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Rupid red blood cell freezing and defreezing method, and freeze protective liquid thereof

A technology of red blood cells and thawing solution, which is applied in the direction of biochemical equipment and methods, applications, and preservation of human or animal bodies. It can solve the problems of tedious and time-consuming cryopreservation of reagent red blood cells, achieve convenient preservation and application, and save experiments. Time, the effect of improving the utilization rate

Inactive Publication Date: 2007-03-28
SHANGHAI BLOOD CENT
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Problems solved by technology

[0013] One of the purposes of the present invention is to provide a new method for long-term cryopreservation of red blood cells, which solves the problems of cumbersome and time-consuming red blood cell cryopreservation techniques for existing reagents

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  • Rupid red blood cell freezing and defreezing method, and freeze protective liquid thereof

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Embodiment Construction

[0027] The following experiment compares different concentrations of dextran and citric acid buffer and classic cryoprotective solution. The methods are both rapid liquid nitrogen cryopreservation and rapid thawing. After washing and packing fresh red blood cells, take 0.5ml of each to prepare a 50% red blood cell suspension with different cryoprotective solutions, mix and drop into liquid nitrogen and place for 2 minutes Around, take out the frozen red blood cell pellets, immediately put them into the corresponding thawing solution, pipetting and mixing with a pipette, wait until the red blood cells are completely melted, centrifuge for the first time, take the supernatant for testing; after all the supernatant, add physiological saline, mix Homogenize, place at room temperature for 1 hour, centrifuge for the second time, and take the supernatant for testing. Exhaust the supernatant of the second centrifugation, add saline to 1 ml of the remaining packed cells, and use the COULTE...

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Abstract

The invention relates to experiment using reagent erythrocyte long term storage technique. The erythrocyte icing protecting solution / thawing solution is formed by isotonic solution and macromolecular substance with biocompatibility. The osmotic pressure of the former is 250-400mOsm / L; the molecular weight of the latter is 20000-400000; and its density is 15-40%. The invention utilizes isotonic protecting solution, adds into macromolecular substance colloidal medium to form erythrocyte icing protecting solution to make it not occur hyper-osmotic state inside and outside. And in thawing process, it uses the solution likes with the icing protecting solution to thaw the erythrocyte, washes by physiological saline to remove the macromolecular substance to gain the erythrocyte brine suspended solution. Using in the correlation testing, the whole icing and thawing procedure is about 15min; and the yield of the erythrocyte can reach over 80%.

Description

Technical field [0001] The invention relates to a long-term preservation technology of reagent red blood cells for experiments. Background technique [0002] In the prior art, the red blood cells are suspended in a high osmotic pressure cryopreservation protection solution, in which the main effective component is glycerol or glucose. [0003] For example: Meryman solution: 57% glycerin, 3% sodium lactate, 0.2% Na2HPO4, 0.03% KCl. [0004] Huggins solution: 79% glycerin, 8% glucose, 1% fructose, 0.3% EDTA-2Na. [0005] Krijnen solution: 35% glycerol, 2.9% sorbitol, 0.63% NaCl. [0006] Valeri solution: 35% glycerol, 2.88% mannitol, 0.65% NaCl. [0007] Its working principle is to allow high osmotic pressure glycerol to penetrate into the red blood cells and replace the water in it, thereby reducing the formation of ice crystals inside and outside the red blood cells during the freezing process, and protecting the red blood cells from being destroyed. In the process of thawing red...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08A01N1/02
Inventor 向东
Owner SHANGHAI BLOOD CENT
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