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Composite cryopreservation solution and preparation method and application thereof

A technology of cryopreservation solution and inhibitor, which is applied in the field of compound cryopreservation solution and its preparation, can solve the problems of poor performance and inability to meet the clinical needs of adipose stem cell gel, and achieve the purpose of reducing solute damage, promoting the transmission of ions through the membrane, The effect of freezing point depression

Active Publication Date: 2019-12-10
梵美国际再生医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the general cryopreservation solution has poor performance and cannot meet the clinical needs of adipose-derived stem cell gel.

Method used

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  • Composite cryopreservation solution and preparation method and application thereof
  • Composite cryopreservation solution and preparation method and application thereof
  • Composite cryopreservation solution and preparation method and application thereof

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preparation example Construction

[0036] The present invention also provides a preparation method of the above-mentioned compound cryopreservation solution, which comprises the following steps: mixing polyvinylpyrrolidone, trehalose and glycerin with a solvent, thoroughly mixing, performing filter sterilization, and then adding the remaining components to obtain the compound Freezing solution.

[0037] The present invention also provides an application of the above-mentioned composite cryopreservation solution in cryopreserving adipose stem cell gel.

[0038] Specifically, mix fresh sterile adipose-derived stem cell gel and compound cryopreservation solution according to the volume ratio of 1:1, put them into cryopreservation tubes and let them stand at 2-6°C for 5-15 minutes, then put them into the cell programmed cooling box and transfer Store at -80°C for 16 to 24 hours, then transfer the cryopreservation tube to liquid nitrogen for long-term storage. When resuscitating, take out the cryopreservation tube ...

Embodiment 1

[0041] Weigh polyvinylpyrrolidone (2.5mmol / L), trehalose (0.25mol / L) and glycerin (2.17mol / L) according to the volume of the compound cryopreservation solution to be prepared and the concentration of each component, and add them to Hank's balanced salt solution In the process, use a desktop vortex oscillator at 3000 rpm to fully mix until the white blocky powder disappears and the solution is clear, then use a microporous filter with a pore size of 0.22 μm to filter and sterilize, and then use micropipetting according to the concentration Take Fasudil Hydrochloride (10 μmol / L) with a gun and add to the solution and mix evenly to obtain a composite cryopreservation solution.

Embodiment 2

[0045] Weigh polyvinylpyrrolidone (2.5mmol / L), trehalose (0.25mol / L) and glycerin (2.17mol / L) according to the volume of the compound cryopreservation solution to be prepared and the concentration of each component, and add them to Hank's balanced salt solution In the process, use a desktop vortex oscillator at 3000 rpm to fully mix until the white blocky powder disappears and the solution is clear, then use a microporous filter with a pore size of 0.22 μm to filter and sterilize, and then use micropipetting according to the concentration Add glutathione (5 mmol / L) and fasudil hydrochloride (10 μmol / L) to the solution with a gun and mix well to obtain a composite cryopreservation solution.

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Abstract

The invention relates to a composite cryopreservation solution and a preparation method and an application thereof. The composite cryopreservation solution is mainly composed of the following components: polyvinylpyrrolidone, trehalose, glycerin, a Rock inhibitor and a solvent. The composite cryopreservation solution is composed of polyvinylpyrrolidone, trehalose, glycerin, the Rock inhibitor andthe like as the main components without addition of dimethylsulfoxide, so the risk of dimethylsulfoxide cytotoxicity is eliminated, the composite cryopreservation solution can be used for long-term oreven permanent preservation of adipose stem cell gel in ultra-low temperature, so as to provide an active, safe and effective self filler for future fat transplantation. After 6 months of cryopreservation, the adipose stem cell gel cryopreserved with the compound cryopreservation solution is transplanted into an animal body, and after 3 months, the adipose stem cell gel still shows good retentionrate and tissue morphology, and no complication or adverse reaction appears.

Description

technical field [0001] The invention relates to the field of cell cryopreservation, in particular to a compound cryopreservation solution and its preparation method and application. Background technique [0002] Adipose stem cell gel (SVF-gel, Stromal Vascular Fraction-gel) is a concentrated product of human adipose tissue processed by pure physical methods after degreasing, rich in adipose-derived stem cells and other functional stromal vascular component (SVF) cells, because of its texture As delicate as a colloid, it can be injected through a fine needle, named as adipose stem cell glue. Adipose-derived stem cell gel can be used as a tissue filler similar to stem cell enrichment, and has great potential for clinical treatment. Due to the removal of lipid components of fat, the transplant survival rate of SVF-gel is about 30%-50% higher than that of traditional fat. In addition, due to its multiple enrichment of adipose-derived stem cells, it has shown good application p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 鲁峰冯晶玮姚尧董自清阳兵
Owner 梵美国际再生医学科技有限公司
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