A kind of separation method of blood cell separation tube and mononuclear cells

A technology for separating tubes and blood cells, which is applied in the field of biomedicine, can solve the problems of density gradient centrifugation, cumbersome sampling operations, etc., and achieve the effects of clear superposition interface, reducing errors, and simplifying the operation process

Active Publication Date: 2019-10-15
BEIJING SHENGPU BODA BIOTECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0023] In order to solve the density gradient centrifugation method adding and loading, the complicated problem of sampling operation, the purpose of the present invention is:

Method used

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  • A kind of separation method of blood cell separation tube and mononuclear cells
  • A kind of separation method of blood cell separation tube and mononuclear cells
  • A kind of separation method of blood cell separation tube and mononuclear cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0058] Example 1 An application of the present invention is that the target cells can be separated according to a simplified operation method without using a sample loading slide. figure 2 It is a schematic diagram of this scheme. In this implementation, the pre-separated blood sample can be poured directly into the separation tube. During the process of pouring the blood sample into the separation tube, the separation diaphragm will prevent the sample from mixing with the separation liquid. Select 2000 rpm After centrifugation, the red blood cells are deposited at the bottom of the separation tube through the separation diaphragm, and the mononuclear cells are enriched between the cell separation liquid and the plasma layer above the separation diaphragm. When sampling, pour the liquid above the separation diaphragm directly into a new test tube.

[0059] Table 1 The concentration, purity and viability of lymphocytes separated by two methods

[0060]

[0061] Table 1 is ...

example 2

[0062] Example 2 Another application of the present invention is: combining a sample loading slide with a common centrifuge tube can simplify the sample loading process. image 3 It is a schematic diagram of this program. In this embodiment, when adding the sample, the blood sample is also directly poured into the centrifuge tube, and then the blood is superimposed on the surface of the separation liquid by means of the sample loading slide plate, and the centrifugal speed of 2000 rpm is selected. After centrifugation for 25 minutes, red blood cells were deposited at the bottom of the centrifuge tube, and the target cell layer was aspirated with a Pasteur pipette to complete the separation of mononuclear cells.

[0063] Table 2 Comparison of lymphocyte concentration, purity and viability of two conventional separation methods

[0064]

[0065] Table 2 is the statistical results of mononuclear cell concentration, cell purity and cell viability separated according to this pro...

example 3

[0066] Example 3 Due to the flow-limiting effect of the separation diaphragm, under the action of centrifugal force, the flow velocity of the liquid through the separation diaphragm will change. In order to reduce the impact of the change of flow velocity on the separation effect, the present invention stabilizes the flow coefficient by increasing the number of small holes on the separation diaphragm. At the same time, appropriately increase the amount of separation liquid, so that the added separation liquid is 1-2 cm higher than the separation diaphragm, and the separation liquid above the separation diaphragm plays a role in stabilizing the flow rate, thereby achieving the best separation effect. Figure 4It is a schematic diagram of this scheme. In this implementation method, the sample is superimposed on the separation liquid with the help of the sample loading slide, and then the sample loading slide is moved upward, and the centrifugal speed of 2000 rpm is selected for 25...

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Abstract

The invention provides a blood cell separation tube and a method for separating a mononuclear cell by using the blood cell separation tube. The blood cell separation tube consists of a separation tube body, a rotary cover as well as a cell separation membrane and a sample adding slide plate (an auxiliary sampling device) which are arranged in the tube body. According to the method, a conventional complicated sample feeding mode is changed to an easily operated mode by changing sample-adding and sampling modes on the basis of a principle that the cell is separated through cell density gradient centrifugation. According to the blood cell separation tube and the method provided by the invention, a cell separation process can be more simplified and the operation can be easier in clinical medical examination and cell biology research.

Description

technical field [0001] The invention relates to cell separation technology in the field of biomedicine. The blood cell separation tube and the method for separating mononuclear cells provided by the present invention are based on the principle of cell density gradient centrifugation, and change the traditional and complicated sampling mode into an easy-to-operate mode by changing the sampling and sampling methods. Background technique [0002] In clinical medical testing, the examination of specific cell functions and biological characteristics often involves cell separation techniques. Isolation of mononuclear cells from peripheral blood and various body fluids (pleural effusion, ascites, pericardial effusion, etc.) is performed in vitro. An important pretreatment step for cellular immunology research and cytological experiments, the quality and quantity of the isolated target cells is an important link to ensure the reliability of subsequent experiments. [0003] Peripher...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/24C12M1/12C12N5/078
Inventor 宋雪程月红
Owner BEIJING SHENGPU BODA BIOTECH
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