Cell separation tube for separating and extracting PBMC (peripheral blood mononuclear cells)
By setting a slope fixation ring in the cell separation tube, the problem of poor separation effect of PBMC in the prior art was solved, and efficient blood sample addition without tilting operation and extraction of high-purity PBMC are achieved, which is suitable for separation experiments of PBMC.
Patent Information
- Application Number
- CN202422206444.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-10
- Publication Date
- 2025-07-18
- Estimated Expiration
- 2034-09-10
AI Technical Summary
In the prior art, the separation effect of PBMC is poor, mainly because the centrifuge tube needs to be tilted and the slow operation is carried out, which can easily lead to the layering interface of the lymphocyte separation fluid being broken, and when the PBMC is absorbed, the plasma and separation fluid are easily mixed into the plasma, affecting the purity and yield.
A cell separation tube was designed to separate and extract PBMCs, which contained a removable connection of the separation tube body and the tube cover. The inner wall was equipped with a fixing ring. The upper and lower surfaces of the fixing ring were sloped, allowing blood samples to be slowly injected along the slope to avoid mixing. After centrifugation, the PBMC was located in the inner circle of the fixing ring, reducing the contact area with the lymphocyte separation fluid and the plasma layer.
It realizes the addition of blood samples without tilting operation, improves the purity and yield of PBMC, ensures efficient separation effect, and is suitable for separation experiments of PBMC.
Smart Images
Figure CN223118455U_ABST
Abstract
Description
Technical Field
[0001] The utility model belongs to the technical field of medical devices and relates to a cell separation tube for separating and extracting PBMCs. Background Art
[0002] PBMC (Peripheral blood mononuclear cell) is a cell with a single nucleus in peripheral blood, including lymphocytes, monocytes, dendritic cells, and other small amounts of hematopoietic stem cells, etc. It is a key barrier for the body to fight against infectious diseases. PBMC is convenient to obtain, and the extraction method is relatively mature, and it has been widely used in various fields of medical research and clinical treatment.
[0003] Since the densities of various blood cells in peripheral blood are different, density gradient centrifugation using lymphocyte separation fluid (Ficoll-Hypaque) is used to make cell populations with a certain specific gravity distribute according to the corresponding density gradient, so as to separate various blood cells, and then extract PBMCs in peripheral blood. During the density gradient centrifugation separation process, first, a certain volume of lymphocyte separation fluid is added to a straight-wall centrifuge tube, and then the blood sample to be separated is very carefully and slowly added to the upper layer of the lymphocyte separation fluid. After density gradient centrifugation, red blood cells and granulocytes sediment at the lower layer of the lymphocyte separation fluid, plasma is located at the upper layer, PBMCs are in a white film shape and are located at the junction of the lymphocyte separation fluid and plasma, and then the middle PBMCs are aspirated with a pipette or a straw.
[0004] The technical difficulty in separating and extracting PBMCs by density gradient centrifugation is that the process of adding the blood sample requires tilting the centrifuge tube and being very careful and slow to prevent the stratification interface of the lymphocyte separation fluid from being broken, resulting in the mixing of blood and the separation fluid. In addition, when aspirating PBMCs in the white film layer, it is very easy to aspirate the upper plasma and the lower separation fluid, or the PBMCs in the white film layer are very easy to be disturbed and mixed with other layers, affecting the separation effect of PBMCs and resulting in a poor separation effect of PBMCs. Summary of the Utility Model
[0005] The purpose of the utility model is to provide a cell separation tube for separating and extracting PBMCs to solve the problem of poor separation effect of existing PBMCs.
[0006] To achieve the above purpose, the utility model adopts the following technical solutions:
[0007] The utility model provides a cell separation tube for separating and extracting PBMCs, which includes a detachable separation tube body and a tube cap. A fixing ring is provided on the inner wall of the separation tube body. The outer diameter of the fixing ring is the same as the inner diameter of the separation tube body, and there is a gap between the bottom surface of the fixing ring and the bottom of the separation tube body; both the upper and lower surfaces of the fixing ring are slopes.
[0008] Preferably, the included angle between the upper surface of the fixing ring and the tube wall of the separation tube body is 30°.
[0009] Preferably, the included angle between the lower surface of the fixing ring and the tube wall of the separation tube body is 75°.
[0010] Preferably, the inner diameter of the fixing ring is 1 / 3 - 1 / 2 of the inner diameter of the separation tube body.
[0011] Preferably, the height at the inner circle of the fixing ring is 1 / 4 - 1 / 3 of the height of the separation tube body, and the height at the outer circle of the fixing ring is 1 / 2 - 2 / 3 of the height of the separation tube body.
[0012] Preferably, the volume between the lower surface of the fixing ring and the bottom of the separation tube body is 15 mL.
[0013] Preferably, the fixing ring is a hollow fixing ring.
[0014] Preferably, the fixing ring is a solid fixing ring.
[0015] Preferably, scales are provided on the outer wall of the separation tube body.
[0016] The utility model has the following beneficial effects:
[0017] (1) Without tilting the separation tube body, the blood sample can be injected into the cell separation tube along the fixing ring with a slope structure, and will not be mixed with the lymphocyte separation liquid due to too fast injection speed.
[0018] (2) The extracted PBMC is located at the inner circle of the fixing ring, and the contact area with the lymphocyte separation liquid and the plasma layer is greatly reduced, which is beneficial to using a pipette or a straw to pipette more high-purity PBMC, reducing the mixing of other components in the blood and the lymphocyte separation liquid into the PBMC, and improving the purity and yield of PBMC.
[0019] (3) The structure of the cell separation tube is reasonable, the separation effect is good, high-purity PBMC can be separated and extracted, and it is suitable for the separation experiment of PBMC. Description of the Drawings
[0020] Figure 1 It is the external structure diagram of the cell separation tube for separating and extracting PBMC provided by the embodiment of the present application;
[0021] Figure 2 It is the three-dimensional structure diagram of the cell separation tube for separating and extracting PBMC provided by the embodiment of the present application;
[0022] Symbol representation:
[0023] 1 - Separation tube body, 2 - Tube cap, 3 - Fixed ring, 4 - Scale. Detailed implementation mode
[0024] Next, the technical solutions in the embodiments of the present utility model will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all of the embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present utility model.
[0025] The embodiment of the present application provides a cell separation tube for separating and extracting PBMC. The cell separation tube includes a detachable separation tube body 1 and a tube cap 2, as shown in the attached Figure 1 , 2 figure. Among them, the separation tube body 1 and the tube cap 2 are connected by threads.
[0026] A fixed ring 3 is provided on the inner wall of the separation tube body 1, and the outer diameter of the fixed ring 3 is the same as the inner diameter of the separation tube body 1. Therefore, the fixed ring 3 is fixed on the entire inner wall of the separation tube body 1. In the embodiment of the present application, the fixed ring 3 is a hollow fixed ring or a solid fixed ring, which can be set according to the actual application scenario.
[0027] A gap is left between the bottom surface of the fixed ring 3 and the bottom of the separation tube body 1 to facilitate the placement of the added lymphocyte separation liquid. In addition, both the upper and lower surfaces of the fixed ring 3 are slopes. Thus, when the lymphocyte separation liquid is added, the lymphocyte separation liquid is located between the bottom surface of the fixed ring 3 and the bottom of the separation tube body 1. When the blood sample to be separated is added, since the upper surface of the fixed ring 3 is a slope, the blood sample can slowly flow into the cell separation tube along the fixed ring 3 with a slope structure without tilting the separation tube body 1, and the stratification interface between the blood sample and the lymphocyte separation liquid will not be damaged, and thus the blood sample will not be mixed with the lymphocyte separation liquid. After centrifugal separation, since the volume of the gap between the bottom surface of the fixed ring 3 and the bottom of the separation tube body 1 is limited, and the inner diameter of the fixed ring 3 is smaller than the inner diameter of the separation tube body 1, the separated and extracted PBMC is located at the inner circle of the fixed ring 3. At this time, the contact area between the PBMC and the lymphocyte separation liquid and the plasma layer is small, and a pipette or a straw can be used to pipette more high-purity PBMC, improving the purity and yield of PBMC.
[0028] To prevent the blood sample from being mixed with the lymphocyte separation liquid during the process of slowly flowing into the cell separation tube along the fixed ring 3 with a slope structure, the included angle between the upper surface of the fixed ring 3 and the tube wall of the separation tube body 1 is 30°, and the included angle between the lower surface of the fixed ring 3 and the tube wall of the separation tube body 1 is 75°.
[0029] In addition, to adapt to the added amount of lymphocyte separation liquid and the PBMC after centrifugation being located at the inner circle of the fixed ring 3, the volume between the lower surface of the fixed ring 3 and the bottom of the separation tube body 1 is specifically determined according to the added amount of lymphocyte separation liquid. More preferably, when the specification of the separation tube body 1 is 50 mL, the volume between the lower surface of the fixed ring 3 and the bottom of the separation tube body 1 is 15 mL.
[0030] Further, the inner diameter of the fixed ring 3 is 1 / 3 - 1 / 2 of the inner diameter of the separation tube body 1 to reduce the contact area between the separated and extracted PBMC and the lymphocyte separation liquid and the plasma layer. Furthermore, the height at the inner circle of the fixed ring 3 is 1 / 4 - 1 / 3 of the height of the separation tube body 1, and the height at the outer circle of the fixed ring 3 is 1 / 2 - 2 / 3 of the height of the separation tube body 1.
[0031] In the cell separation tube provided by the embodiment of the present application, a scale 4 is provided on the outer wall of the separation tube body 1 to facilitate the addition of a quantitatively measured volume of lymphocyte separation liquid and blood sample.
[0032] Taking the specification of the separation tube body 1 being 50 mL as an example, the usage process of the cell separation tube provided by the embodiment of the present application is as follows: Add 15 mL of lymphocyte separation liquid into the cell separation tube. At this time, the lymphocyte separation liquid is located in the gap between the lower surface of the fixed ring 3 and the bottom of the separation tube body 1. Slowly inject the blood sample to be separated into the separation tube body 1 along the fixed ring 3 with a slope structure. At this time, the blood sample is located above the lymphocyte separation liquid and does not mix with the lymphocyte separation liquid. After tightening the tube cap 2, place the cell separation tube in a centrifuge and centrifuge according to the set parameters. After centrifugation, the PBMC is in a white film shape at the inner circle of the fixed ring 3 and is located between the lymphocyte separation liquid and the plasma layer. Since the contact area between the PBMC and the lymphocyte separation liquid and the plasma layer is small, a pipette or straw can be used to quickly and accurately pipette more high-purity PBMC, reducing the mixing of other components in the blood and the lymphocyte separation liquid into the PBMC, and improving the purity and yield of the PBMC.
[0033] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, the present application can have various changes and modifications. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. A cell separation tube for separating and extracting PBMC, characterized in that, It includes a separation tube body (1) and a tube cap (2) that are detachably connected. A fixing ring (3) is provided on the inner wall of the separation tube body (1). The outer diameter of the fixing ring (3) is the same as the inner diameter of the separation tube body (1), and there is a gap between the bottom surface of the fixing ring (3) and the bottom of the separation tube body (1); both the upper and lower surfaces of the fixing ring (3) are slopes.
2. The cell separation tube for separating and extracting PBMC according to claim 1, wherein The included angle between the upper surface of the fixing ring (3) and the tube wall of the separation tube body (1) is 30°.
3. The cell separation tube for separating and extracting PBMC according to claim 1, wherein The included angle between the lower surface of the fixing ring (3) and the tube wall of the separation tube body (1) is 75°.
4. The cell separation tube for separating and extracting PBMC according to claim 1, wherein The inner diameter of the fixing ring (3) is 1 / 3 - 1 / 2 of the inner diameter of the separation tube body (1).
5. The cell separation tube for separating and extracting PBMC according to claim 1, characterized in that, The height at the inner circle of the fixing ring (3) is 1 / 4 - 1 / 3 of the height of the separation tube body (1), and the height at the outer circle of the fixing ring (3) is 1 / 2 - 2 / 3 of the height of the separation tube body (1).
6. The cell separation tube for separating and extracting PBMC according to claim 1, wherein, The volume between the lower surface of the fixing ring (3) and the bottom of the separation tube body (1) is 15 mL.
7. The cell separation tube for separating and extracting PBMC according to claim 1, characterized in that, The fixing ring (3) is a hollow fixing ring.
8. The cell separation tube for separating and extracting PBMC according to claim 1, wherein The fixing ring (3) is a solid fixing ring.
9. The cell separation tube for separating and extracting PBMC according to any one of claims 1-8, characterized in that, A scale (4) is provided on the outer wall of the separation tube body (1).