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A blood vessel ice crystal-free cryopreservation solution and blood vessel preservation method

A technology of cryopreservation solution and preservation method, which is applied in the field of blood vessel ice crystal-free cryopreservation solution and blood vessel preservation, and can solve the problems of decreased secretion of vasodilation substances such as NO, long-term patency rate affected by luminal stenosis, and decreased vasoconstriction and relaxation function. , to achieve the effect of improving the long-term patency rate, improving the long-term patency rate, optimizing the cooling rate and resuscitating heating rate

Active Publication Date: 2020-10-30
江苏科瑞百奥生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Damage to vascular smooth muscle cells can greatly reduce vasoconstriction and relaxation
After the endothelial cells of the vascular intima are damaged and peeled off, the vascular intima will thicken, and the smooth muscle cells will proliferate and migrate to the subintima to develop into the main cells, causing luminal stenosis and affecting the long-term patency rate and other adverse consequences; at the same time, the endothelial cells The damage of the blood vessels will reduce the secretion of NO and other vasodilation substances, resulting in vascular contracture; in addition, the damage of vascular endothelial cells will reduce the secretion of heparin, and the less heparin production will easily lead to the formation of thrombus and affect the long-term patency rate.

Method used

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  • A blood vessel ice crystal-free cryopreservation solution and blood vessel preservation method
  • A blood vessel ice crystal-free cryopreservation solution and blood vessel preservation method
  • A blood vessel ice crystal-free cryopreservation solution and blood vessel preservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1. Take 3.10mol glycerin, 3.70mol formamide, 0.5mol polyvinylpyrrolidone, 0.5mg / l nitroglycerin, 30mg / l CGS26303, 40μmol / l Z-DEVD, mix well and add Euro-Collin solution to make the volume to 1L.

[0047] 2. Take an appropriate amount of the solution in step 1, and cool it to 2-4°C. Add the cooled solution to the human vascular tissue to be frozen in a 6-step method at 2-4°C, and complete the introduction within 90 minutes (at 2-4°C, step by step in 15 minutes, follow the 6 steps Incrementally add ice-free cryopreservation solution to the vascular tissue, 15 minutes each time).

[0048] 3. The human blood vessel tissue introduced into the cryopreservation solution in the previous step was cooled to -100°C at a rate of -43°C / min, and then the sample was cooled to -135°C at a rate of -3±0.2°C / min, and kept at this temperature Store at least 24h under low temperature; after that, the sample can be stored at -135℃ for a long time.

[0049] 4. Take out the blood vessel sample to b...

Embodiment 2

[0052] 1. Take 3.10mol glycerin, 3.70mol formamide, 0.5mol polyvinylpyrrolidone, 0.5mg / l nitroglycerin, mix well and add Euro-Collin solution to make the volume to 1L.

[0053] 2. Take an appropriate amount of the solution in step 1, and cool it to 2-4°C. Add the cooled solution to the human vascular tissue to be frozen in a 6-step method at 2-4°C, and complete the introduction within 90 minutes (at 2-4°C, step by step in 15 minutes, follow the 6 steps Incrementally add ice-free cryopreservation solution to the vascular tissue, 15 minutes each time).

[0054] 3. Cool the human blood vessel tissue from the previous step to -100°C at a rate of -43°C / min, then cool the sample to -135°C at a rate of -3±0.2°C / min, and store it at this temperature for at least 24h ; After that, the sample can be stored at -135℃ for a long time.

[0055] 4. Take out the blood vessel sample to be resuscitated, restore to -70°C at a rate of 30±0.2°C / min, and then restore to 4°C at a rate of 225±15°C / min.

[...

Embodiment 3

[0058] 1. Take 3.10mol glycerin, 3.70mol formamide, 0.5mol polyvinylpyrrolidone, 30mg / l CGS26303 and mix well and add Euro-Collin solution to make the volume to 1L.

[0059] 2. Take an appropriate amount of the solution in step 1, and cool it to 2-4°C. Add the cooled solution to the human vascular tissue to be frozen in a 6-step method at 2-4°C, and complete the introduction within 90 minutes (at 2-4°C, step by step in 15 minutes, follow the 6 steps Incrementally add ice-free cryopreservation solution to the vascular tissue, 15 minutes each time).

[0060] 3. The human blood vessel tissue introduced into the cryopreservation solution in the previous step was cooled to -100°C at a rate of -43°C / min, and then the sample was cooled to -135°C at a rate of -3±0.2°C / min, and kept at this temperature Store at least 24h under low temperature; after that, the sample can be stored at -135℃ for a long time.

[0061] 4. Take out the blood vessel sample to be resuscitated, restore to -70°C at a...

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Abstract

The invention discloses a blood vessel ice-crystal-free cryopreservation solution and a blood vessel preservation method. The blood vessel ice-crystal-free cryopreservation solution comprises 1.0-6.0mol / L of glycerol, 1.0-7.0 mol / L of formamide, 0.1-2.5 mol / L of polyvinylpyrrolidone and a component X; wherein the component X is one or more of a component A, a component B and a component C; wherein the component A is an NO donor, the component B is an endothelin inhibitor or an endothelin antagonist, and the component C is an anti-apoptotic agent. According to the preserving fluid disclosed bythe invention, the preserving effect on the vascular function is improved, especially for vascular smooth muscle cells and endothelial cells which are most easily subjected to cryopreservation injury; the vascular smooth muscle cell and the endothelial cell can be effectively protected, the function of effectively protecting the vascular smooth muscle cell and the endothelial cell is achieved; inaddition, the long-term patency rate after vascular transplantation is increased, and the success rate of clinical vascular transplantation is greatly guaranteed.

Description

Technical field [0001] The invention belongs to the field of biomedical engineering, and specifically relates to a blood vessel ice crystal-free cryopreservation liquid and a blood vessel preservation method. Background technique [0002] In the past 20 years, the clinical demand for vascular substitutes has increased year by year. The application of allogeneic vascular transplantation can solve various problems such as vascular injury, aneurysm, atherosclerosis, vascular disease complicated by diabetes, various complicated heart diseases, and establishment of hemodialysis access. In addition, in the field of organ transplantation, when the vascular anastomosis between the donor organ and the host does not match, additional vascular substitutes are also needed to complete the vascular reconstruction of the transplanted organ. In the past, it was an absolute contraindication for malignant tumor tissue to invade adjacent key blood vessels. However, with the development of technolo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02C12N5/071
CPCA01N1/0221A01N1/0226A01N1/0284C12N5/0691C12N2523/00
Inventor 宋云庆欧阳溪
Owner 江苏科瑞百奥生物技术有限公司
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