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Peripheral blood lymphocyte separating medium and application thereof

A lymphocyte and peripheral blood technology, applied in the field of peripheral blood lymphocyte separation fluid, can solve the problems of poor peripheral blood lymphocyte separation, high price of diatrizoate, and low peripheral blood content, so as to reduce the phenomenon of mutual dissolution , less platelet contamination, and high activity rate

Pending Publication Date: 2022-04-15
武汉赛维尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

but diatrizoate is expensive
In addition, mice are commonly used experimental animals in the fields of biology and medical research, but the peripheral blood content of mice is small, and the process of blood collection is very easy to cause hemolysis, resulting in poor and ineffective separation of peripheral blood lymphocytes. Stablize
It is also prone to platelet contamination

Method used

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  • Peripheral blood lymphocyte separating medium and application thereof
  • Peripheral blood lymphocyte separating medium and application thereof
  • Peripheral blood lymphocyte separating medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Effect of Polypropylene Pyrrolidone Concentration on Separation of Peripheral Blood Lymphocytes

[0057] 1. Configure mouse peripheral blood lymphocyte separation medium

[0058] The volume ratio of each component of mouse peripheral blood lymphocyte separation fluid is shown in Table 1, and the configuration steps are as follows:

[0059] Accurately weigh each component, dissolve in ultrapure water, stir to dissolve; adjust the pH to 7.2-7.6, and constant volume; adjust the temperature to 20°C, and perform density detection. Sodium diatrizoate is used for density adjustment, and the density is 1.083± 0.001g / mL; after filter sterilization, store at 4°C.

[0060] Table 1 Group 1~6 mouse peripheral blood lymphocyte separation fluid formula

[0061]

[0062] 2. Experimental method

[0063] To test the separation effect of peripheral blood lymphocytes in mice under different concentrations of polypropylene pyrrolidone. Specific steps are as follows:

[006...

Embodiment 2

[0080] Example 2 Effect of sodium diatrizoate and polysucrose content on the separation effect of peripheral blood lymphocytes in mice

[0081] 1. Configure mouse peripheral blood lymphocyte separation medium

[0082] The volume ratio of each component of the mouse peripheral blood lymphocyte separation fluid is shown in Table 4, and the configuration steps are shown in Example 1.

[0083] Table 4 Group 3 and the mouse peripheral blood lymphocyte separation fluid formula of groups 7-9

[0084]

[0085] 2. Experimental method

[0086] According to the experimental results of Example 1, it is found that the concentration of polypropylene pyrrolidone is preferably between 5 and 10 mg / mL, and the concentration of polypropylene pyrrolidone is controlled between 5 and 10 mg / mL to explore the effects of sodium diatrizoate and polysucrose on mice. Influence on the efficacy of peripheral blood lymphocyte isolation.

[0087] Since the peripheral blood of mice is generally less, an...

Embodiment 3

[0094] Example 3 Peripheral Blood Lymphocyte Separation Fluid Stability

[0095] 1. Configure mouse peripheral blood lymphocyte separation medium

[0096] Configure mouse peripheral blood lymphocyte separation liquid group 10, the content of each component is: 90 mg / mL sodium diatrizoate + 60 mg / mL polysucrose, the volume ratio of each component in group 3 and group 7 is shown in Example 2, and the configuration steps are shown in Example 1.

[0097] 2. Experimental method

[0098] Since the peripheral blood of mice is generally less, another batch of fresh anticoagulated whole blood of mice of the same species and feeding environment was taken to test the stability of the peripheral blood lymphocyte separation medium of mice in groups 3, 7 and 10. Separation was carried out according to the method of Example 1. Take pictures at 0, 5, 10, 15, 30 and 60 minutes to observe the stability of the separation fluid and blood.

[0099] 3. Experimental results

[0100] Such as f...

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Abstract

The invention discloses a peripheral blood lymphocyte separating medium and application thereof. The peripheral blood lymphocyte separating medium is prepared from the following components in parts by mass: 102.8 to 110 parts of sodium diatrizoate, 10 to 30 parts of hydroxyethyl starch, 30 to 45 parts of polysucrose and 2.5 to 20 parts of polypropylene pyrrolidone. While sodium diatrizoate is adopted to replace expensive meglumine diatrizoate, other saccharide or non-saccharide macromolecules are introduced, so that the phenomenon that the separation effect is poor or unstable in the separation process of the mouse peripheral blood stranguria lysozyme is improved. The stability between the separating liquid and the blood can be maintained for a long time, the problem that the blood and the separating liquid are mutually mixed due to too long sample adding time or too large force can be prevented, and the phenomenon that the blood and the separating liquid are mutually dissolved is reduced. And the recovery efficiency, the purity and the motility rate are high, and the platelet pollution is less.

Description

technical field [0001] The invention relates to the technical field of lymphocyte separation, in particular to a peripheral blood lymphocyte separation liquid and an application thereof. Background technique [0002] Peripheral blood lymphocytes (PBL) are a type of white blood cells produced by lymphoid organs, participate in specific immune responses, and play an important role in the immune response mechanism. Detecting the number or proportion, function and strength of peripheral blood lymphocytes with immune response and their components is an important means to judge the level of human cellular immunity. [0003] At present, the separation of peripheral blood lymphocytes is mainly based on the difference in the density of various cells in the blood, using a certain density of separation liquid, and centrifuging to separate various types of cells. The traditional separation medium is a mixture of Ficoll and Meglumine Diatrizoate for the separation of peripheral blood ly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/0783
Inventor 涂佳鹏何小燕李银平
Owner 武汉赛维尔生物科技有限公司
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