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Production method and formula of lymphocyte separation medium

A lymphocyte and separation liquid technology, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve the problems of cell toxicity, affect cell activity, affect ion permeable membrane, etc., and achieve low production costs. , the effect is good, the production method is simple

Inactive Publication Date: 2020-01-21
无锡华精生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because dextran is obtained from glucose through bacterial fermentation, its endotoxin level is high, which will affect the surface protein activity of lymphocytes, and the separation liquid contains impermeable ion diatrizoate (diatrizoate), Affects the Gibbs-Donnan equilibrium of the ion-permeable membrane (Gibbs-Donnan membrane equilibrium)
Therefore, the traditional lymphocyte separation liquid is toxic to the cells, and the state of the separated cells is not good, which affects the activity of the cells, which is not conducive to the subsequent cultivation of the extracted cells and the application of clinical treatment.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The formula of a kind of lymphocyte separation liquid of the present embodiment specifically includes the following components: sucrose, meglumine, diatrizoic acid and chlorpromaine, and the quality of each component is: 8 kg of sucrose, 3 kg of meglumine, diatrizoic acid 3kg and chlorpromamine 8kg.

[0019] A production method of lymphocyte separation fluid, comprising the following steps:

[0020] 1). Polysucrose is synthesized from sucrose and diatrizoic acid;

[0021] 2). Meglumine and chlorpromaine are prepared into a solution;

[0022] 3). Control the pH value of step 1 and step 2 at 6.5;

[0023] 4). Wrap the solution prepared in step 2 with dialysis paper, put it in a pure water tank and cultivate it for one week, take it out after one week and check that it cannot contain chloride ions.

[0024] 5). Concentrate the solution cultivated in step 4 in a vacuum, and prepare a solution with a density of 1.077 to produce a lymphocyte fluid.

Embodiment 2

[0026] The formula of a lymphocyte separation liquid in this embodiment specifically includes the following components: sucrose, meglumine, diatrizoic acid and chlorpromaine, and the quality of each component is: 10 kg of sucrose, 3.2 kg of meglumine, diatrizoate Acid 5kg and chlorpromamine 9kg.

[0027] A production method of lymphocyte separation fluid, comprising the following steps:

[0028] 1). Polysucrose is synthesized from sucrose and diatrizoic acid;

[0029] 2). Meglumine and chlorpromaine are prepared into a solution;

[0030] 3). Control the pH value of step 1 and step 2 between 6.5;

[0031] 4). Wrap the solution prepared in step 2 with dialysis paper, put it in a pure water tank and cultivate it for one week, take it out after one week and check that it cannot contain chloride ions.

[0032] 5). Concentrate the solution cultivated in step 4 in a vacuum, and prepare a solution with a density of 1.078 to produce a lymphocyte fluid.

Embodiment 3

[0034] The formula of a lymphocyte separation liquid in this embodiment specifically includes the following components: sucrose, meglumine, diatrizoic acid and chlorpromaine, and the quality of each component is: 12 kg of sucrose, 3.5 kg of meglumine, diatrizoate Acid 6kg and chlorpromaline 10kg.

[0035] A production method of lymphocyte separation fluid, comprising the following steps:

[0036] 1). Polysucrose is synthesized from sucrose and diatrizoic acid;

[0037] 2). Meglumine and chlorpromaine are prepared into a solution;

[0038] 3). Control the pH value of step 1 and step 2 at 7.5;

[0039] 4). Wrap the solution prepared in step 2 with dialysis paper, put it in a pure water tank and cultivate it for one week, take it out after one week and check that it cannot contain chloride ions.

[0040] 5). Concentrate the solution cultivated in step 4 in a vacuum, and prepare a solution with a density of 1.078 to produce a lymphocyte fluid.

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Abstract

The invention belongs to the technical field of lymphocyte separation, and particularly relates to a production method and formula of a lymphocyte separation medium. According to the formula of the lymphocyte separation medium, the lymphocyte separation medium is specifically prepared from the following components by mass: 8-12kg of sucrose, 3-3.5kg of meglumine, 3-6kg of amidotrizoic acid and 8-10kg of chloropropylamine. The production method and formula of the lymphocyte separation medium have the beneficial effects that the production method is simple, the extracted cells are twice more than a conventional method, the production cost is low, the effect is good, and the production method and formula are in the leading position in the industry.

Description

technical field [0001] The invention belongs to the technical field of lymphocyte separation, and in particular relates to a production method and formula of a lymphocyte separation liquid. Background technique [0002] The collection of pure lymphocyte populations is based on the ability of monocytes to actively adhere to glass, plastic, nylon wool, cotton fiber or dextran gel at 37°C and in the presence of Ca2+, and remove them from the mononuclear cell suspension. monocytes to obtain a pure lymphocyte population. The main methods are: ①adhesive and adherent method; ②adsorption column filtration method; ③magnet attraction method, and the domestic lymphocyte separation liquid has a history of more than 20 years. The density is different. After centrifugation in the separation medium with a certain density, various types of cells are separated. However, because dextran is obtained from glucose through bacterial fermentation, its endotoxin level is high, which will affect t...

Claims

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Application Information

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IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/00
Inventor 丁秀亮宋桂芳
Owner 无锡华精生物科技有限公司
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