Totally-enclosed centrifugal tube

Abstract

The utility model relates to the technical field of centrifuge tubes, in particular to a fully-closed centrifuge tube which comprises a bottle body, a piston for closing the opening end of the bottle body is detachably connected to an opening in the upper end of the bottle body, and the end face of the piston communicates with a first Luer mouth, a second Luer mouth and a third Luer mouth at intervals. The lower end of the third Luer mouth is communicated with a guide pipe; and the piston can be pushed and pressed downwards to drive a sample in the bottle body to flow out from the third Luer mouth through the guide pipe. The centrifugal tube is specially designed, so that the totally-closed cell transfer process is realized, and the cell pollution risk in the transfer process is effectively reduced.

Description

Technical Field

[0001] This application relates to the technical field of centrifuge tubes, and particularly to a fully enclosed centrifuge tube. Background Art

[0002] The gene and cell therapy (CGT) industry is rapidly emerging. The gene therapy industry mainly covers several important therapies, including chimeric antigen receptor T cell therapy (CAR-T therapy), T cell receptor T cell therapy (TCR-T therapy), chimeric antigen receptor natural killer cell therapy (CAR-NK therapy), and tumor infiltrating lymphocyte therapy (TIL therapy). In these therapies, the cell products of CAR-T, TCR-T, and CAR-NK therapies must all undergo a gene editing process.

[0003] Gene editing refers to the process of modifying the genome through technical means. The electroporation platform is one of the most commonly used and effective methods for gene editing. Electroporation technology refers to the formation of transient pores in the cell membrane through the action of an electric field, allowing foreign DNA or other substances to enter the cell. In this process, the cells in the cell culture bag need to be transferred to multiple centrifuge tubes for concentration to facilitate subsequent processing and editing.

[0004] However, there is a significant problem with the conventional method: during the process of transferring cells to centrifuge tubes for concentration, the culture process cannot be fully enclosed. The cells are in a relatively enclosed sterile environment in the culture bag, but during the transfer to the centrifuge tube, the culture bag must be opened and in contact with external air, which brings potential sources of contamination and does not fully meet the production process requirements of Good Manufacturing Practice (GMP). Summary of the Utility Model

[0005] In order to reduce the risk of cell contamination during the transfer process, this application provides a fully enclosed centrifuge tube.

[0006] The fully enclosed centrifuge tube provided by this application adopts the following technical solution:

[0007] A fully enclosed centrifuge tube includes a bottle body. A piston for closing the open end of the bottle body is detachably connected to the upper open end of the bottle body. The end face of the piston is connected in series with a first Luer port, a second Luer port, and a third Luer port at intervals. A conduit is connected to the lower end of the third Luer port. The piston can be pushed downwards to drive the sample in the bottle body to flow out of the third Luer port through the conduit.

[0008] Optionally, it further includes a filter screen and a closing plug. A bacteria-filtering ventilation port is opened in the middle of the piston. The bacteria-filtering ventilation port is connected to the filter screen covering the bacteria-filtering ventilation port, and the bacteria-filtering ventilation port is detachably connected to the closing plug at the upper end of the filter screen.

[0009] Optionally, the lower end of the sealing plug is inserted into the bacteria-filtering ventilation port.

[0010] Optionally, the pore size of the filter screen is 0.20 μm - 0.25 μm.

[0011] Optionally, it further includes a push rod, which is used to press down the piston to drive the sample in the bottle body to flow out from the No. 3 Luer port through the catheter.

[0012] Optionally, the No. 1 Luer port, the No. 2 Luer port, and the No. 3 Luer port are circumferentially symmetrically arranged around the axis of the bottle body.

[0013] Optionally, the catheter is a low-adsorption catheter.

[0014] In summary, the present application includes the following beneficial technical effects:

[0015] In order to overcome the problem that the gene editing step in the traditional cell production process inevitably exposes the cells to the air, in the fully enclosed centrifuge tube provided by the present application, the traditional screw cap process is changed to a fully enclosed liquid inlet and outlet through the Luer port; in actual use, a sterile connecting tube can be used to connect the cell culture bag and the No. 1 Luer port, so as to transfer the cells in the cell culture bag into the centrifuge tube. Then, the centrifuge tube is centrifuged. After centrifugation, the supernatant is sucked out from the No. 3 Luer port and discharged into the waste liquid collection bag. An electroporation reaction solution and a substrate are added into the bottle body from the No. 2 Luer port with a syringe and mixed evenly. The No. 3 Luer port is connected to the electroporation device, and then the piston is pressed down from the outside of the bottle body with a push rod, so that the liquid flows out from the No. 3 Luer port under the action of pressure and is transferred into the electroporation device, and then electroporation can be carried out; through the special design of the centrifuge tube, a fully enclosed cell culture and gene editing process is realized in this part of the process, effectively reducing the risk of cell contamination during the transfer process, and providing safer and more reliable technical support for the development of the gene and cell therapy industry. Description of the Drawings

[0016] Figure 1 is the front view of the fully enclosed centrifuge tube in the embodiment of the present application.

[0017] Figure 2 is the top view of the fully enclosed centrifuge tube in the embodiment of the present application.

[0018] Figure 3 is the schematic diagram of the cooperation when the push rod pushes the piston in the embodiment of the present application.

[0019] Description of the reference numerals: 1, bottle body; 11, No. 1 Luer port; 12, No. 2 Luer port; 13, No. 3 Luer port; 131, catheter; 2, piston; 21, bacteria-filtering ventilation port; 22, filter screen; 23, sealing plug; 3, push rod. Detailed Embodiments

[0020] The following will further elaborate on this application in conjunction with the attached Figures 1-3 drawings for a more detailed description.

[0021] An embodiment of this application discloses a fully enclosed centrifuge tube. Referring to Figures 1 to 3 , the fully enclosed centrifuge tube includes a bottle body 1, a piston 2 inserted into the upper opening of the bottle body, and a push rod 3 for pressing the piston 2 downward from the outside of the bottle body 1. The piston 2 can completely cover the opening end of the bottle body 1.

[0022] The end face of the piston 2 is provided with spaced holes, and Luer connectors are respectively installed in the plurality of holes one by one to form a first Luer port 11, a second Luer port 12, and a third Luer port 13. The Luer connector is a standardized micro-leakage-free connector, which is connected through a male Luer connector and a matching female Luer connector part. In the embodiment of this application, taking all Luer connectors as self-sealing Luer connectors as an example, the first Luer port 11, the second Luer port 12, and the third Luer port 13 are circumferentially symmetrically arranged with the axis of the bottle body 1 as the axis, restricting the distance between the Luer ports, thereby reducing the operation difficulty.

[0023] The first Luer port 11 is connected to a cell culture bag through a sterile connecting tube. The second Luer port 12 is usually in a closed state. A conduit 131 is installed at the lower end of the third Luer port 13. The conduit 131 is a low-adsorption conduit. The upper end of the third Luer port 13 can be installed with an electroporation device connected according to actual process requirements. After pushing and pressing the piston 2 downward, the sample in the bottle body 1 can flow into the electroporation device through the conduit 131.

[0024] A bacteriostatic ventilation port 21 is opened in the middle of the piston 2, and a filter screen 22 for covering the bacteriostatic ventilation port 21 is installed at the bacteriostatic ventilation port 21. In the embodiment of this application, taking the filter screen 22 fixed at the bacteriostatic ventilation port 21 by a medical-grade biocompatible glue as an example, the filter screen 22 can be 0.20 - 0.25 μm. In the embodiment of this application, the pore size is designed to be 0.22 μm. The filter screen 22 is any conventional filter screen in the biological field that meets the processing requirements of the gene editing process and can achieve the function of "allowing gas to pass freely but blocking bacteria and other microorganisms".

[0025] A closing plug 23 for closing the bacteriostatic ventilation port 21 is detachably inserted above the filter screen 22 at the bacteriostatic ventilation port 21. Before pressing the piston 2, the closing plug 23 is not installed to keep the air circulation at the bacteriostatic ventilation port 21, and the lower end of the closing plug 23 is inserted into the bacteriostatic ventilation port 21; before pressing the piston 2, since the gas circulates at the bacteriostatic ventilation port 21, the closing plug 23 can be used for closing.

[0026] During the overall processing, the cells carry out respiration. Cellular respiration is a process by which cells obtain energy by consuming oxygen and producing carbon dioxide. For cells cultured in vitro, especially in a sealed environment, if there is insufficient oxygen or excessive accumulation of carbon dioxide in the culture environment, the growth and survival of the cells will be negatively affected. The bacteria-filtering vent 21 passes through the filter screen 22 to ensure gas exchange while preventing external pollutants from entering the cell culture system, ensuring that the cellular respiration during the processing can protect the state of the cells.

[0027] The implementation principle of a fully enclosed centrifuge tube in an embodiment of the present application is as follows: Connect the cell culture bag and the first Luer port 11 with a sterile connecting tube, thereby transferring the cells in the cell culture bag into the centrifuge tube. Do not install the closing plug 23, keep the bacteria-filtering vent 21 connected to the outside, keep the second Luer port 12 and the third Luer port 13 closed, then centrifuge the centrifuge tube. After the centrifugation is completed, connect the third Luer port 13, and suck out the supernatant from the third Luer port 13 and discharge it into the waste liquid collection bag.

[0028] Subsequently, connect the second Luer port 12, and use a syringe to add the electroporation reaction solution and the substrate into the bottle body 1 from the second Luer port 12. Use manual shaking or place the centrifuge tube in the instrument to make the centrifuge tube mix evenly by low-speed vortex. Then install the closing plug 23, and use the pushing rod 3 to press the piston 2 from the outside of the bottle body 1. Then the liquid flows out from the third Luer port 13 under the action of pressure and is transferred into the electroporation device, and electroporation can be carried out.

[0029] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited accordingly. Therefore, all equivalent changes made according to the structure, shape, and principle of the present application should be covered within the protection scope of the present application.

Claims

1. A fully enclosed centrifuge tube, characterized in that: The invention comprises a bottle body (1), wherein a piston (2) for closing the open end of the bottle body (1) is detachably connected to the upper opening of the bottle body (1), the end surface of the piston (2) is connected to a first Luer port (11), a second Luer port (12) and a third Luer port (13), and the lower end of the third Luer port (13) is connected to a conduit (131); the piston (2) can be pushed downward to drive the sample in the bottle body (1) to flow out of the third Luer port (13) through the conduit (131).

2. A fully enclosed centrifuge tube according to claim 1, characterized in that: It also includes a filter screen (22) and a sealing plug (23); a bacteria filter vent (21) is provided in the middle of the piston (2); the bacteria filter vent (21) is connected to the filter screen (22) for shielding the bacteria filter vent (21); the bacteria filter vent (21) is detachably connected to the sealing plug (23) at the upper end of the filter screen (22).

3. A fully enclosed centrifuge tube according to claim 2, characterized in that: The bacteria filtering vent (21) is inserted into the lower end of the sealing plug (23).

4. A fully enclosed centrifuge tube according to claim 2, characterized in that: The pore size of the filter (22) is 0.20 μm-0.25 μm.

5. A fully enclosed centrifuge tube according to any one of claims 1 to 4, characterized in that: It also includes a push rod (3), which is used to press down the piston (2) to drive the sample in the bottle body (1) to flow out from the No. 3 Luer port (13) through the conduit (131).

6. A fully enclosed centrifuge tube according to claim 5, characterized in that: The No. 1 Luer port (11), the No. 2 Luer port (12), and the No. 3 Luer port (13) are symmetrically arranged in the circumferential direction with the axis of the bottle body (1) as the axis.

7. A fully enclosed centrifuge tube according to claim 5, characterized in that: The conduit (131) is a low adsorption conduit.

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