A structure for extracting tumor-infiltrating lymphocytes

Abstract

This invention relates to a structure for extracting tumor-infiltrating lymphocytes (TILs), comprising a tube body, a sealing cap, a first separator, and a second separator. The tube body consists of a cell cutting section with an open top and a cell extraction section with a closed bottom, arranged sequentially from top to bottom. The cell cutting and extraction sections are rotatable and can switch transfer channels at their connection point. The first separator is fixed to the bottom of the cell cutting section and has strip-shaped first sieve holes. The second separator is detachable from the top of the cell extraction section and has strip-shaped second sieve holes. When the first and second sieve holes are aligned, the transfer channels are fully connected; when they are completely offset, the first and second separators are stacked to seal the transfer channels. This invention solves the industry problems of multiple transfers and high contamination risk in TIL extraction through a design of rotating dual sieves with dynamic on / off switching, magnetic sealing cutting, and 180° precise positioning.

Description

Technical Field

[0001] This invention relates to the field of cell extraction technology, and more specifically to a structure for extracting tumor-infiltrating lymphocytes. Background Technology

[0002] Tumor-infiltrating lymphocytes (TILs) are lymphocytes that infiltrate tumor tissue. They are a subset of lymphocytes characterized by high efficiency in recognizing tumor cell neoantigens, good homing ability, natural recognition of multiple targets, minimal side effects, and strong tumor-killing power. As a core resource for immunotherapy, the activity and purity of TILs directly determine the therapeutic effect. Traditional TIL extraction involves a multi-step process: tissue cutting → enzymatic digestion → centrifugation → cell transfer and culture. According to the "Guidelines for Quality Management of Cell Therapy Products," each additional step in the transfer operation increases the difficulty of aseptic assurance by 20%-25%. This means that each transfer increases the opportunity for cells to come into contact with the external environment, easily introducing microorganisms, impurities, and other contaminants, thus affecting the activity and quality of TIL cells, reducing extraction efficiency, and limiting the application effect of TIL cells in tumor immunotherapy.

[0003] Existing technologies employ single-chamber designs with fixed filters, enabling cutting and preliminary filtration; however, digestion and separation still require transfer to centrifuge tubes (at least two exposure risks); moreover, the filters are not adjustable, and tissue fragments easily clog them.

[0004] Therefore, how to provide a structure for extracting tumor-infiltrating lymphocytes that can reduce the number of metastases and lower the risk of contamination is a problem that urgently needs to be solved by those skilled in the art. Utility Model Content

[0005] Therefore, the purpose of this invention is to propose a structure for extracting tumor-infiltrating lymphocytes, thereby reducing the number of metastases and lowering the risk of contamination.

[0006] The technical solution of this utility model is a structure for extracting tumor-infiltrating lymphocytes, comprising:

[0007] The tube body consists of, from top to bottom, a cell cutting tube segment with an open top and a cell extraction tube segment with a closed bottom. The cell cutting tube segment and the cell extraction tube segment can rotate relative to each other, and the transfer channel can be switched at the connection point.

[0008] A sealing cap is detachable from the top opening of the cell cutting tube segment;

[0009] A first separating sieve is fixed to the bottom of the cell cutting tube segment and has strip-shaped first sieve holes on it;

[0010] The second separator is detachable from the top of the cell extraction tube segment. It has strip-shaped second sieve holes. The transfer channel is fully connected when the first sieve hole and the second sieve hole are aligned. When the first sieve hole and the second sieve hole are completely offset, the first separator and the second separator are superimposed to close the transfer channel.

[0011] According to the structure of this utility model, magnetic cutting blades are installed inside the cell cutting tube segment.

[0012] According to the structure of this utility model, the distance from the bottom end of the first separating sieve to the bottom end of the cell cutting tube is L. The cell cutting tube is sleeved within the top length range of the cell extraction tube, and the cell extraction tube has a snap-fit ​​groove. The cell cutting tube has a snap-fit ​​protrusion that snaps into the snap-fit ​​groove.

[0013] According to the structure of this utility model, the snap ring groove has two limiting deep holes, and the snap ring has two protrusions corresponding to the limiting deep holes. The two limiting deep holes and the two protrusions are initially in the same position, and the transfer channel is in a closed state. When the two protrusions are rotated 180 degrees relative to the two limiting deep holes, the transfer channel is in a fully open state.

[0014] According to the structure of this utility model, a marking strip is provided at the position of one of the limiting deep holes relative to the outer wall of the cell extraction tube segment, and positioning strips are provided at the positions of the two protrusions relative to the outer wall of the cell cutting tube segment, with the two positioning strips being 180 degrees apart in the circumferential direction.

[0015] According to the structure of this utility model, the top of the cell extraction tube section has a mounting groove for detachably connecting to the second separating screen, and the side wall of the mounting groove has a threaded structure that is adapted to the threaded structure on the screen seat of the second separating screen.

[0016] According to the structure of this utility model, the bottom end of the first separating screen or the top end of the second separating screen has a sealing groove for a sealing element.

[0017] According to the structure of this utility model, both the cell cutting tube segment and the cell extraction tube segment have an anti-adhesion coating on their inner walls.

[0018] As can be seen from the above technical solution, compared with the prior art, the beneficial effects of this utility model are as follows:

[0019] 1. This utility model achieves zero transfer operation through a rotating double-sieve structure (first separating sieve and second separating sieve): in the cutting state, the sieve holes are completely misaligned, the double sieves are stacked and sealed, and the tissue is broken up in the cutting tube section; in the transfer state, the sieve holes are aligned by rotating 180°, and the tissue falls directly into the extraction tube section; eliminating at least two transfers in the traditional process of cutting, digestion and centrifugation, reducing the risk of contamination by 60%-75%.

[0020] 2. This utility model features a 180° rotation limit: a raised point and a deep limiting hole provide a "click" positioning feedback, avoiding angular deviation; dual indication via a marking strip and a positioning strip reduces operation time. The first and second sieve holes of this utility model have a length-to-width ratio ≥3:1, increasing the passage rate of tissue fragments.

[0021] 3. The anti-adhesion coating of this utility model can be a polyethylene glycol-polylactic acid copolymer coating to reduce the cell residue rate.

[0022] 4. The sealing element of this utility model fills the gap between the double screens to prevent liquid leakage and cross-contamination. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0024] Figure 1 This invention provides a schematic diagram of a structure for extracting tumor-infiltrating lymphocytes.

[0025] Figure 2 for Figure 1 A sectional view;

[0026] Figure 3 for Figure 2 Enlarged view of part A;

[0027] Figure 4 The diagram illustrates the closed status of the transfer channel;

[0028] Figure 5 This diagram illustrates the partially connected state of the transfer channel.

[0029] Among them, 1-tube body, 11-cell cutting tube segment, 111-sealing cap, 112-first separating sieve, 1121-first sieve hole, 113-annular protrusion, 12-cell extraction tube segment, 121-second separating sieve, 1211-second sieve hole, 13-indicator strip, 14-positioning strip, 2-magnetic cutting blade, 3-sealing element. Detailed Implementation

[0030] The embodiments of this utility model are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain this utility model, and should not be construed as limiting this utility model.

[0031] In the description of this utility model, it should be understood that the terms "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0032] Traditional TIL extraction involves a multi-step process including tissue cutting, enzymatic digestion, centrifugation, and cell transfer culture. Each transfer increases the chance of cells coming into contact with the external environment, easily introducing contaminants such as microorganisms and impurities, which affects the activity and quality of TIL cells, leading to reduced extraction efficiency and limiting the application of TIL cells in tumor immunotherapy. Existing technologies employ a single-chamber structure with a fixed filter, enabling cutting and preliminary filtration; however, digestion and separation still require transfer to centrifuge tubes (at least two exposure risks); moreover, the filter is not adjustable, and tissue fragments easily clog it.

[0033] Therefore, the technical solution of this utility model is a structure for extracting tumor-infiltrating lymphocytes, see appendix. Figure 1-5 It includes: a pipe body 1, a sealing cap 111, a first partition screen 112 and a second partition screen 121.

[0034] The tube body 1 consists of a cell cutting tube segment 11 with an open top and a cell extraction tube segment 12 with a closed bottom, from top to bottom. The cell cutting tube segment 11 and the cell extraction tube segment 12 can rotate relative to each other, and the transfer channel can be switched at the connection point.

[0035] The sealing cap 111 is detachable from the top opening of the cell cutting tube segment 11;

[0036] The first separating sieve 112 is fixed to the bottom of the cell cutting tube segment 11, and has strip-shaped first sieve holes 1121 on it;

[0037] The second separating sieve 121 is detachable from the top of the cell extraction tube segment 12. It has a strip-shaped second sieve hole 1211. The first sieve hole 1121 and the second sieve hole 1211 are aligned and the transfer channel is completely connected. The first sieve hole 1121 and the second sieve hole 1211 are completely offset. The first separating sieve 112 and the second separating sieve 121 are superimposed to close the transfer channel.

[0038] This invention achieves zero-transfer operation by rotating the first and second separating screens. In the cutting state, the first and second screen holes are completely misaligned, and the double screens are superimposed and sealed, causing the tissue to be broken up within the cutting tube. In the transfer state, the screen holes are aligned by rotating 180°, and the tissue falls directly into the extraction tube. This eliminates at least two transfers in the traditional process of cutting, digestion, and centrifugation, reducing the risk of contamination by 60%-75%.

[0039] In this implementation, the specific arrangement parameters of the screen holes of the first separating screen 112 and the second separating screen 121 are as follows: the first separating screen has strip-shaped holes (length L1, width W1), and the second separating screen has strip-shaped holes (length L2 = L1, width W2 = W1), ensuring that there are no gaps when they are completely overlapped; the first screen holes and the second screen holes are evenly distributed in the middle of the separating screen, and the corresponding screen holes on the two separating screens can be offset by the distance of one screen hole. After rotation, the hole positions are precisely corresponding, and the number of holes N (N≥4 and is an even number) satisfies 180° rotational symmetry. The minimum width of the solid part on the separating screen should be ≥W1. When the two screens are misaligned, the solid part of the first screen hole must completely cover the hole area of ​​the second screen hole.

[0040] The specific parameters for the two separating screens are as follows: material: 316L stainless steel; thickness: 0.3mm; screen aperture size: 5.0mm (length) × 1.0mm (width); number of apertures: 8; minimum solid width: 14.7mm (R = 20mm); state verification: in the connected state, the two screens are aligned at 0°, and the through-hole area = 8 × 5 × 1 = 40mm². 2 Closed state: 180° rotation results in zero through-hole area (complete solid coverage); Allowable processing error: sieve hole width ±0.05mm; sieve flatness ≤0.1mm / m 2 .

[0041] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this utility model, "a plurality of" means two or more, unless otherwise explicitly specified.

[0042] See appendix Figure 2 and 3The distance from the bottom end of the first separating screen 112 to the bottom end of the cell cutting tube segment 11 is L. The cell cutting tube segment 11 is sleeved within the top length L of the cell extraction tube segment 12, and the cell extraction tube segment 12 has a snap-fit ​​groove. The cell cutting tube segment 11 has a corresponding snap-fit ​​protrusion 113 that snaps into the snap-fit ​​groove. This ensures that the transfer channel can be rotated after cutting.

[0043] While the above-mentioned solution allows rotation and changes in the position of the sieve holes, it cannot guarantee precise positioning. Therefore, in this embodiment, the locking ring groove has two limiting deep holes, and the locking protrusion ring 113 has two protrusions corresponding to the limiting deep holes. Initially, the two limiting deep holes and the two protrusions correspond to each other, and the transfer channel is in a closed state. When the two protrusions rotate 180 degrees relative to the two limiting deep holes, the transfer channel is fully open. This achieves positioning with a closed transfer channel and positioning with the transfer channel fully connected (the first and second sieve holes are all aligned).

[0044] Since the above solution is inconvenient to check whether the internal positioning is in place, this embodiment has an identification strip 14 at the position of one of the limiting deep holes relative to the outer wall of the cell extraction tube segment 12, and positioning strips 13 at the positions of the two protrusions relative to the outer wall of the cell cutting tube segment 11. The two positioning strips 13 are 180 degrees apart in the circumferential direction. Therefore, this embodiment is used to facilitate checking whether the positioning is in place.

[0045] Advantageously, the top of the cell extraction tube segment 12 has a mounting groove for detachably connecting to the second separating sieve 121. The sidewall of the mounting groove has a threaded structure that is adapted to the threaded structure on the sieve seat of the second separating sieve 121. An annular mounting chamber is formed inside the mounting groove, and either of the two sidewalls of the mounting chamber has a threaded structure.

[0046] Of course, the mounting slot and the second dividing screen 121 can also adopt a snap-fit ​​structure. The reason for adopting a threaded structure is to take into account the sealing.

[0047] More advantageously, the bottom end of the first separating screen 112 or the top end of the second separating screen 121 has a sealing groove for the sealing element 3. The sealing element 3 can be a silicone sealing ring (compression ratio 25%) to compensate for micro-gaps caused by manufacturing tolerances.

[0048] In embodiments of this invention, both the cell cutting tube segment 11 and the cell extraction tube segment 12 have an anti-adhesion coating on their inner walls. The anti-adhesion coating is a polyethylene glycol-polylactic acid copolymer coating, which reduces the cell residue rate.

[0049] A magnetic cutting blade 2 is installed inside the cell cutting tube segment 11. The magnetic cutting blade 2 is rotatably disposed inside the cell cutting tube segment 11 and located above the first partition plate to cut cell tissue samples. A magnetic drive device is magnetically driven to connect with the magnetic cutting blade 2, and the tube body can be assembled on the magnetic drive device so that the magnetic cutting blade 2 rotates to cut cell tissue samples, and its rotation axis is arranged coaxially with the axis of the cell cutting tube segment (11). The tube body can be assembled in the centrifuge device to achieve cell centrifugation separation. The bottom of the tube body has a conical structure to facilitate adaptation to the magnetic drive device and the centrifuge device.

[0050] In this utility model, unless otherwise explicitly specified and limited, the terms "installation," "connection," "joining," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0051] Figure 4 The diagram illustrates the closed status of the transfer channel; Figure 5 The diagram illustrates the partially connected state of the transfer channel; the term "partially" means that the corresponding screen holes on the first and second separating screens are between 0 and 180 degrees, neither completely closed nor completely open.

[0052] In this invention, the cell cutting tube segment 11 and the cell extraction tube segment 12 are connected to the snap-fit ​​groove via a snap ring 113. Figure 3 Initial state: The first sieve hole 1121 and the second sieve hole 1211 are completely misaligned, and the two sieves are superimposed to form a closed bottom surface. Figure 4 During the channel switching operation, hold the cell cutting tube segment 11 and rotate it 180° until the positioning strip 13 is aligned with the marker strip 14; at this time, the protrusion is engaged with another limiting deep hole, the sieve holes are fully aligned, and the tissue fragments fall into the extraction tube segment 12 through the channel.

[0053] The magnetic cutting system uses a magnetic cutting blade made of medical-grade 316L stainless steel with an edge thickness of ≤0.1mm. Operating parameters: the magnetic drive unit operates at a speed of 300rpm ±50rpm, and the cutting time is 10-15 minutes.

[0054] Rotary positioning mechanism with a limiting deep hole diameter of 1.2mm and a depth of 2mm; a protrusion diameter of 1.0mm (interference fit provides tactile feedback); a "click" sound indicates that it has reached the correct position when rotated.

[0055] The operation process of this utility model is as follows:

[0056] In the S1 tissue cutting stage, open the sealing cap 111, put in the tumor tissue (≤1g) and add an appropriate amount of PBS solution containing double antibodies, and close the sealing cap 111; start the magnetic stirrer, and rotate the magnetic cutting blade at 300rpm for 15 minutes to cut the tumor tissue into a paste.

[0057] S2 Tissue transfer and cleaning: Rotate the cutting tube segment 180° until the sieve holes are aligned, allowing the tissue to leak into the extraction tube segment 12. At the same time, add an appropriate amount of PBS to rinse the first separating sieve, so that all tissues leak into the extraction tube segment 12. Remove the magnetic cutter and close the sealing cap.

[0058] S3 centrifugation, digestion and culture: rotate the tube segment to a closed state, place it in a centrifuge (1500g, 10 minutes), wash the tumor tissue, discard the supernatant; add digestive enzymes, close the sealing cap, place it in a carbon dioxide incubator for digestion, after digestion is completed, centrifuge to obtain cells, and sort and culture them.

[0059] Throughout the entire operation, the device is operated in a sterile environment. This multi-step process reduces the number of tissue and cell transfers, thereby minimizing the risk of contamination. Total operation time: ≤60 minutes (traditional methods ≥120 minutes), cell viability can be greater than ≥92%.

[0060] This invention achieves breakthroughs in pollution control, operational efficiency, cell activity, and ease of maintenance through innovative designs such as rotating double screen dynamic on / off, magnetic sealing cutting, and 180° precise positioning, solving the industry problems of multiple transfers and high pollution risk in TIL extraction.

[0061] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can combine and integrate the different embodiments or examples described in this specification.

[0062] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A structure for extracting tumor infiltrating lymphocytes, characterized by, include: The tube body (1) consists of a cell cutting tube segment (11) with an open top and a cell extraction tube segment (12) with a closed bottom, from top to bottom. The cell cutting tube segment (11) and the cell extraction tube segment (12) can rotate relative to each other and can switch transfer channels at the connection point. A sealing cap (111) is detachable from the top opening of the cell cutting tube segment (11); A first separating sieve (112) is fixed to the bottom of the cell cutting tube segment (11) and has a strip-shaped first sieve hole (1121) thereon; The second separating sieve (121) is detachable from the top of the cell extraction tube segment (12) and has a strip-shaped second sieve hole (1211). The first sieve hole (1121) and the second sieve hole (1211) are aligned and the transfer channel is fully connected. The first sieve hole (1121) and the second sieve hole (1211) are completely offset. The first separating sieve (112) and the second separating sieve (121) are superimposed to close the transfer channel.

2. The structure for extracting tumor infiltrating lymphocytes according to claim 1, wherein, The cell cutting tube segment (11) is equipped with a magnetic cutting blade (2).

3. The structure for extracting tumor infiltrating lymphocytes according to claim 1, wherein, The distance from the bottom end of the first separating screen (112) to the bottom end of the cell cutting tube segment (11) is L. The cell cutting tube segment (11) is sleeved on the top of the cell extraction tube segment (12) within the length range of L. The cell extraction tube segment (12) has a snap-fit ​​groove, and the cell cutting tube segment (11) has a snap-fit ​​protrusion (113) that snaps into the snap-fit ​​groove.

4. The structure for extracting tumor infiltrating lymphocytes according to claim 3, wherein The snap ring groove has two limiting deep holes, and the snap ring (113) has two protrusions corresponding to the limiting deep holes. The two limiting deep holes and the two protrusions are initially in the same position, and the transfer channel is in a closed state. When the two protrusions are rotated 180 degrees relative to the two limiting deep holes, the transfer channel is in a fully open state.

5. The structure for extracting tumor infiltrating lymphocytes according to claim 4, wherein A marker strip (14) is provided at the position of one of the limiting deep holes relative to the outer wall of the cell extraction tube segment (12), and a positioning strip (13) is provided at the positions of the two protrusions relative to the outer wall of the cell cutting tube segment (11). The two positioning strips (13) are 180 degrees apart in the circumferential direction.

6. The structure for extracting tumor-infiltrating lymphocytes according to claim 3, characterized in that, The top of the cell extraction tube segment (12) has a mounting groove for detachably connecting to the second separating screen (121). The side wall of the mounting groove has a threaded structure that is adapted to the threaded structure on the screen seat of the second separating screen (121).

7. The structure for extracting tumor infiltrating lymphocytes according to claim 6, wherein The bottom end of the first separating screen (112) or the top end of the second separating screen (121) has a sealing groove of the sealing element (3).

8. The structure for extracting tumor infiltrating lymphocytes according to any one of claims 1 to 7, wherein Both the cell cutting tube segment (11) and the cell extraction tube segment (12) have an anti-adhesion coating on their inner walls.

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