CN215328073U - Integrated vacuum collection tube for separating peripheral blood mononuclear cells - Google Patents

Abstract

The utility model belongs to the technical field of medical instruments, and particularly relates to an integrated vacuum collection tube for separating Peripheral Blood Mononuclear Cells (PBMC). Used for separating, storing and collecting peripheral blood mononuclear cells and pre-separation thereof; comprises a cap, a tube and an isolation component; the cap is detachably connected with the upper pipe body; the pipe body comprises an upper pipe body and a lower pipe body; the isolation component is arranged between the upper tube body and the lower tube body, the upper tube body is used for pre-placing the anticoagulant, and the lower tube body is used for pre-placing the lymphocyte separation fluid. After blood collection is finished according to blood collection specifications, a Xin whole blood sample mixed with an anticoagulant can be obtained, the blood collection tube is directly centrifuged at high speed, and PBMC is separated above the isolation part to form a leucoderma layer and is close to the isolation part; the lower tube body is filled with impurities such as lymphocyte separation liquid, red blood cells and the like. The collection tube is designed by matching an isolation part with a solution, a PBMC separation process is realized in one step, and the extraction time of PBMC is shortened by about half.

Description

Integrated vacuum collection tube for separating peripheral blood mononuclear cells

Technical Field

The utility model belongs to the technical field of medical instruments, and particularly relates to an integrated vacuum collection tube for separating peripheral blood mononuclear cells.

Background

Peripheral Blood Mononuclear Cells (PBMC) are cells with a single nucleus in Peripheral blood, including lymphocytes and monocytes. Note here that monruclear cell monocytes, rather than Monocyte monocytes. The main separation method of peripheral blood mononuclear cells is a Ficoll-hypaque (polysucrose-diatrizoate) density gradient centrifugation method, and the operation steps are as follows:

1. peripheral venous blood is taken, anticoagulated by heparin, added with equal amount of Hank's solution and mixed evenly.

2. The layering solution was added to the tube in an amount of about 1/2% of the blood volume. The blood was slowly added to the stratified liquid surface along the vessel wall with a pipette.

3. Placing in a horizontal centrifuge for centrifugation at 1500r/min for 10 min. The vast majority of mononuclear cells are seen suspended at the interface between plasma and stratified fluid, in the form of white membranes.

4. Sucking the cell layer of the interface with a tip pipette, placing in another test tube, adding appropriate amount of Hank's solution, mixing, centrifuging at 1000r/min for 10min, and discarding the supernatant.

5. Cells were washed 2 times in the same manner.

6. The cells were resuspended in RPMI-1640 medium and stored at 4 ℃ until use.

Therefore, the conventional PBMC extraction process is complex and complicated, the optimized steps also need more than ten steps, the time is 1.5-2 hours, and especially when a plurality of samples exist, the time window required for extraction cannot be met.

SUMMERY OF THE UTILITY MODEL

In view of the above, the present invention provides an integrated vacuum collection tube for separating peripheral blood mononuclear cells, which is designed by an isolation component, and realizes a PBMC separation process in one step, and the extraction time of PBMCs is shortened by about half.

In order to achieve the technical purpose, the utility model adopts the following specific technical scheme:

an integrated vacuum collection tube for separating peripheral blood mononuclear cells is used for collecting, storing and separating the peripheral blood mononuclear cells and pre-separation substances thereof; comprises a cap, a tube and an isolation component; the cap is detachably connected with the upper pipe body; the pipe body comprises an upper pipe body and a lower pipe body; the isolation component is arranged between the upper pipe body and the lower pipe body, and one end, close to the upper pipe body, of the isolation component is provided with an anticoagulant placing position; after the peripheral blood mononuclear cells are separated, the anticoagulant on the anticoagulant placing position and the isolation component enable the separated peripheral blood mononuclear cells and serum to be located in the upper tube body, the peripheral blood mononuclear cells are close to the isolation component, and the lymphocytes are located in the lower tube body; the lower tube body is filled with lymphocyte separation liquid.

Further, the isolation member is in a conical surface shape.

Further, the allowable aperture of the isolation component is smaller than the diameter of peripheral blood mononuclear cells, and the allowable aperture is larger than lymphocytes.

Further, the detachable connection is a threaded connection or a snap connection.

Further, the upper pipe body and the lower pipe body are of an integrated structure.

Furthermore, one end of the cap, which is far away from the upper tube body, is provided with a collection port; the evacuated collection tube further comprises a plug for removably sealing the collection port.

Furthermore, a puncture needle through hole is also arranged on the tube plug; the puncture needle through hole is used for enabling the puncture needle to pass through and is closed when the puncture needle leaves the tube plug.

Further, the tube plug is a rubber plug.

Further, the size of a proximal end face of the rubber plug in the direction from the upper pipe body to the lower pipe body is smaller than that of a distal end face.

Further, a pre-isolate of the peripheral blood mononuclear cells is located over the insulating member.

By adopting the technical scheme, the utility model can obtain the following beneficial effects:

1. adding peripheral blood mononuclear cells and pre-separation thereof above the isolation component, and uniformly mixing the peripheral blood mononuclear cells and the pre-separation thereof with an anticoagulant; then, after centrifugation by a centrifuge, peripheral blood mononuclear cells are concentrated above the isolation component, and impurities such as lymphocytes and erythrocytes are concentrated below the isolation component; at the moment, the separated peripheral blood mononuclear cells can be obtained by penetrating the blood serum through the puncture needle, and then can enter subsequent application after being frozen, so that compared with the traditional steps, the process is greatly simplified, and the separation time is saved.

2. According to the utility model, no additional suction tube or pipetting device is needed for transferring when PBMC is separated, so that cross contamination caused by pipetting is avoided; (b) sample confusion caused by transfer errors; (c) the medical staff exposed in the process of pipetting can be directly centrifuged at high speed.

3. After the centrifugation of the collection tube, the separated white membrane layer can be directly sucked by a gun head or a disposable suction tube, namely PBMC cells can be obtained, and the PBMC cells can be used for subsequent cleaning and cryopreservation.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

FIG. 1 is a schematic view of an integrated vacuum collection tube for separating peripheral blood mononuclear cells according to an embodiment of the present invention;

FIG. 2 is a schematic diagram illustrating the effect of an integrated vacuum collection tube for separating peripheral blood mononuclear cells before centrifugation of a peripheral blood mononuclear cell pre-separator according to an embodiment of the present invention;

FIG. 3 is a schematic diagram illustrating the effect of an integrated vacuum collection tube for separating peripheral blood mononuclear cells after centrifugation of a peripheral blood mononuclear cell pre-separation according to an embodiment of the present invention;

wherein: 1. capping; 11. a pipe plug; 2. a pipe body; 3. an insulating member; 4. an anticoagulant; 5. lymphocyte separation liquid; 6. and (5) a white film layer.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. It is to be understood that the described embodiments are merely exemplary of the utility model, and not restrictive of the full scope of the utility model. The utility model is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It is noted that various aspects of the embodiments are described below within the scope of the appended claims. It should be apparent that the aspects described herein may be embodied in a wide variety of forms and that any specific structure and/or function described herein is merely illustrative. Based on the disclosure, one skilled in the art should appreciate that one aspect described herein may be implemented independently of any other aspects and that two or more of these aspects may be combined in various ways. For example, an apparatus may be implemented and/or a method practiced using any number of the aspects set forth herein. Additionally, such an apparatus may be implemented and/or such a method may be practiced using other structure and/or functionality in addition to one or more of the aspects set forth herein.

It should be noted that the drawings provided in the following embodiments are only for illustrating the basic idea of the present invention, and the drawings only show the components related to the present invention rather than the number, shape and size of the components in practical implementation, and the type, quantity and proportion of the components in practical implementation can be changed freely, and the layout of the components can be more complicated.

In addition, in the following description, specific details are provided to facilitate a thorough understanding of the examples. However, it will be understood by those skilled in the art that the aspects may be practiced without these specific details.

In one embodiment of the utility model, an integrated vacuum collection tube for separating peripheral blood mononuclear cells is provided, which is used for collecting, storing and separating the peripheral blood mononuclear cells and pre-separation thereof; as shown in fig. 1, comprises a cap 1, a tube 2 and an insulating member 3; the cap 1 is detachably connected with the upper pipe body 2; the pipe body 2 comprises an upper pipe body 2 and a lower pipe body 2; the isolation component 3 is arranged between the upper pipe body 2 and the lower pipe body 2, and one end close to the upper pipe body 2 is provided with a anticoagulant 4 placing position; after the separation of the peripheral blood mononuclear cells, the anticoagulant 4 and the isolation component 3 on the position where the anticoagulant 4 is placed enable the separated peripheral blood mononuclear cells and serum to be located in the upper tube body 2, the peripheral blood mononuclear cells are close to the isolation component 3, and the lymphocytes are located in the lower tube body 2; the lower tube body 2 is filled with lymphocyte separation liquid 5.

In the present embodiment, the insulating member 3 has a conical surface shape.

In this embodiment, the isolation member 3 has a smaller diameter than the mononuclear cell diameter of peripheral blood, and a larger diameter than the lymphocytes, and in this embodiment, a conventional lymphocyte filter is used.

In this embodiment, the detachable connection is a threaded connection or a snap connection.

In this embodiment, the upper tube 2 and the lower tube 2 are of an integral structure.

In this embodiment, a collection port is disposed at one end of the cap 1 away from the upper tube body 2; the evacuated collection tube further comprises a stopcock 11, the stopcock 11 being for removably sealing the collection port.

In this embodiment, the plug 11 is further provided with a puncture needle passing hole; the puncture needle passing hole is used for passing the puncture needle, and the puncture needle is closed when the puncture needle leaves the tube plug 11.

In this embodiment, the plug 11 is a rubber plug.

In this embodiment, the rubber plug has a proximal end face size smaller than a distal end face size in a direction from the upper pipe body 2 to the lower pipe body 2.

In this embodiment, the pre-separation of peripheral blood mononuclear cells is located above the barrier member 3.

The pipe cap and the pipe plug 11 of the present embodiment are all made by the prior art. The isolation component 3 is conical and fixed inside the tube body 2, and the connection strength can ensure that lymphocytes are filtered out during centrifugation

Preparing a result:

the lymphocyte separation liquid 5 is filled at the bottom of the tube body 2; an anticoagulant 4 is arranged above the isolation component 3; pre-separated cells of PBMC are filled above the insulating member 3; screwing on the pipe cap; plugging a pipe plug 11; (as shown in FIG. 2)

And (3) a separation process:

1. collecting blood according to specification, adding into upper tube, and mixing with anticoagulant

2. Placing the vacuum collection tube into a centrifuge for centrifugation until a white film layer 6 is generated above the isolation component 3 (as shown in figure 3);

3. and taking the vacuum collecting tube out of the centrifuge, sucking the leucocyte layer 6 by using a pipette, collecting the PBMC in the leucocyte layer 6, and then washing and freezing the PBMC for subsequent use.

The above description is only for the specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (10)

1. An integral type vacuum collection pipe of separation peripheral blood mononuclear cell characterized in that: for collecting, storing and separating PBMC and pre-isolates thereof; comprises a cap, a tube and an isolation component; the cap is detachably connected with the upper pipe body; the pipe body comprises an upper pipe body and a lower pipe body; the isolation component is arranged between the upper pipe body and the lower pipe body, and one end, close to the upper pipe body, of the isolation component is provided with an anticoagulant placing position; after the peripheral blood mononuclear cells are separated, the anticoagulant on the anticoagulant placing position and the isolation component enable the separated peripheral blood mononuclear cells and serum to be located in the upper tube body, and the peripheral blood mononuclear cells are close to the isolation component; the lower tube body is filled with lymphocyte separation liquid.

2. The unitary evacuated collection tube of claim 1, wherein: the isolation component is in a conical surface shape.

3. The unitary evacuated collection tube of claim 1, wherein: the allowable aperture of the isolation component is smaller than the diameter of peripheral blood mononuclear cells, and the allowable aperture is larger than lymphocytes.

4. The unitary evacuated collection tube of claim 1, wherein: the detachable connection is a threaded connection or a snap connection.

5. The unitary evacuated collection tube of claim 1, wherein: the upper pipe body and the lower pipe body are of an integrated structure.

6. The unitary evacuated collection tube of claim 1, wherein: one end of the cap, which is far away from the upper pipe body, is provided with a collection port; the evacuated collection tube further comprises a plug for removably sealing the collection port.

7. The unitary evacuated collection tube of claim 6, wherein: the plug is also provided with a puncture needle through hole; the puncture needle through hole is used for enabling the puncture needle to pass through and is closed when the puncture needle leaves the tube plug.

8. The unitary evacuated collection tube of claim 7, wherein: the pipe plug is a rubber plug.

9. The unitary evacuated collection tube of claim 8, wherein: the size of a proximal end face of the rubber plug in the direction from the upper pipe body to the lower pipe body is smaller than that of a distal end face.

10. The unitary evacuated collection tube of claim 1, wherein: the pre-isolate of peripheral blood mononuclear cells is located over the insulating member.

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