C. -100°C, and then put it into liquid nitrogen to cool down the blood vessel tissue, and store it in liquid nitrogen for a long time. In the rewarming process of blood vessels, there will be more or less uneven rewarming inside and outside. The introduction of nanoparticles can better achieve uniform rewarming.

[0055] Magnetic nanoparticles, the magnetic nanoparticles are ferric oxide (Fe 3 o 4...

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Abstract

The invention discloses a low-temperature preservation method of blood vessels. The low-temperature preservation method comprises the following steps: separating to obtain fresh blood vessel tissues,pouring a cryoprotectant into the blood vessels, putting the blood vessels into a cryoprotectant or a cryoprotectant containing magnetic nanoparticles, putting an obtained mixture into a cryopreservation tube or a cryopreservation bag, sealing, and carrying out programmed cooling; wherein the cryoprotectant comprises the following components 5%-8% of hydroxyethyl starch, 10%-15% of dimethyl sulfoxide, 0.01%-0.02% of a surfactant and the balance a culture medium. The invention also discloses a rewarming method of blood vessels. The rewarming method of blood vessels comprises the following steps: taking out a blood vessel sample cryopreserved by using the cryoprotectant containing the magnetic nanoparticles from liquid nitrogen, placing the blood vessel sample in an alternating magnetic field area, and carrying out magneto-thermal rewarming under the conditions that the magnetic field frequency is 764.8 kHz, the magnetic field intensity is 250-300Gs and the time is 40-60s. According to the blood vessel cryopreservation method, blood vessel breakage or space structure damage caused by different cooling rates of the temperature inside and outside the blood vessels is avoided, the mechanical property of the blood vessels is protected from being affected as far as possible, operation is easy, and efficiency is high.

Description

technical field [0001] The invention relates to a low-temperature preservation method and a rewarming method for blood vessels, in particular to a deep-low temperature preservation solution for blood vessel tissue, a cooling method and a rewarming method, and belongs to the field of low-temperature medicine. Background technique [0002] Vascular tissue disease is a clinically common disease, such as vasculitis, phlebitis and arteriosclerosis. Under certain conditions, recanalization of blood vessels is required, and vascular grafting is required when necessary. At present, about tens of millions of people in the world need tissue or organ transplantation to treat cardiovascular diseases every year, and only 15% of them can be cured by repairing their own blood vessels, while the vast majority of patients can only be cured by transplanting other people's normal blood vessels. Blood vessels can cure diseases and restore health. The development of vascular surgery in recent ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/021A01N1/0221A01N1/0284A01N1/0294
Inventor 刘峰董一霖臧传宝胥义刘琇
Owner 银丰低温医学科技有限公司
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