Lung cancer organoid culture scaffold and organoid culture method

Abstract

The invention relates to the technical field of lung cancer organoid culture, in particular to a lung cancer organoid culture support which comprises a box-shaped support, a disc is fixedly connected to the position, close to the upper edge, of the inner wall of the box-shaped support, the outer wall of the box-shaped support communicates with an annular shell, and three cylinders are fixedly installed at the upper end of the disc in a penetrating mode; and the inner walls of the three cylinders are fixedly connected with spacer rings. The culture space in each culture dish simulates the normal breathing state, the lung breathing rhythm of the human body is mechanically simulated, culture conditions close to the physiological environment of the human body are constructed for lung cancer organ cells, the bionic degree of cell culture is remarkably improved, and a more real cell model is provided for related research. The culture spaces in the three culture dishes can simulate the state of tachypnea, and the tachypnea breathing amplitude and the normal breathing amplitude simulated by the culture spaces in the three culture dishes can be adjusted, so that the application range is expanded, and the research applicability is improved.

Description

Technical Field

[0001] This invention relates to the field of lung cancer organoid culture technology, specifically to a lung cancer organoid culture scaffold and organoid culture method. Background Technology

[0002] Lung cancer organoids, as three-dimensional cell models derived from tumor tissue of lung cancer patients, can highly replicate the characteristics of the primary tumor and have outstanding value in lung cancer research, personalized treatment, and drug development. They serve as a key carrier bridging basic research and clinical practice. Current mainstream culture devices include commercial CO2 incubators, organoid-specific thermostats, and customized dynamic systems. Their core components often utilize biocompatible medical polymer materials to ensure a sterile and safe culture environment. The core function of these devices is to maintain relevant stable conditions. The standard culture procedure involves processing and separating tumor tissue cells, mixing them with matrix gel, inoculating them into culture dishes, and then placing them in the device for culture. With the development of precision medicine, the requirements for the environmental stability and functional simulation of these devices are constantly increasing.

[0003] However, existing culture methods merely keep the culture dish static, failing to simulate the different respiratory rhythms of the human lung to create culture conditions for lung cancer organoid cells that closely resemble the human physiological environment. They only simulate the basic conditions for survival, but not the core physiological characteristics. As a result, while the organoid model can reproduce the basic structure of the tumor, it loses the key biological behaviors and microenvironmental characteristics of lung cancer in the human body, affecting its core value as a clinical alternative model. Therefore, we propose a lung cancer organoid culture scaffold and organoid culture method. Summary of the Invention

[0004] The purpose of this invention is to provide a lung cancer organoid culture scaffold and an organoid culture method to solve the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a lung cancer organoid culture scaffold, comprising a box-shaped scaffold, wherein a disc is fixedly connected to the inner wall of the box-shaped scaffold near its upper edge, and an annular shell is connected to the outer wall of the box-shaped scaffold; three cylinders are fixedly installed through the upper end of the disc; spacers are fixedly connected to the inner walls of the three cylinders; elastic films are slidably attached to the upper ends of the three spacers; culture dishes are fixedly connected to the upper ends of the three elastic films; positioning mechanisms are provided between the three culture dishes and the three cylinders respectively; a second motor is fixedly installed at the center of the bottom of the box-shaped scaffold; three L-shaped plates fixed together are fixedly connected to the output shaft of the second motor; lifting mechanisms are provided between the three L-shaped plates and the three cylinders respectively; and the three lifting mechanisms are used to intermittently lift the three elastic films.

[0006] Preferably, the positioning mechanism includes multiple clamping columns, which are arranged in a ring array and slidably attached to the outer ring surface of the culture dish. The upper ends of the multiple clamping columns are all fixedly connected to inclined columns, and elastic mechanisms are provided between the multiple clamping columns and the cylinder.

[0007] Preferably, the elastic mechanism includes two guide cylinders, which are symmetrically fixedly connected to the outer wall of the cylinder. A guide rod is slidably inserted into the end of each guide cylinder away from the cylinder. Both guide rods movably pass through the cylinder, and the inner end of each guide rod is fixedly connected to the outer wall of the clamping column. A ring is slidably sleeved on the outer wall of each guide rod. The two rings are slidably fitted onto the inner walls of the two guide cylinders. A second spring is fixedly connected between the outer wall of each ring and the inner wall of each guide cylinder, and the second spring is slidably sleeved on the outer wall of each guide rod.

[0008] Preferably, the lifting mechanism includes a U-shaped block, which is slidably connected to the outer vertical wall of the L-shaped plate. A connecting pipe is fixedly connected to the upper end of the U-shaped block, and a connecting rod is slidably inserted into the interior of the connecting pipe from top to bottom. A carrier plate is fixedly connected to the upper end of the connecting rod, and a magnetic block is fixedly connected to the upper end of the carrier plate. A spring is fixedly connected between the lower end of the connecting rod and the upper end of the U-shaped block. A magnetic pushing assembly is provided between the magnetic block and the elastic film directly above it, and a reciprocating moving assembly is provided between the carrier plate and the annular shell.

[0009] Preferably, the magnetic actuation assembly includes a connecting column, which is slidably inserted into the inner wall of the bottom end of the cylinder. A second magnetic block is fixedly connected to the lower end of the connecting column, and a top block is fixedly connected to the upper end of the connecting column. A first spring is fixedly connected between the lower end of the top block and the inner bottom end of the cylinder. The first spring is slidably sleeved on the outer wall of the connecting column. The upper end face of the top block is close to the lower end face of the elastic film. The centers of the top block, the elastic film, the first magnetic block, and the second magnetic block are located on the same straight line.

[0010] Preferably, the reciprocating moving component includes an L-shaped block, which is slidably connected to the inner bottom of the annular shell. A motor is fixedly mounted on the upper end of the L-shaped block, and a support frame is fixedly connected to the vertical wall of the L-shaped block. A short rod is fixedly connected to the output shaft end of the motor. A long rod is rotatably connected to the end of the short rod away from the output shaft of the motor. A guide post is rotatably connected to the end of the long rod away from the short rod. The guide post is slidably inserted into the inner wall of the support frame, and a connecting plate is fixedly connected to the end of the guide post away from the long rod. A triangular block is slidably connected to the wall surface of the connecting plate away from the guide post. An elastic connecting component is provided between the triangular block and the U-shaped block.

[0011] Preferably, the elastic connection component includes a cylindrical groove, which is formed on the end of the U-shaped block away from the L-shaped plate. A spring four is fixedly connected to the groove wall of the cylindrical groove near the L-shaped plate. An L-shaped rod is fixedly connected to the end of the spring four away from the L-shaped plate. The horizontal part of the L-shaped rod is slidably engaged with the cylindrical groove, and the upper end of the L-shaped rod is fixedly connected to the lower end of the triangular block.

[0012] Preferably, an electric push rod is fixedly installed at the lower center of the disc, and a Y-shaped frame is fixedly connected to the telescopic shaft end of the electric push rod. The three ends of the Y-shaped frame are respectively fixedly connected to the upper edge of the outer wall of the three connecting pipes.

[0013] This invention also discloses a method for culturing organoids using a lung cancer organoid culture scaffold, comprising the following steps: S1, Sample processing and cell suspension preparation: lung cancer tissue samples are taken, and single cells are separated after aseptic treatment. The cells are then mixed with matrix gel containing growth factors to prepare a uniform cell suspension. S2, Inoculation of culture dishes: Inoculate the cell suspension into a special culture dish with an elastic film structure at the bottom, ensuring uniform inoculation, and then seal the dish. S3, Placing the culture dish into the box: Place the inoculated culture dish into the spacer rings inside the three cylinders in sequence, and the positioning mechanism can quickly and easily fix the culture dish in place. S4, through the cooperation of motor 2, L-shaped plate and lifting mechanism, allows the elastic film to be gently squeezed back and forth, thus simulating human breathing through repeated movements. The frequency and degree of the back and forth squeezing can be adjusted, thereby mechanically simulating different respiratory rhythms of the human lungs. This provides lung cancer organoid cells with culture conditions that closely resemble the human physiological environment, significantly improving the biomimicry and growth stability of cell culture, and providing a more realistic cell model for related research.

[0014] Compared with the prior art, the beneficial effects of the present invention are: 1. Through the cooperation of box-shaped support, disc, annular shell, cylinder, septum, elastic film, culture dish, positioning mechanism, motor, L-shaped plate and lifting mechanism, each culture dish can be easily fixed, and the culture space in each culture dish simulates normal breathing. By mechanically simulating the breathing rhythm of human lungs, culture conditions that are close to the physiological environment of the human body are constructed for lung cancer organoid cells, which significantly improves the biomimicry of cell culture and provides a more realistic cell model for related research.

[0015] 2. The culture spaces within the three petri dishes can also simulate a state of rapid breathing, thereby expanding the scope of application and research applicability. Furthermore, the amplitude of rapid breathing and normal breathing simulated in the culture spaces within the three petri dishes can be adjusted, further expanding the scope of application and research applicability. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 This is a cross-sectional view of the box-shaped support of the present invention; Figure 3 This is a cross-sectional view of the box-shaped support, culture dish, septum ring, cylinder and connecting tube of the present invention; Figure 4 For the present invention Figure 3 Enlarged view of the structure at point A in the middle; Figure 5 For the present invention Figure 3 Enlarged view of the structure at point B in the middle; Figure 6 This is a diagram illustrating the motor, guide post, triangular block, connecting plate, mounting plate, short rod, and long rod of the present invention. Figure 7 This is a cross-sectional view of the petri dish and cylinder of the present invention; Figure 8 This is a schematic diagram of the internal structure of the box-shaped support of the present invention.

[0017] The attached diagram lists the components represented by each number as follows: 1. Petri dish; 2. Disc; 3. Cylinder; 4. Box-shaped support; 5. Motor 1; 6. Connecting tube; 7. Magnetic block 1; 8. Y-shaped frame; 9. Spacer ring; 10. Top block; 11. Spring 1; 12. Connecting column; 13. Guide column; 14. Triangular block; 15. Connecting plate; 16. L-shaped rod; 17. Connecting rod; 18. L-shaped plate; 19. Motor 2; 20. Magnetic block 2; 21. U-shaped block; 22. Electric push rod; 23. Carrier plate; 24. Guide rod; 25. Guide cylinder; 26. Spring 2; 27. Ring; 28. Clamping column; 29. ​​Inclined column; 30. Spring 3; 31. Spring 4; 32. L-shaped block; 33. Short rod; 34. Long rod; 35. Elastic membrane; 36. Columnar groove; 37. Annular shell; 38. Support frame. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] This invention provides a technical solution: such as Figure 1 - Figure 8The lung cancer organoid culture scaffold shown includes a box-shaped scaffold 4. A disc 2 is fixedly connected to the inner wall of the box-shaped scaffold 4 near its upper edge, and an annular shell 37 is connected to the outer wall of the box-shaped scaffold 4. Three cylinders 3 are fixedly installed through the upper end of the disc 2. Spacer rings 9 are fixedly connected to the inner walls of the three cylinders 3. Elastic films 35 are slidably attached to the upper ends of the three spacer rings 9. Culture dishes 1 are fixedly connected to the upper ends of the three elastic films 35. Positioning mechanisms are provided between the three culture dishes 1 and the three cylinders 3 respectively. A motor 2 19 is fixedly installed at the center of the bottom of the box-shaped scaffold 4. Three L-shaped plates fixed together are fixedly connected to the output shaft of the motor 2 19. 18. Three L-shaped plates 18 are respectively provided with lifting mechanisms between them and three cylinders 3. The three lifting mechanisms are used to intermittently lift the three elastic films 35. Specifically, by setting a positioning mechanism, the three culture dishes 1 can be fixed in the three cylinders 3 respectively. Through the cooperation between the motor 19, L-shaped plates 18, annular shell 37 and lifting mechanisms, each elastic film 35 can be reciprocated and squeezed, so that the culture space of the three culture dishes 1 can simulate the normal breathing state, thereby simulating the breathing rhythm of human lungs, constructing culture conditions that are close to the physiological environment of human lung cancer organoid cells, significantly improving the biomimicry of cell culture, and providing a more realistic cell model for related research.

[0020] The positioning mechanism includes multiple clamping columns 28, which are arranged in a ring array and slidably attached to the outer ring surface of the culture dish 1. Each of the multiple clamping columns 28 has an inclined column 29 fixedly connected to its upper end, and each of the multiple clamping columns 28 has an elastic mechanism between it and the cylinder 3. The elastic mechanism includes two guide cylinders 25, which are symmetrically fixed to the outer wall of the cylinder 3. Guide rods 24 are slidably inserted into the ends of the two guide cylinders 25 away from the cylinder 3. Both guide rods 24 movably pass through the cylinder 3, and their inner ends are fixedly connected to the outer wall of the clamping column 28. Rings 27 are slidably fitted onto the outer walls of the two guide rods 24, respectively, and are slidably fitted against the inner walls of the two guide cylinders 25. Springs 26 are fixedly connected between the outer surfaces of the two rings 27 and the inner walls of the two guide cylinders 25, respectively, and are slidably fitted onto the outer walls of the two guide rods 24. Specifically, by setting up the cooperation between the positioning mechanism and the elastic mechanism, three culture dishes 1 can be quickly and easily fixed inside the three cylinders 3, making operation convenient.

[0021] The lifting mechanism includes a U-shaped block 21, which is slidably connected to the outer vertical wall of the L-shaped plate 18. A connecting pipe 6 is fixedly connected to the upper end of the U-shaped block 21. A connecting rod 17 is slidably inserted into the interior of the connecting pipe 6 from top to bottom. A carrier plate 23 is fixedly connected to the upper end of the connecting rod 17. A magnetic block 7 is fixedly connected to the upper end of the carrier plate 23. A spring 30 is fixedly connected between the lower end of the connecting rod 17 and the upper end of the U-shaped block 21. A magnetic pushing assembly is provided between the magnetic block 7 and the elastic film 35 directly above it. A reciprocating moving assembly is provided between the carrier plate 23 and the annular shell 37. The magnetic drive assembly includes a connecting column 12, which is slidably inserted into the inner wall of the bottom end of the cylinder 3. A second magnetic block 20 is fixedly connected to the lower end of the connecting column 12, and a top block 10 is fixedly connected to the upper end of the connecting column 12. A first spring 11 is fixedly connected between the lower end of the top block 10 and the inner bottom end of the cylinder 3. The first spring 11 is slidably sleeved on the outer wall of the connecting column 12. The upper end face of the top block 10 is close to the lower end face of the elastic film 35. The centers of the top block 10, the elastic film 35, the first magnetic block 7, and the second magnetic block 20 are located on the same straight line. The reciprocating moving component includes an L-shaped block 32, which is slidably connected to the inner bottom of the annular shell 37. A motor 5 is fixedly installed on the upper end of the L-shaped block 32, and a support frame 38 is fixedly connected to the vertical wall of the L-shaped block 32. A short rod 33 is fixedly connected to the output shaft end of the motor 5. A long rod 34 is rotatably connected to the end of the short rod 33 away from the output shaft of the motor 5. A guide post 13 is rotatably connected to the end of the long rod 34 away from the short rod 33. The guide post 13 is slidably inserted into the inner wall of the support frame 38, and a connecting plate 15 is fixedly connected to the end of the guide post 13 away from the long rod 34. A triangular block 14 is slidably connected to the wall surface of the connecting plate 15 away from the guide post 13. An elastic connecting component is provided between the triangular block 14 and the U-shaped block 21. The elastic connection assembly includes a cylindrical groove 36, which is located on the end of the U-shaped block 21 away from the L-shaped plate 18. A spring 31 is fixedly connected to the groove wall of the cylindrical groove 36 near the L-shaped plate 18. An L-shaped rod 16 is fixedly connected to the end of the spring 31 away from the L-shaped plate 18. The horizontal part of the L-shaped rod 16 slides with the cylindrical groove 36, and the upper end of the L-shaped rod 16 is fixedly connected to the lower end of the triangular block 14. Specifically, through the cooperation of the lifting mechanism, the magnetic pushing assembly, the reciprocating moving assembly, and the elastic connection assembly, the culture space in the three culture dishes 1 can simulate a state of rapid breathing, thereby expanding the application range and research applicability.

[0022] An electric push rod 22 is fixedly installed at the lower center of the disc 2. A Y-shaped frame 8 is fixedly connected to the telescopic shaft end of the electric push rod 22. The three ends of the Y-shaped frame 8 are fixedly connected to the upper edge of the outer wall of the three connecting tubes 6, respectively. Specifically, by setting the electric push rod 22 and the Y-shaped frame 8, the simulated rapid breathing amplitude and normal breathing amplitude in the culture space of the three culture dishes 1 can be adjusted, thereby further expanding the application scope and research applicability.

[0023] Working principle: First, the petri dish 1 is installed inside the cylinder 3. The petri dish 1 is then inserted from top to bottom along the direction of multiple inclined columns 29. The bottom edge of the petri dish 1 will press against the inclined surfaces of the multiple inclined columns 29, causing each inclined column 29 to move its connected clamping column 28 away from the center of the cylinder 3. Each clamping column 28 will also cause the two connected guide rods 24 to slide outward within the cylinder 3 and the corresponding guide tube 25. Each guide rod 24 will also cause the connected ring 27 to compress the corresponding spring 26 until the petri dish 1... The elastic film 35 at the bottom of the culture dish 1 comes into contact with the spacer ring 9 inside the cylinder 3. At this time, the culture dish 1 is released, and under the force of multiple springs 26, multiple clamping columns 28 can clamp and fix the culture dish 1. Through the setting of multiple inclined columns 29, culture dishes 1 of different sizes, all with elastic films 35 at the bottom, can be installed in this way. The center of the culture dish 1, the elastic film 35, and the cylinder 3 are all on the same line. Then, fix other culture dishes 1 in the same way. The specific state of the three culture dishes 1 at this time is as described above. Figure 2 - Figure 4 .

[0024] Then, motor 219 is started to drive the three L-shaped plates 18 to rotate. Each L-shaped plate 18 will drive its connected U-shaped block 21, connecting pipe 6, connecting rod 17, carrier plate 23, magnetic block 1 7, and spring 30 to rotate together. Each U-shaped block 21 can also drive its connected L-shaped rod 16, triangular block 14, connecting plate 15, support frame 38, L-shaped block 32, motor 1 5, short rod 33, long rod 34, and guide post 13 to rotate together. The L-shaped block 32 will rotate at the bottom inner side of the annular shell 37 until each magnetic block 1 7 has rotated to below its corresponding magnetic block 20. At this time, the state of the above-mentioned components is as follows. Figure 3 As shown. Since the magnetic poles of magnetic block 7 and magnetic block 20 are opposite, this will drive each magnetic block 20 upwards. Each magnetic block 20 will cause the connecting post 12 to slide upwards within its corresponding cylinder 3. Furthermore, each connecting post 12 will cause its connected top block 10 to move upwards, and the top block 10 will stretch its corresponding spring 11. The upward movement of each top block 10 will exert a certain amount of pressure on its corresponding elastic membrane 35, causing the elastic membrane 35 to deform. When each magnetic block 20 has rotated away from its corresponding magnetic block 20, in... Under the force of spring 11, each magnetic block 20 and each top block 10 will move down and reset, and each elastic membrane 35 will also recover. This process is repeated, so that each elastic membrane 35 can be repeatedly deformed and recovered, thus simulating the normal breathing state in the culture space of each culture dish 1. By mechanically simulating the breathing rhythm of human lungs, culture conditions that are close to the physiological environment of the human body are constructed for lung cancer organoid cells, significantly improving the biomimicry of cell culture and providing a more realistic cell model for related research.

[0025] Furthermore, when each magnetic block 7 has rotated to a position below its corresponding magnetic block 20, each motor 5 is activated, referring to... Figure 6 Each motor 5 drives the short rod 33 to rotate, and each short rod 33 drives its connected long rod 34 to swing back and forth. Each long rod 34 drives its connected guide post 13 to slide back and forth within the corresponding support frame 38. Each guide post 13 can drive the connected connecting plate 15 and the triangular block 14 to move back and forth. Each triangular block 14 is constantly moving towards and away from the corresponding carrier plate 23 and reciprocating. When each triangular block 14 moves towards the carrier plate 23, the inclined surface of each triangular block 14 will press against the lower edge of the corresponding carrier plate 23, causing it to move upward. Each carrier plate 23 will also drive the corresponding magnet As block 14 and connecting rod 17 move upward, each connecting rod 17 slides upward within its corresponding connecting tube 6, stretching its corresponding spring 30. When each triangular block 14 moves away from its corresponding carrier plate 23, under the force of spring 30, all carrier plates 23, magnetic block 14, and connecting rod 17 automatically move downward. This process repeats, allowing each magnetic block 14 to move up and down repeatedly, causing each magnetic block 20 and top block 10 to move up and down as well. This allows each elastic membrane 35 to deform and recover more quickly, simulating a state of rapid breathing in the culture space, thus expanding the application scope and research applicability. It should be noted that when each triangular block 14 moves towards its corresponding carrier plate 23, it also causes its connected L-shaped rod 16 to slide inward in its corresponding cylindrical groove 36 and compress spring 4 31. Conversely, it slides outward in the cylindrical groove 36, and spring 4 31 recovers.

[0026] Furthermore, when the three L-shaped plates 18 rotate, causing the three connecting pipes 6 to rotate together, the Y-shaped frame 8 will also rotate at the telescopic shaft end of the electric push rod 22. Activating the electric push rod 22 drives the Y-shaped frame 8 downwards, thereby causing the three connecting pipes 6 to move downwards as well. This causes all the U-shaped blocks 21, connecting rods 17, springs 30, carrier plates 23, magnetic blocks 7, L-shaped rods 16, and triangular blocks 14 to move downwards. Each triangular block 14 will slide down on its corresponding connecting plate 15 surface. Adjusting the height of each magnetic block 7 ensures that each triangular block 14 can continue to push the carrier plate 23. That is, after the height of each magnetic block 7 is lowered, driving each triangular block 14 to move back and forth causes the corresponding carrier plate 23 and magnetic block 7 to move up and down repeatedly. Because the height of magnetic block 7 is lowered, the magnetic thrust on magnetic block 20 is reduced, thereby adjusting the reciprocating deformation of the elastic film 35. Meanwhile, when the first magnetic block 7 is rotated normally to the position below the corresponding second magnetic block 20 without being affected by the triangular block 14, the deformation of the elastic film 35 will also decrease, thereby adjusting the amplitude of rapid breathing and normal breathing in the culture space simulation.

[0027] It should be noted that the sliding connection between the L-shaped plate 18 and the U-shaped block 21 is a sliding fit between a T-shaped slider and a T-shaped groove. Specifically, the T-shaped slider is connected to the U-shaped block 21, and the T-shaped groove is formed on the L-shaped plate 18. Similarly, the sliding connection between the connecting plate 15 and the triangular block 14 is also a sliding fit between a T-shaped slider and a T-shaped groove. Specifically, the T-shaped slider is connected to the triangular block 14, and the T-shaped groove is formed on the connecting plate 15. This sliding fit between the T-shaped slider and the T-shaped groove is known prior art and will not be elaborated further. Furthermore, the sliding connection between the L-shaped block 32 and the inner bottom end of the annular shell 37 is a sliding fit between an annular slider and an annular groove. Both the annular slider and the annular groove have T-shaped longitudinal sections. The annular slider is connected to the bottom end of the L-shaped block 32, and the annular groove is formed on the inner bottom end of the annular shell 37. This is also known prior art and will not be elaborated further.

[0028] This invention also discloses a method for culturing organoids using a lung cancer organoid culture scaffold, comprising the following steps: S1, Sample processing and cell suspension preparation: lung cancer tissue samples are taken, and single cells are separated after aseptic treatment. The cells are then mixed with matrix gel containing growth factors to prepare a uniform cell suspension. S2, Inoculation of culture dish 1: Inoculate the cell suspension into a special culture dish 1 with an elastic film 35 structure at the bottom, ensuring uniform inoculation, and then cover and seal it. S3, the culture dish 1 is placed in the box. The inoculated culture dish 1 is placed on the spacer ring 9 inside the three cylinders 3 in sequence, and the culture dish 1 can be quickly and easily fixed by the positioning mechanism. S4, through the cooperation of motor 19, L-shaped plate 18 and lifting mechanism, allows the elastic film 35 to be slightly reciprocated, thereby repeatedly simulating human breathing. The frequency and degree of reciprocation can be adjusted, thus mechanically simulating different respiratory rhythms of the human lungs. This provides lung cancer organoid cells with culture conditions that closely resemble the human physiological environment, significantly improving the biomimicry and growth stability of cell culture, and providing a more realistic cell model for related research.

[0029] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0030] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A lung cancer organoid culture scaffold, comprising a box-shaped scaffold (4), characterized in that: The inner wall of the box-shaped support (4) is fixedly connected to a disc (2) near the upper edge, and the outer wall of the box-shaped support (4) is connected to an annular shell (37). Three cylinders (3) are fixedly installed through the upper end of the disc (2). The inner walls of the three cylinders (3) are fixedly connected to spacers (9). The upper ends of the three spacers (9) are slidably fitted with elastic films (35). The upper ends of the three elastic films (35) are fixedly connected to petri dishes (1). The three petri dishes (1) are respectively provided with positioning mechanisms between them and the three cylinders (3). The inner bottom center of the box-shaped support (4) is fixedly installed with a second motor (19). The output shaft end of the second motor (19) is fixedly connected to three L-shaped plates (18) that are fixed together. The three L-shaped plates (18) are respectively provided with lifting mechanisms between them and the three cylinders (3). The three lifting mechanisms are respectively used to intermittently lift the three elastic films (35).

2. The lung cancer organoid culture scaffold according to claim 1, characterized in that: The positioning mechanism includes multiple clamping columns (28), which are arranged in a ring array and slidably attached to the outer ring surface of the culture dish (1). The upper ends of the multiple clamping columns (28) are all fixedly connected to inclined columns (29), and elastic mechanisms are provided between the multiple clamping columns (28) and the cylinder (3).

3. The lung cancer organoid culture scaffold according to claim 2, characterized in that: The elastic mechanism includes two guide cylinders (25), which are symmetrically fixedly connected to the outer wall of the cylinder (3). The ends of the two guide cylinders (25) away from the cylinder (3) are slidably inserted with guide rods (24). The two guide rods (24) are movably inserted through the cylinder (3). The ends of the two guide rods (24) located inside the cylinder (3) are fixedly connected to the outer wall of the clamping column (28). The outer walls of the two guide rods (24) are slidably fitted with rings (27). The two rings (27) are slidably fitted onto the inner walls of the two guide cylinders (25). The walls of the two rings (27) away from the cylinder (3) are fixedly connected to the inner walls of the two guide cylinders (25) with springs (26). The two springs (26) are slidably fitted onto the outer walls of the two guide rods (24).

4. The lung cancer organoid culture scaffold according to claim 1, characterized in that: The lifting mechanism includes a U-shaped block (21), which is slidably connected to the outer vertical wall of the L-shaped plate (18). A connecting pipe (6) is fixedly connected to the upper end of the U-shaped block (21). A connecting rod (17) is slidably inserted into the inside of the connecting pipe (6) from top to bottom. A carrier plate (23) is fixedly connected to the upper end of the connecting rod (17). A magnetic block (7) is fixedly connected to the upper end of the carrier plate (23). A spring (30) is fixedly connected between the lower end of the connecting rod (17) and the upper end of the U-shaped block (21). A magnetic pushing component is provided between the magnetic block (7) and the elastic film (35) directly above it. A reciprocating moving component is provided between the carrier plate (23) and the annular shell (37).

5. The lung cancer organoid culture scaffold according to claim 4, characterized in that: The magnetic drive assembly includes a connecting column (12), which is slidably inserted into the inner wall of the bottom end of the cylinder (3). A magnetic block two (20) is fixedly connected to the lower end of the connecting column (12), and a top block (10) is fixedly connected to the upper end of the connecting column (12). A spring one (11) is fixedly connected between the lower end of the top block (10) and the inner bottom end of the cylinder (3). The spring one (11) is slidably sleeved on the outer wall of the connecting column (12). The upper end face of the top block (10) is close to the lower end face of the elastic film (35). The centers of the top block (10), the elastic film (35), the magnetic block one (7), and the magnetic block two (20) are located on the same straight line.

6. The lung cancer organoid culture scaffold according to claim 4, characterized in that: The reciprocating moving component includes an L-shaped block (32), which is slidably connected to the inner bottom of the annular shell (37). A motor (5) is fixedly installed on the upper end of the L-shaped block (32), and a support frame (38) is fixedly connected to the vertical wall of the L-shaped block (32). A short rod (33) is fixedly connected to the output shaft end of the motor (5). A long rod (34) is rotatably connected to the end of the short rod (33) away from the output shaft of the motor (5). A guide post (13) is rotatably connected to the end of the long rod (34) away from the short rod (33). The guide post (13) is slidably inserted into the inner wall of the support frame (38), and a connecting plate (15) is fixedly connected to the end of the guide post (13) away from the long rod (34). A triangular block (14) is slidably connected to the wall surface of the connecting plate (15) away from the guide post (13). An elastic connecting component is provided between the triangular block (14) and the U-shaped block (21).

7. The lung cancer organoid culture scaffold according to claim 6, characterized in that: The elastic connection assembly includes a cylindrical groove (36), which is opened on the end of the U-shaped block (21) away from the L-shaped plate (18). A spring four (31) is fixedly connected to the groove wall of the cylindrical groove (36) near the L-shaped plate (18). An L-shaped rod (16) is fixedly connected to the end of the spring four (31) away from the L-shaped plate (18). The horizontal part of the L-shaped rod (16) is slidably engaged with the cylindrical groove (36), and the upper end of the L-shaped rod (16) is fixedly connected to the lower end of the triangular block (14).

8. The lung cancer organoid culture scaffold according to claim 1, characterized in that: An electric push rod (22) is fixedly installed at the lower middle part of the disc (2). A Y-shaped frame (8) is fixedly connected to the telescopic shaft end of the electric push rod (22). The three ends of the Y-shaped frame (8) are fixedly connected to the upper edge of the outer wall of the three connecting pipes (6).

9. A method for culturing lung cancer organoids using a scaffold, applied to the lung cancer organoid scaffold described in any one of claims 1-8, characterized in that, Includes the following steps: S1, Sample processing and cell suspension preparation: lung cancer tissue samples are taken, and single cells are separated after aseptic treatment. The cells are then mixed with matrix gel containing growth factors to prepare a uniform cell suspension. S2, Inoculation of culture dish (1): Inoculate the cell suspension into a special culture dish (1) with an elastic film (35) structure at the bottom, ensuring uniform inoculation, and then seal the dish. S3, the culture dish (1) is placed in the box. The inoculated culture dish (1) is placed on the spacer ring (9) inside the three cylinders (3) in sequence, and the culture dish (1) can be quickly and easily fixed by the positioning mechanism. S4, through the cooperation of motor 2 (19), L-shaped plate (18) and lifting mechanism, the elastic film (35) can be slightly squeezed back and forth, thus repeatedly simulating human breathing. The frequency and degree of the back and forth squeezing can be adjusted, thereby mechanically simulating different breathing rhythms of the human lungs, constructing culture conditions close to the human physiological environment for lung cancer organoid cells, significantly improving the biomimicry and growth stability of cell culture, and providing a more realistic cell model for related research.

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