A device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis
By designing an L-shaped base and a filtration and separation component, and utilizing the deformable characteristics of the flexible filter, the problem of separation difficulties caused by PBS buffer leakage was solved, achieving efficient separation and collection of epithelial and stromal cells and simplifying laboratory operations.
Patent Information
- Application Number
- CN202522117023.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-30
- Publication Date
- 2026-08-25
- Estimated Expiration
- 2035-09-30
AI Technical Summary
When separating ectopic epithelial and stromal cells from human adenomyosis using existing cell filter structures, PBS buffer tends to flow along the outer wall of the centrifuge tube onto the worktable, causing inconvenience to the separation process and requiring additional cleaning.
A device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis was designed. It adopts an L-shaped base, a fixing tube, a centrifuge tube, and a filtration and separation component, including a mounting ring, a positioning ring, a flexible filter, and a ring frame. By utilizing the deformable characteristics of the flexible filter, the separation and collection of cells can be achieved by adjusting the shape of the filter, reducing cleaning work.
It simplifies the cell separation process, avoids PBS buffer from flowing onto the worktable, improves separation efficiency and flexibility of use, and is suitable for laboratory culture operations.
Smart Images
Figure CN224678065U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of medical device technology, and in particular to a device for isolating and culturing ectopic epithelial and stromal cells in human adenomyosis. Background Technology
[0002] Adenomyosis, or endometrial glands and stroma, is a disease caused by the invasion of endometrial glands and stroma into the myometrium, and is a special type of endometriosis. Because primary cultured cells closely resemble and reflect in vivo growth characteristics, they are now widely used in various experimental studies. Therefore, isolating and culturing ectopic epithelial and stromal cells from adenomyosis lesions is particularly important.
[0003] In the prior art, when separating epithelial cells and mesenchymal cells, a cell filter is placed at the top of a centrifuge tube, the cell mixture is placed inside the filter, and the cells are rinsed with PBS buffer. The cells that remain inside the filter are epithelial cells, while the cells that pass through the filter into the centrifuge tube are mesenchymal cells. Then, the filter is flipped over onto another centrifuge tube, rinsed with PBS buffer, and the epithelial cells are collected into the centrifuge tube. After separation, the cells are placed in a centrifuge for processing, the supernatant is discarded, and then the cells are cultured in a culture medium.
[0004] However, the existing cell filter structure is relatively simple and the filter shape is fixed. When rinsing by flipping the cap, PBS buffer can easily flow along the outer wall of the centrifuge tube onto the operating table, affecting the separation operation. It also needs to be cleaned afterward, making it inconvenient to use.
[0005] Therefore, there is an urgent need to develop a novel device suitable for the isolation and culture of ectopic epithelial and stromal cells in human adenomyosis. Utility Model Content
[0006] The purpose of this invention is to overcome the defects of the existing technology and provide a device for isolating and culturing ectopic epithelial and stromal cells in human adenomyosis.
[0007] The objective of this utility model can be achieved through the following technical solutions:
[0008] This utility model provides a device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis, including an L-shaped base, a pair of fixed tubes disposed on the bottom surface of the L-shaped base, centrifuge tubes disposed in the fixed tubes respectively, and a filter separation component disposed at the top of the centrifuge tubes.
[0009] The two centrifuge tubes are used to collect mesenchymal cells and epithelial cells, respectively, and each centrifuge tube is connected to a cap at the top.
[0010] The filtration and separation assembly includes an installation ring, a positioning ring, a flexible filter screen, an annular frame, and a positioning mechanism; the installation ring is horizontally positioned, and the positioning ring is positioned around the periphery of the installation ring and is adapted to the top of the centrifuge tube;
[0011] The filter screen is located at the bottom of the mounting ring, the bottom of the ring frame is connected to the bottom surface of the flexible filter screen, and there is a gap between the top of the ring frame and the mounting ring, and the ring frame is limited by the positioning mechanism.
[0012] Furthermore, a pair of limiting plates are provided on the vertical surface of the L-shaped base near the point of rotation.
[0013] Furthermore, the bottom of the limiting plate is provided with an arc-shaped limiting groove, the curvature of which matches the outer edge of the top cover.
[0014] Furthermore, the L-shaped base has a locking block on its vertical surface directly above the limiting plate, which is used to limit the position of the limiting plate.
[0015] Furthermore, a handle is horizontally connected to the outer wall of the mounting ring.
[0016] Furthermore, the inner diameter of the positioning ring is adapted to the outer diameter of the centrifuge tube opening.
[0017] Furthermore, a clearance opening is provided on one side of the positioning ring to allow space for the top cover.
[0018] Furthermore, the flexible filter is a deformable filter that can deform in the opposite direction under stress.
[0019] Furthermore, the pore size of the flexible filter is 30-50 μm.
[0020] Furthermore, the sidewall of the ring frame is provided with a notch for the flow of cell fluid.
[0021] Furthermore, the positioning mechanism includes a set screw threaded through the outer wall of the mounting ring, a protrusion fixed to the inner wall of the mounting ring, and a circular groove and a recess respectively opened on the outer walls of both sides of the ring frame.
[0022] Furthermore, one end of the set screw can abut against the circular groove, and the protrusion can abut against the recess.
[0023] Furthermore, parallel guide blocks are symmetrically arranged on the vertical surface of the L-shaped base.
[0024] Furthermore, the parallel guide blocks are slidably connected by a shield.
[0025] Compared with the prior art, the present invention has the following beneficial effects:
[0026] (1) This utility model innovatively combines centrifuge tubes and filter separation components, and takes advantage of the deformable characteristics of the flexible filter screen. The cell mixture can be placed into the filter screen to rinse and separate interstitial cells. After adjusting the shape of the filter screen, it can be rinsed in the opposite direction to collect epithelial cells. This simplifies the separation process of ectopic epithelial and interstitial cells in human adenomyosis, reduces cleaning work, and is suitable for laboratory culture operations.
[0027] (2) This utility model cleverly uses the structure of ring frame, filter and top wire to adjust the shape of the filter, avoids the problem of PBS buffer flowing onto the operating table, and makes the separation of epithelial cells and mesenchymal cells more convenient.
[0028] (3) The human adenomyosis ectopic epithelial and stromal cell separation and culture device in this utility model can be adjusted according to the style of the centrifuge tube. If the centrifuge tube is a split type, that is, the centrifuge tube body and the top cover are threaded together, then there is no need to use a limiting plate. If the centrifuge tube is a snap-on type, the limiting plate can be used to limit the top cover to avoid affecting the operator's separation operation. It is flexible and convenient to use. Attached Figure Description
[0029] Figure 1 This is a schematic diagram of the overall three-dimensional structure of this utility model.
[0030] Figure 2 This is a schematic diagram of the structure of this utility model without centrifuge tubes.
[0031] Figure 3 This is a schematic diagram of the structure of the filtration and separation component of this utility model.
[0032] Figure 4 This is a partial cross-sectional view of the filter separation component of this utility model.
[0033] Figure 5 This is a schematic diagram of the filter screen after it has been reversed according to the present invention.
[0034] Explanation of markings in the diagram:
[0035] 1-L-shaped base; 2-Fixing tube; 3-Centrifuge tube; 4-Top cover; 5-Guide block; 6-Shield; 7-Limiting plate; 8-Limiting groove; 9-Clocking block; 10-Mounting ring; 11-Positioning ring; 12-Leaving opening; 13-Flexible filter screen; 14-Ring frame; 15-Handle; 16-Top screw; 17-Round groove; 18-Protrusion; 19-Groove. Detailed Implementation
[0036] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. This embodiment is based on the technical solution of the present invention and provides detailed implementation methods and specific operating procedures; however, the scope of protection of the present invention is not limited to the following embodiments.
[0037] In this utility model, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicating orientation or positional relationships, are based on the orientation or positional relationships shown in the accompanying drawings and 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, and therefore should not be construed as a limitation of this utility model; the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance; furthermore, unless otherwise explicitly specified and limited, the terms "installed," "connected," and "linked" should be interpreted broadly, for example, they can be fixed connections, detachable connections, or integral connections; they can be mechanical connections or electrical connections; they can be direct connections or indirect connections through an intermediate medium; they can be internal connections between two components. For those skilled in the art, the specific meaning of the above terms in this utility model can be understood according to the specific circumstances.
[0038] It should be noted that similar labels and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.
[0039] Example 1:
[0040] This embodiment provides a device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis. The device includes an L-shaped base 1, a pair of fixed tubes 2 disposed on the bottom surface of the L-shaped base 1, centrifuge tubes 3 disposed in the fixed tubes 2, and a filter separation component disposed at the top of the centrifuge tubes 3.
[0041] In this embodiment, the two centrifuge tubes 3 are used to collect interstitial cells and epithelial cells, respectively, and the top of each centrifuge tube 3 is connected to a top cap 4 for sealing the opening of the centrifuge tube after collection.
[0042] The filtration and separation assembly of this embodiment includes a mounting ring 10, a positioning ring 11, a flexible filter screen 13, an annular frame 14, and a positioning mechanism. The mounting ring 10 is horizontally positioned, and the positioning ring 11 is arranged around the periphery of the mounting ring 10 and is adapted to the top end of the centrifuge tube 3. The flexible filter screen 13 is located at the bottom of the mounting ring 10, and the bottom of the annular frame 14 is connected to the bottom surface of the flexible filter screen 13. A gap is provided between the top of the annular frame 14 and the mounting ring 10, and the annular frame 14 is limited by the positioning mechanism.
[0043] Example 2:
[0044] This embodiment provides a device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis. The device includes an L-shaped base 1, a pair of fixed tubes 2 disposed on the bottom surface of the L-shaped base 1, centrifuge tubes 3 disposed in the fixed tubes 2, and a filter separation component disposed at the top of the centrifuge tubes 3.
[0045] The difference from Embodiment 1 is that, in this embodiment, a pair of limiting plates 7 are provided on the vertical surface of the L-shaped base 1 near the rotation. The bottom of the limiting plate 7 is provided with an arc-shaped limiting groove 8, and the curvature of the arc-shaped limiting groove 8 matches the outer edge of the top cover 4. On the vertical surface of the L-shaped base 1, a locking block 9 is provided directly above the limiting plate 7 for limiting the position of the limiting plate 7.
[0046] In this embodiment, a handle 15 is horizontally connected to the outer wall of the mounting ring 10. The inner diameter of the positioning ring 11 is adapted to the outer diameter of the centrifuge tube 3, and a clearance opening 12 is provided on one side of the positioning ring 11 to provide clearance for the top cover 4. The flexible filter 13 is a deformable filter that can deform in the opposite direction under force, and the pore size of the flexible filter 13 is 30-50 μm, preferably 40 μm. The flexible filter 13 is commercially available. A notch is provided on the side wall of the ring frame 14 to allow cell fluid to flow.
[0047] The positioning mechanism of this embodiment includes a set screw 16 threaded through the outer wall of the mounting ring 10, a protrusion 18 fixed to the inner wall of the mounting ring 10, and a circular groove 17 and a recess 19 respectively formed on the outer walls of both sides of the annular frame 14. One end of the set screw 16 can abut against the circular groove 17, and the protrusion 18 can abut against the recess 19. When one end of the set screw 16 abuts against the circular groove 17, the protrusion 18 can be pressed against the recess 19, thereby achieving effective limiting of the annular frame 14 by the positioning mechanism. When the set screw 16 is loosened, the limiting is engaged.
[0048] Example 3:
[0049] This embodiment provides a device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis. The device includes an L-shaped base 1, a pair of fixed tubes 2 disposed on the bottom surface of the L-shaped base 1, centrifuge tubes 3 disposed in the fixed tubes 2, and a filter separation component disposed at the top of the centrifuge tubes 3.
[0050] The difference from Embodiment 1 is that, in this embodiment, parallel guide blocks 5 are symmetrically arranged on the vertical surface of the L-shaped base 1, and a cover 6 is slidably connected between the parallel guide blocks 5. The cover 6 can move up and down along the guide blocks 5 to cover the operating area and prevent dust when idle.
[0051] Example 4:
[0052] This embodiment provides a device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis. The device includes an L-shaped base 1, a pair of fixed tubes 2 disposed on the bottom surface of the L-shaped base 1, centrifuge tubes 3 disposed in the fixed tubes 2, and a filter separation component disposed at the top of the centrifuge tubes 3.
[0053] Reference Figures 1-5 In this embodiment, two cylindrical fixing tubes 2 are vertically fixed to the bottom surface of the horizontal section of the L-shaped base 1. Centrifuge tubes 3 are placed inside the fixing tubes 2, and a detachable top cover 4 is fitted to the top of the centrifuge tubes 3. Two limiting plates 7 are rotatably connected to the vertical section of the L-shaped base 1 near the fixing tubes 2, and the limiting plates 7 are rotated via a pivot. An arc-shaped limiting groove 8 is formed at the bottom of the limiting plate 7, and the curvature of the limiting groove 8 matches the outer edge of the top cover 4 of the centrifuge tube 3. Two pairs of locking blocks 9 are also fixed on the same side of the L-shaped base 1, and the locking blocks 9 are symmetrically distributed on both sides of the limiting plate 7 for fixing the limiting plate 7.
[0054] In the filtration and separation assembly of this embodiment, a mounting ring 10 is provided at the top of the centrifuge tube 3, and a coaxial positioning ring 11 is integrally formed on its surface and bottom. The inner diameter of the positioning ring 11 is adapted to the outer diameter of the centrifuge tube 3 opening, and a rectangular clearance opening 12 is provided on one side of the positioning ring 11 to provide clearance for the top cover 4.
[0055] A flexible filter 13 is provided at the bottom of the mounting ring 10. The edge of the flexible filter 13 is connected to the mounting ring 10. The filter pore size is 40μm, which can be used to separate epithelial cells and stromal cells. A ring frame 14 is coaxially arranged inside the flexible filter 13. The ring frame 14 is a rigid polymer ring, and its bottom is bonded and fixed to the inner wall of the bottom of the flexible filter 13. A gap is provided between the ring frame 14 and the inner wall of the mounting ring 10. A handle 15 for easy gripping is provided horizontally on the outer wall of the mounting ring 10, and the surface is provided with anti-slip texture.
[0056] The positioning mechanism of this embodiment includes a set screw 16, which is threaded through the side wall of the mounting ring 10 and has a tapered end; a circular groove 17, which is formed on the outer wall of the ring frame 14; a protrusion 18, which is vertically fixed to the inner wall of the mounting ring 10; and a groove 19, which is formed on the outer wall of the ring frame 14.
[0057] In the positioned state, rotating the set screw 16 causes its end to embed into the circular groove 17, thereby driving the protrusion 18 to engage with the recess 19. At this time, the annular frame 14 and the mounting ring 10 form a rigid connection, and the flexible filter screen 13 maintains its unfolded shape.
[0058] In the released state, the top screw 16 is rotated in the opposite direction to exit the circular groove 17, and the protrusion 18 no longer has a limiting effect on the annular frame 14. Then, the annular frame 14 is pulled upward, and the flexible filter screen 13 can shrink and deform under the tension due to its deformability, and unfold in the opposite direction.
[0059] In addition, two parallel guide blocks 5 are fixedly provided on one side of the vertical section of the L-shaped base 1 in this embodiment. A transparent cover 6 is slidably connected between the two guide blocks 5. The cover 6 can move up and down along the guide blocks 5 to cover the operating area and prevent dust when idle.
[0060] The operating procedure for the human adenomyosis ectopic epithelial and stromal cell isolation and culture device in this embodiment is as follows:
[0061] When separating ectopic epithelial and stromal cells from human adenomyosis on a sterile operating table, the mounting ring 10 is fastened to the top of the centrifuge tube 3 via the positioning ring 11, and the flexible filter 13 is suspended inside the centrifuge tube 3. The positioning mechanism is in the locked state. The tissue mixture is injected into the flexible filter 13, and PBS buffer is injected into the flexible filter 13 with a syringe to rinse it. The stromal cells pass through the flexible filter 13 with the PBS buffer and enter the centrifuge tube 3, while the epithelial cells remain on the surface of the flexible filter 13.
[0062] After the liquid has dripped completely, hold handle 15 to lift the mounting ring 10 and move it above another centrifuge tube 3. Rotate the set screw 16 to disengage it from the circular groove 17, releasing the limiting position of the positioning mechanism, and pull the annular frame 14 upward. The flexible filter screen 13 is pulled and contracted by the annular frame 14, passing through the inner hole of the mounting ring 10 and flipping outward, with the flipped shape as shown. Figure 5 As shown ( Figure 5 (A schematic diagram of the ring frame 14 after it has been pulled out and rotated 180°).
[0063] The inverted flexible filter 13 and the ring frame 14 are assembled together with another centrifuge tube 3. PBS buffer is injected in reverse to rinse the tube, allowing the epithelial cells to detach from the flexible filter 13 and enter the centrifuge tube 3. The two centrifuge tubes 3 are then sealed and centrifuged. The supernatant is removed, and the tubes are finally placed in culture medium for further culture.
[0064] The above description of the embodiments is provided to enable those skilled in the art to understand and use the utility model. It will be apparent to those skilled in the art that various modifications can be easily made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present utility model is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present utility model without departing from its scope should be within the protection scope of the present utility model.
Claims
1. A device for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis, characterized in that, It includes an L-shaped base (1), a pair of fixed tubes (2) on the bottom surface of the L-shaped base (1), centrifuge tubes (3) respectively disposed in the fixed tubes (2), and a filter separation assembly disposed at the top of the centrifuge tubes (3); The two centrifuge tubes (3) are used to collect interstitial cells and epithelial cells respectively, and the top of each centrifuge tube (3) is connected to a cap (4); The filtration and separation assembly includes an installation ring (10), a positioning ring (11), a flexible filter screen (13), a ring frame (14), and a positioning mechanism; the installation ring (10) is horizontally arranged, and the positioning ring (11) is arranged around the periphery of the installation ring (10) and the positioning ring (11) is adapted to the top end of the centrifuge tube (3); The flexible filter (13) is located at the bottom of the mounting ring (10), the bottom of the ring frame (14) is connected to the bottom surface of the flexible filter (13), and there is a gap between the top of the ring frame (14) and the mounting ring (10), and the ring frame (14) is limited by the positioning mechanism.
2. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, On the vertical surface of the L-shaped base (1), a pair of limiting plates (7) are provided near the rotation; The bottom of the limiting plate (7) is provided with an arc-shaped limiting groove (8), and the arc of the arc-shaped limiting groove (8) matches the outer edge of the top cover (4).
3. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 2, characterized in that, The L-shaped base (1) has a locking block (9) on its vertical surface directly above the limiting plate (7) for limiting the limiting plate (7).
4. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The outer wall of the mounting ring (10) is horizontally connected to a handle (15).
5. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The inner diameter of the positioning ring (11) is adapted to the outer diameter of the centrifuge tube (3).
6. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The positioning ring (11) has a clearance opening (12) on one side to provide clearance for the top cover (4).
7. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The flexible filter (13) is a deformable filter that can deform in the opposite direction under force, and the pore size of the flexible filter (13) is 30-50μm.
8. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The annular frame (14) has a notch on its side wall for the flow of cell fluid.
9. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, The positioning mechanism includes a set screw (16) threaded through the outer wall of the mounting ring (10), a protrusion (18) fixed to the inner wall of the mounting ring (10), and a circular groove (17) and a recess (19) respectively opened on the outer walls of both sides of the ring frame (14). One end of the set screw (16) can abut against the circular groove (17), and the protrusion (18) can abut against the groove (19).
10. The apparatus for isolating and culturing ectopic epithelial and stromal cells of human adenomyosis according to claim 1, characterized in that, Parallel guide blocks (5) are symmetrically arranged on the vertical surface of the L-shaped base (1); A shield (6) is slidably connected between the parallel guide blocks (5).