Cell pulsation culture device, drug screening method and manufacturing process

Abstract

The invention discloses a cell pulsation culture device, a drug screening method and a manufacturing process. The cell pulsation culture device comprises a base, a first connecting plate, a second connecting plate and a reaction bin, the first connecting plate is connected with the base, the second connecting plate is slidably connected with the base, the second connecting plate is configured to be close to or away from the first connecting plate in the horizontal direction, and the first connecting plate and the second connecting plate are provided with a plurality of first mounting grooves and second mounting grooves correspondingly. Two ends of the reaction bin are respectively arranged in the first mounting groove and the second mounting groove, and the reaction bin is used for accommodating a target sample. Wherein the reaction bin has elasticity, and the second connecting plate can be configured to be close to or far away from the first connecting plate periodically along preset parameters, so that the reaction bin provides at least one waveform of rising branch, dominant wave, tidal wave, falling strait and dicrotic wave for the target sample. According to the invention, a plurality of complex waveforms can be simulated, homogeneous mechanical stimulation of a multi-channel target sample is realized, the experiment efficiency is high, and the reliability is high.

Description

Technical Field

[0001] This invention relates to the fields of cell mechanics and drug screening technology, and in particular to a cell pulsation culture device, a drug screening method, and a manufacturing process. Background Technology

[0002] In living organisms, blood vessel walls are constantly subjected to periodic mechanical stresses caused by the heartbeat. This pulsating stretching not only maintains vascular homeostasis but also profoundly affects the proliferation, differentiation, and drug response of vascular endothelial cells and smooth muscle cells. However, most traditional biological experiments are conducted in static culture dishes, lacking mechanical stimulation that matches the physiological environment. This leads to a large number of active compounds sensitive to mechanical signals being missed in early screening, increasing the time and cost of new drug development.

[0003] While some existing technologies have developed mechanical devices capable of stretching cells, they generally suffer from low throughput, complex structures, cumbersome clamping, and most can only perform simple stretching operations. Furthermore, commercially available equipment is expensive and bulky, making it impossible to place the entire device into an incubator or microscope stage for long-term, real-time observation. Summary of the Invention

[0004] The main objective of this invention is to propose a cell pulsation culture device, a drug screening method, and a manufacturing process that can simulate multiple complex waveforms and achieve homogeneous mechanical stimulation of multi-channel target samples, resulting in high experimental efficiency and reliability.

[0005] To achieve the above objectives, some embodiments of the present invention provide a cell pulsation culture device, comprising: Base The first connecting plate, connecting base, has multiple spaced first mounting slots along the vertical direction; The second connecting plate and the sliding connecting base are configured to be able to approach or move away from the first connecting plate in the horizontal direction, and in the vertical direction, the second connecting plate is provided with a plurality of second mounting slots corresponding to the first mounting slot. The reaction chamber has two ends that are relatively distributed in the horizontal direction. One end of the reaction chamber is located in the first mounting slot, and the other end of the reaction chamber is located in the second mounting slot. The reaction chamber is used to contain the target sample. The reaction chamber is elastic, and the second connecting plate can be configured to periodically move closer to or further away from the first connecting plate along preset parameters, so that the reaction chamber provides the target sample with at least one waveform among the rising support, main wave, tidal wave, descending isthmus, and diabetic wave.

[0006] In some embodiments, the cell pulsation culture device further includes a third connecting plate, which is connected to the base. The third connecting plate is located on the side of the second connecting plate away from the first connecting plate. In the vertical direction, the third connecting plate is provided with a plurality of spaced third mounting slots. The side of the second connecting plate facing the third connecting plate is provided with a plurality of fourth mounting slots corresponding to the third mounting slots. The third mounting slots and the fourth mounting slots cooperate to install the reaction chamber. The second connecting plate is configured to be adjustable in the horizontal direction to the distance from both the first and third connecting plates.

[0007] In some embodiments, the preset parameters include at least the average speed of the second connecting plate, the acceleration of the second connecting plate, and the distance the second connecting plate moves.

[0008] In some embodiments, the cell pulsation culture device includes a drive motor and a lead screw, one end of which is connected to the drive motor, and the other end of which is connected to a second connecting plate. The drive motor drives the lead screw to move according to preset parameters; and / or, The cell pulsation culture device includes a control unit, which is configured to control the drive motor to operate according to preset parameters.

[0009] In some embodiments, the reaction chamber includes a main body and two pins. The main body defines a receiving chamber for receiving a target sample. Along the direction from the second connecting plate to the first connecting plate, the two pins are respectively inserted at both ends of the main body and are respectively recessed into a first mounting groove and a second mounting groove.

[0010] In some embodiments, the main body includes a central region and an edge region surrounding the central region. The central region is recessed to form a receiving chamber. Along the direction from the second connecting plate to the first connecting plate, the size of the edge region is larger than the size of the central region. Along the direction perpendicular to the direction from the second connecting plate to the first connecting plate, the size of the edge region is larger than the size of the central region.

[0011] In some embodiments, the wall thickness of the reaction chamber ranges from 4mm to 8mm, the length of the reaction chamber ranges from 40mm to 60mm, and the width of the reaction chamber ranges from 15mm to 30mm.

[0012] An embodiment of the second aspect of this application provides a drug screening method, including the cell pulsating culture apparatus described above, characterized in that the drug screening method includes: S101: The same cells were seeded in multiple reaction chambers and different candidate substances were added; S103: Select the waveform parameters for simulating pathological pulsations and apply pulsation stretching; S105: Detects indicators of calcium deposition, apoptosis, or phenotypic transformation in cells within each reaction chamber; S107: Compare the differences in indicators to screen for active substances that can antagonize the pathological pulsation effect.

[0013] An embodiment of the third aspect of this application provides a manufacturing process, characterized in that the manufacturing process for the reaction chamber used in any of the above-mentioned cell pulsating culture devices includes: S201: A concave mold structure is formed by photopolymerization 3D printing, and the concave mold structure corresponds to the outer surface of the reaction chamber; S203: The concave mold is irradiated with ultraviolet light for 2.5h~3.5h, and then placed in an oven at 65℃~75℃ for 4.5h~6.5h to eliminate the leached phosphine oxide photoinitiator in the structure; S205: Degassed liquid polydimethylsiloxane is poured into a concave mold and vacuum degassed; after curing at 65℃~75℃ for 1.5h~2.5h, it is demolded to obtain the elastic reaction chamber body; S207: Insert pins into both ends of the reaction chamber and seal them to obtain the finished reaction chamber.

[0014] In some embodiments, after step S207, the following step is further included: The reaction chamber body was cleaned with anhydrous ethanol, followed by ultrasonic cleaning for 15 to 25 minutes to remove surface residues. After cleaning, the reaction chamber is immersed in 70%~75% alcohol for 0.8h~1.2h, then air-dried and sterilized by ultraviolet light for 2.5h~3.5h. After rinsing with sterile PBS and air-drying again, the reaction chamber was pre-washed with cell culture medium to complete sterilization and pre-equilibration.

[0015] According to the above embodiments, the beneficial effects of the present invention are: An embodiment of the first aspect of this application provides a cell pulsation culture device, including a base, a first connecting plate, a second connecting plate, and a reaction chamber. The first connecting plate is connected to the base and has a plurality of spaced-apart first mounting slots along the vertical direction. The second connecting plate is slidably connected to the base and is configured to move closer to or further away from the first connecting plate in the horizontal direction. In the vertical direction, the second connecting plate has a plurality of second mounting slots corresponding to the first mounting slots. The reaction chamber has two ends distributed opposite each other in the horizontal direction. One end of the reaction chamber is located in the first mounting slot, and the other end is located in the second mounting slot. The reaction chamber is used to contain a target sample. The reaction chamber is elastic, and the second connecting plate can be configured to periodically move closer to or further away from the first connecting plate along preset parameters, so that the reaction chamber provides the target sample with at least one waveform selected from the following: rising wave, main wave, tidal wave, descending wave, and dicrotic wave.

[0016] Specifically, the cell pulsatile culture device achieves physiological-level pulsatile stretching of target samples through the coordinated design of a base, a first connecting plate, a second connecting plate, and an elastic reaction chamber. The mounting slots on the first and second connecting plates allow for rapid and reliable positioning and fixation of the reaction chamber ends, enabling synchronous deformation with the reciprocating motion of the second connecting plate without the need for additional clamps. During periodic axial stretching, the elastic reaction chamber can completely reproduce a composite waveform including the ascending limb, main wave, tidal wave, descending isthmus, and dicrotic wave, providing the sample with a dynamic mechanical environment consistent with in vivo blood vessels. Since the waveform is directly driven by preset parameters, researchers can freely set the motion curve for normal or pathological states, improving the physiological relevance of mechanical stimulation. Furthermore, the array-like arrangement of the mounting slots supports parallel loading of multiple reaction chambers, forming a multi-layered tower array, such as 96-channel or 18-channel arrays, enabling homogeneous pulsatile stimulation of batches of samples in a single operation. This further expands throughput within the limited volume of the incubator, significantly improving experimental efficiency and reducing operational errors, facilitating high-throughput drug combination screening, individual difference testing, and clinical drug screening. The device has a simple overall structure and is easy to assemble and disassemble. It can be moved as a whole into an incubator or microscope platform to achieve real-time observation and intervention under continuous force stimulation, providing a highly controllable and easy-to-use hardware platform for cell mechanics, tissue engineering and disease mechanism research.

[0017] In summary, this application, through a high-throughput cell pulsatile culture device, enables simultaneous experiments on the same batch of target samples, resulting in more accurate and stable data sets. Combined with the motion settings of the second connecting plate, the cell pulsatile culture device can simultaneously simulate multiple waveforms across all reaction chambers to match the actual environment, further improving the reliability and analyzability of the experiments. This application is small in size, can be placed on various microscopic platforms for observation, and combines the advantages of low production cost with high detection efficiency and high reliability, making it suitable for a wide range of applications.

[0018] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention 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 some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0020] Figure 1 This is a three-dimensional structural schematic diagram of the cell pulsating culture device as viewed from a first perspective in one embodiment of the present invention. Figure 2This is a three-dimensional structural schematic diagram of the cell pulsating culture device as viewed from a second perspective in one embodiment of the present invention. Figure 3 This is a three-dimensional structural diagram of the cell pulsating culture device as viewed from a third-person perspective in one embodiment of the present invention. Figure 4 This is a schematic diagram of a partial explosion structure of a cell pulsating culture device in one embodiment of the present invention; Figure 5 This is a three-dimensional structural diagram of the first connecting plate in one embodiment of the present invention; Figure 6 This is a three-dimensional structural diagram of the second connecting plate in one embodiment of the present invention; Figure 7 This is a three-dimensional structural diagram of the reaction chamber in one embodiment of the present invention; Figure 8 This is an image of alizarin red staining of vascular smooth muscle cells under cyclic pulsation culture conditions at different time points in one embodiment of the present invention; Figure 9 Alizarin Red staining images of six typical Chinese herbal extracts in one embodiment of the present invention, demonstrating their inhibitory effect on VSMC calcification; Figure 10 This is a flowchart of a drug screening method in one embodiment of the present invention; Figure 11 This is a flowchart of the manufacturing process in one embodiment of the present invention.

[0021] Explanation of icon numbers: Base 100; First connecting plate 200; First mounting slot 210; Second connecting plate 300; Second mounting slot 310; Fourth mounting slot 320; Reaction chamber 400; Main body 410; Central region 411; Edge region 412; Pin 420; Third connecting plate 500; Third mounting slot 510; Motor 600.

[0022] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0023] 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 a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0024] It should be noted that if the embodiments of the present invention involve directional indicators (such as up, down, left, right, front, back, etc.), the directional indicators are only used to explain the relative positional relationship and movement of the components in a specific posture. If the specific posture changes, the directional indicators will also change accordingly.

[0025] Furthermore, if the embodiments of this invention involve descriptions such as "first" or "second," these descriptions are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of those features. Additionally, the use of "and / or," "and / or," or "and / or" throughout the text implies three parallel solutions. For example, "A and / or B" includes solution A, solution B, or a solution where both A and B are satisfied simultaneously. Furthermore, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or impossible to implement, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by this invention.

[0026] The following is for reference. Figures 1 to 11 This describes the cell pulsation culture apparatus, drug screening method, and manufacturing process according to embodiments of the present invention. (Refer to...) Figures 1 to 7 An embodiment of the first aspect of this application provides a cell pulsatile culture device, including a base 100, a first connecting plate 200, a second connecting plate 300, and a reaction chamber 400. The first connecting plate 200 is connected to the base 100 and, in the vertical direction, has a plurality of spaced-apart first mounting grooves 210. The second connecting plate 300 is slidably connected to the base 100 and is configured to move horizontally towards or away from the first connecting plate 200. In the vertical direction, the second connecting plate 300 has a plurality of second mounting grooves 310 corresponding to the first mounting grooves 210. The reaction chamber 400 has two ends distributed opposite each other in the horizontal direction. One end of the reaction chamber 400 is located in the first mounting groove 210, and the other end is located in the second mounting groove 310. The reaction chamber 400 is used to contain a target sample. The reaction chamber 400 is elastic, and the second connecting plate 300 can be configured to periodically move closer to or further away from the first connecting plate 200 along preset parameters, so that the reaction chamber 400 provides the target sample with at least one waveform among the rising support, main wave, tidal wave, descending mid-slope, and diabetic wave.

[0027] Specifically, the cell pulsatile culture device achieves physiological-level pulsatile stretching of the target sample through the coordinated design of the base 100, the first connecting plate 200, the second connecting plate 300, and the elastic reaction chamber 400. The corresponding mounting slots on the first connecting plate 200 and the second connecting plate 300 allow for rapid and reliable positioning and fixation of both ends of the reaction chamber 400, enabling it to deform synchronously with the reciprocating motion of the second connecting plate 300 without the need for additional clamps. During periodic axial stretching, the elastic reaction chamber 400 can completely reproduce a composite waveform including the ascending limb, main wave, tidal wave, descending isthmus, and dicrotic wave, providing the sample with a dynamic mechanical environment consistent with in vivo blood vessels. Since the waveform is directly driven by preset parameters, researchers can freely set the motion curve for normal or pathological states, improving the physiological relevance of mechanical stimulation. Furthermore, the array-style arrangement of the mounting slots supports parallel loading of up to 400 reaction chambers, forming a multi-layered tower array, such as 96-channel or 18-channel arrays. This allows for the simultaneous homogeneous pulsatile stimulation of batches of samples, further expanding throughput within the limited volume of the incubator. This significantly improves experimental efficiency and reduces operational errors, facilitating high-throughput drug combination screening, individual difference testing, and clinical drug screening. The device has a simple overall structure and is easy to assemble and disassemble. It can be moved entirely into an incubator or microscope platform, enabling real-time observation and intervention under continuous force stimulation. This provides a highly controllable and easy-to-use hardware platform for cell mechanics, tissue engineering, and disease mechanism research.

[0028] In summary, this application, through a high-throughput cell pulsation culture device, enables simultaneous experiments on the same batch of target samples, resulting in more accurate and stable data sets. Combined with the motion settings of the second connecting plate 300, the cell pulsation culture device can simultaneously simulate multiple waveforms in all reaction chambers 400 to match the actual environment, further improving the reliability and analyzability of the experiments. This application is small in size, can be placed on various microscopic platforms for observation, and combines the advantages of low production cost, high detection efficiency, and high reliability, making it suitable for a wide range of applications.

[0029] In some embodiments, the first connecting plate 200 and the second connecting plate 300 can be photocured resin frames, or milled aluminum alloy plates, laser-cut stainless steel plates, or injection-molded polyetheretherketone (PEEK) parts. For example, when the first connecting plate 200 is selected as an integral photocured resin frame, its inner side is arrayed to form the first mounting groove 210. The second connecting plate 300 is also made of photocured resin and is integrally printed with the linear slide rail insert on the base 100. The two are connected to the same ball screw pair through the insert nut. The rotation of the screw can cause the second connecting plate 300 to translate with high precision relative to the first connecting plate 200, thereby causing the elastic reaction chamber 400 stuck between the first mounting groove 210 and the second mounting groove 310 to be axially stretched or rebounded, realizing pulsating waveform loading.

[0030] In some embodiments, the first mounting groove 210 and the second mounting groove 310 may be rectangular notches recessed along the thickness direction of the first connecting plate 200 and the second connecting plate 300. The first mounting groove 210 and the second mounting groove 310 may also be semi-circular clamp-type open grooves, dovetail-shaped slide rail grooves, or blind-hole type insertion sockets. In some embodiments, if the edges of the grooves are made into symmetrical R-shaped chamfers, an elastic latch can be formed, allowing the pins 420 at the ends of the reaction chamber 400 to be automatically clamped after being pressed radially. In some embodiments, the reaction chamber 400 can also be assembled with the first connecting plate 200 and the second connecting plate 300 through additional structures, such as using adhesive magnetic sheets to magnetically fix the ferromagnetic pins 420 to the surfaces of the first connecting plate 200 and the second connecting plate 300. In some embodiments, a quick-release cam lock is used; pulling the handle simultaneously clamps or releases a row of reaction chambers 400, achieving tool-free and rapid clamping.

[0031] In some embodiments, the reaction chamber 400 can be a PDMS rectangular film cavity with stainless steel pins 420 embedded at both ends, or it can be a 3D-printed TPE corrugated tube, a laser-welded TPU microcapsule, an injection-molded PC sheet heat-sealed bag, or an oval capsule made of medical-grade silicone. The reaction chamber 400 needs to be able to produce uniform elongation under axial tension. For example, a 0.2mm thick TPU film can be vacuum-formed into a cavity, and ABS short rods can be heat-fused to both ends to form rigid pins 420. The pins 420 are directly pressed into the mounting groove to complete the assembly, which ensures light transmission for microscopic observation and allows for repeated pulsation without failure within a reasonable strain range.

[0032] In some embodiments, the target sample may be primary vascular smooth muscle cells, cardiomyocytes derived from induced pluripotent stem cells, co-cultured endothelial-smooth muscle bilayer tissue, or mammary organoids, tumor spheres, osteoblast-osteoclast co-culture systems, or decellularized and re-seeded porcine aortic valve tissue, etc. The target sample needs to maintain its viability and function in a controlled pulsatile stretching environment. For example, human iPSC cardiomyocytes can be combined with collagen gel, injected into a PDMS reaction chamber 400, and allowed to stand to gel. This allows for simultaneous recording of contraction rhythm and calcium transients under subsequent pulsatile stimulation, which can be used for screening for drug toxicity or rhythm abnormalities.

[0033] The preset parameters can be the moving average speed, acceleration, and period of proximity to the first connecting plate 200 of the second connecting plate 300. These parameters can simulate waveforms such as a 1Hz sine wave, 10% strain, lasting 4 hours; a 0.2Hz triangular wave, 5% strain, with a 10-minute interval of stretching followed by a 10-minute rest cycle; or a 1.5Hz complex wave with 15% strain and secondary tidal peaks characteristic of hypertension. These parameters are input into the controller via code or a graphical interface, driving the motor 600 to operate according to the corresponding speed and acceleration curves. This allows the reaction chamber 400 to reproduce the mechanical characteristics of arteries in physiological, exercise, or disease states, providing a highly matched mechanical microenvironment for the cells.

[0034] Reference Figures 1 to 6 In some embodiments, the cell pulsation culture device further includes a third connecting plate 500, which is connected to the base 100. The third connecting plate 500 is located on the side of the second connecting plate 300 opposite to the first connecting plate 200. Vertically, the third connecting plate 500 has multiple spaced-apart third mounting slots 510. The second connecting plate 300 has multiple fourth mounting slots 320 corresponding to the third mounting slots 510 on the side facing the third connecting plate 500. The third mounting slots 510 and the fourth mounting slots 320 cooperate to mount the reaction chamber 400. The second connecting plate 300 is configured to be horizontally adjustable to the distances of both the first connecting plate 200 and the third connecting plate 500. Specifically, by providing a synchronously adjustable second connecting plate 300 between the first connecting plate 200 and the third connecting plate 500, the device forms a bilaterally symmetrical loading structure. When the second connecting plate 300 moves closer to one side, the other side moves away simultaneously, thus applying tension and relaxation to the two sets of reaction chambers 400 within the same motion cycle, achieving bidirectional mechanical stimulation without an additional driving source. This layout not only doubles the space utilization rate but also reduces the eccentricity of the slide and mechanical hysteresis through bidirectional force balance, ensuring stable long-term operation and high waveform reproducibility. Researchers only need to clamp once to obtain sample groups with complementary mechanical states under the same temperature and atmosphere, facilitating direct comparison of the differences between the same drug or gene intervention under tension and relaxation states, improving the reliability of research conclusions. The array-type mounting slot design keeps the axial alignment of each reaction chamber 400, eliminating the additional shear force caused by traditional cantilever clamping. The overall structure is simple and compact, and can be placed as a whole into an incubator or microscope stage, enabling real-time observation and intervention of batch samples under bidirectional pulsating stimulation.

[0035] Reference Figures 1 to 4In some embodiments, the preset parameters include at least the average speed of the second connecting plate 300, the acceleration of the second connecting plate 300, and the distance the second connecting plate 300 moves. This design allows researchers to reproduce multiple waveforms on the same device without replacing mechanical components. The adjustable acceleration allows the device to simulate instantaneous pressure surges occurring under pathological conditions, while the continuous variation of the average speed provides a smoother transition between waveform plateaus or decay phases. The set distance ensures that each stretching endpoint can repeatedly return to the same strain point, thereby eliminating mechanical variables in the same batch of samples and improving the accuracy of drug action or gene editing effect assessment. This parameter system also provides an interface for subsequent closed-loop control, allowing real-time access to sensor feedback for online correction of the motion curve.

[0036] Reference Figure 1 and Figure 2 In some embodiments, the cell pulsation culture device includes a drive motor 600 and a lead screw. One end of the lead screw is connected to the drive motor 600, and the other end is connected to a second connecting plate 300. The drive motor 600 drives the lead screw to move according to preset parameters. By directly coupling the drive motor 600 and the lead screw, rotational motion is converted into linear displacement of the second connecting plate 300. This allows the waveform generation process to rely on only a single kinematic pair, simplifying the transmission chain and eliminating mechanical backlash caused by gear backlash, belt creep, or connecting rod clearance. The inherent helical self-locking characteristic of the lead screw can maintain the position of the second connecting plate 300 when the motor 600 is de-energized, preventing the rebound force of the elastic reaction chamber 400 from pushing the slide back, ensuring accurate stretching endpoints, and providing a reliable platform for experiments requiring phased steady-state observation.

[0037] Furthermore, the cell pulsation culture device includes a control unit, which is configured to control the drive motor 600 to operate according to preset parameters. Since the rotational speed of the motor 600 is linearly related to the lead screw, the control unit only needs to adjust the pulse frequency of the motor 600 to continuously change the moving speed, acceleration, and stroke of the second connecting plate 300. Corresponding to the preset parameters of the aforementioned embodiment, taking a specific embodiment as an example: the control unit adopts a discrete architecture of the programmable motor 600 control board and the motor 600 driver. First, the insulation at the ends of the signal wires and power wires is stripped with wire strippers. Then, the exposed conductors are pressed into tubular terminals with crimping pliers. Finally, the terminals are bolted to the corresponding terminals of the control board, driver, and 24V DC power supply: the direction signals D+ and D- correspond one-to-one with the pulse signals P+ and P-. The positive and negative terminals of the power supply are connected in parallel to the driver and motor 600 via a dedicated terminal block, forming a full-duplex, plug-and-play electrical link. After completing the wiring, the "Program Management" menu is accessed on the human-machine interface of the control board.

[0038] Specifically, a new file is created and specific relative motion, delay, and cycle commands are entered sequentially. By assigning displacement, velocity, and acceleration values ​​to each motion segment, typical pulse wave components generated by myocardial contraction, such as rapid rise, peripheral reflection wave, tidal plateau, pressure drop, isthmus descending, and dicrotic wave, can be written into the program in chronological order. After the command table is saved with one click, it is converted into a pulse sequence, driving the motor 600 to move forward and backward according to the step angle calculated by the lead screw, thereby enabling the second connecting plate 300 to reproduce a complete pulse waveform with physiological significance. If it is necessary to simulate hypertension, aortic regurgitation, or high discharge state after exercise, simply increase the acceleration, extend the plateau delay, or shorten the total cycle in the motion segment to instantly generate the corresponding pathological waveform.

[0039] Below is an example of the process for writing a loop instruction with a period of 1 second and constant acceleration: "Program Management" --- "Program Editing" --- Press and hold the "F" key to create a new file. Use the "Instruction Up" or "Instruction Down" keys to find "Relative Motion". Enter 1 for the label, 200 for the X-axis, and 500 for the velocity F. Press "Next Page" after inputting the information; use the "Instruction Down" keys to find the "Delay" instruction. Enter 2 for the label, and 250 for a delay of 0.25 seconds; find the "Relative Motion" instruction, enter 3 for the label, -200 for the X-axis, and 500 for F; find the "Delay" instruction. Enter 4 for the label, and 250 for a delay of 0.25 seconds; use the "Instruction Down" keys to find "Loop", enter 5 for the label, enter 9999 for the number of loops (representing an infinite loop), and enter 1 for the destination label to jump to the next loop. Complete the program writing, press "Exit" --- "Save Program", enter a filename, and click "OK" to save the file. If more complex frequencies or accelerations are required, the acceleration can be adjusted by searching for the "Specific Relative Motion" command, and the delay time can also be adjusted to achieve different force stimulation waveforms.

[0040] Blood does not flow steadily in blood vessels. Instead, with each heartbeat, the left ventricle ejects blood into the aorta, generating and propagating a pressure wave within the arterial system. The pulse wave contains a complex waveform with features including the ascending limb, main wave, tidal wave, descending isthmus, and dicrotic wave. Different physiological or pathological states correspond to drastically different pulse wave morphologies. Vascular endothelial cells and smooth muscle cells in the body sense this dynamic and complex mechanical environment. Therefore, using real pulse waves for stimulation is essential to obtain drug screening results that are more physiologically and pathologically relevant.

[0041] Furthermore, for the control unit, in some embodiments, the pulse wave simulation waveform programming process is exemplified as follows: 1) Locate the "Specific Relative Motion" command, enter 1 for the label, 80 for the X-axis, 1200 for the velocity F, and 5000 for the acceleration A (simulating the rapid rise of strong myocardial contraction). 2) Locate the "Delay" command, enter 2 as the label, and enter 50 (delay 0.05s, maintain high voltage state); 3) Locate the "Specific Relative Motion" command; input 3 for the label, 40 for the X-axis, 600 for the velocity F, and 3000 for the acceleration A (simulating the reflected wave of blood impacting peripheral blood vessels). 4) Locate the "Delay" command, enter 4 as the label, and enter 80 (delay 0.08s to maintain the tidal wave platform). 5) Find the "Specific Relative Motion" command label and enter 5, X-axis input -60, velocity F input 800, acceleration A input 4000 (simulate a sudden pressure drop); 6) Locate the "Delay" command, enter 6 as the label, and enter 30 (delay 0.03s, descend to the bottom of the gorge). 7) Locate the "Specific Relative Motion" command, enter 7 as the label, 25 as the X-axis, 400 as the velocity F, and 2000 as the acceleration A (simulating the elastic fluctuation after the aortic valve closes). 8) Locate the "Specific Relative Motion" command, enter 8 as the label, -85 as the X-axis, 500 as the velocity F, and 2500 as the acceleration A (to complete the diastolic descent phase). 9) Locate the "Delay" command, enter 9 as the label, and enter 680 (adjust the total cycle to 1 second, corresponding to a heart rate of 60 beats / minute). 10) Locate the "Loop" command, enter 10 as the label, enter 9999 for the number of loops, and enter 1 as the target label to establish an infinite loop simulating a continuous pulse. The device can be started or stopped according to the experimental usage time.

[0042] In summary, the above pulse wave simulation waveform programming process breaks down the complete heartbeat cycle into programmable micro-steps through specific relative motion, delay, and cycle instructions: First, a high-acceleration forward displacement simulates the rapid ascending limb caused by left ventricular ejection, followed by a brief delay to maintain the aortic peak; then, a slow forward advance reproduces the tidal wave plateau formed by peripheral vascular reflection waves; next, a high-acceleration reverse pull generates the descending isthmus with a sudden drop in pressure, and a small-amplitude forward displacement is used to superimpose the dicrotic wave after aortic valve closure; finally, a long delay completes the diastolic descending limb and returns to zero, forming a standard pulse; by connecting the ten segments of motion end to end through cycle instructions, a periodic waveform can be continuously output within the lead screw range. Since the displacement, velocity, and acceleration values ​​of each step correspond to the axial movement of the second connecting plate 300, researchers only need to enter the instructions in the order of the numbers on the control panel interface to make the drive motor 600 push the second connecting plate 300 according to the preset pulse sequence, thereby enabling the elastic reaction chamber 400 to reproduce the physiological-level pulsating stretching including the ascending branch, main wave, tidal wave, descending isthmus, and dicrotic wave. The normal or pathological state can be switched instantly without replacing mechanical parts.

[0043] Reference Figure 4 and Figure 7 In some embodiments, the reaction chamber 400 includes a main body 410 and two pins 420. The main body 410 defines a receiving chamber for receiving a target sample. Along the direction from the second connecting plate 300 to the first connecting plate 200, the two pins 420 are respectively inserted at both ends of the main body 410, and are respectively recessed into a first mounting groove 210 and a second mounting groove 310. By embedding rigid pins 420 at both ends of the elastic main body 410, the outer side of the pins 420 is in rigid contact with the mounting groove, ensuring that axial tensile force is transmitted to the entire chamber without slippage. The inner side of the pins 420 is fixed to the elastic main body 410 by covering, gluing, or snapping, allowing tensile stress to be evenly diffused to the sidewalls of the receiving chamber, avoiding localized stress concentration and membrane tearing caused by traditional clamping. This separable structure also provides flexibility in assembly and maintenance; researchers can replace the aged or contaminated main body 410 separately while retaining the metal pins 420, reducing long-term operating costs. Meanwhile, the engagement of the pin 420 with the shaft hole of the mounting slot allows the reaction chamber 400 to automatically center at the moment of clamping, ensuring that all samples are subjected to the same force direction without additional adjustment, thus reducing batch-to-batch differences.

[0044] Reference Figure 7 In some embodiments, the main body 410 includes a central region 411 and an edge region 412 surrounding the central region 411. The central region 411 is recessed to form a receiving chamber. Along the direction from the second connecting plate 300 to the first connecting plate 200, the size of the edge region 412 is larger than the size of the central region 411. Along the direction perpendicular to the direction from the second connecting plate 300 to the first connecting plate 200, the size of the edge region 412 is larger than the size of the central region 411. The recessed central region 411 forms a thin-walled receiving chamber, ensuring that axial tensile strain is mainly concentrated on the cell attachment surface. The thickened edge region 412 forms circumferential reinforcing ribs, which can suppress excessive lateral expansion of the sidewalls, making the mechanical transmission more uniform and reducing optical distortion. At the same time, the edge region 412 forms a flexible transition zone between the pin 420 and the central thin region, buffering clamping stress and avoiding early fatigue cracking of traditional equal-thickness films due to stress concentration, extending the service life of the reaction chamber 400 and improving the reliability of long-term pulsation experiments.

[0045] In some embodiments, the main body 410 includes a protrusion disposed within the receiving chamber to divide the receiving chamber into a first chamber and a second chamber that are interconnected along the direction from the second connecting plate 300 to the first connecting plate 200. Specifically, the protrusion is integrally formed within the receiving chamber. This protrusion does not completely isolate the receiving chamber; it allows for free exchange of liquid and cells through a communication opening on the top or sidewall, thus creating a local strain gradient within the same reaction chamber 400. For example, the area near the pin 420 experiences greater tension, while the strain near the root of the protrusion is relatively smaller, thereby simultaneously obtaining two mechanical microenvironments in a single physical sample. Researchers can seed the same or different types of cells in the first and second chambers respectively, comparing the effects of differential strain on cell phenotype, signaling pathways, and drug response under the same pulsation cycle, thereby increasing the density of experimental information. Furthermore, the protrusion also acts as an internal reinforcing rib, limiting excessive lateral expansion of the chamber sidewalls during repeated stretching, ensuring that axial stress is transmitted more evenly throughout the chamber, and reducing material fatigue caused by localized stress concentration. Meanwhile, the protrusions increase the contact area between the chamber and the culture medium, which is beneficial for gas exchange and nutrient diffusion, providing a more stable physicochemical microenvironment for long-term dynamic culture. This integrated partitioned design can achieve gradient mechanical stimulation without additional accessories, providing a simple, reliable, and economical solution for high-throughput drug screening and multi-parameter mechanism research.

[0046] Reference Figure 7 In some embodiments, the reaction chamber 400 is configured as a rectangular chamber with a wall thickness ranging from 4mm to 8mm (e.g., 4mm, 5mm, 6mm, 7mm, 8mm), a length ranging from 40mm to 60mm (e.g., 40mm, 50mm, 60mm), and a width ranging from 15mm to 30mm (e.g., 15mm, 20mm, 30mm). By defining the reaction chamber 400 as a rectangular cross-section and maintaining a moderate range in the wall thickness, length, and width, the elastomer possesses sufficient structural depth to withstand axial tension while retaining good light transmittance and space utilization. The rectangular outline and the planar sidewalls of the mounting groove ensure that the tensile force is evenly distributed across the entire end face. In some embodiments, the overall dimensions of the rectangular reaction chamber 400 are compatible with commonly used well plates and carriers, allowing direct placement into standard incubators or automated liquid workstations. This enables batch medium changes, staining, and imaging without additional fixtures, facilitating high-throughput, standardized cell mechanics experiments.

[0047] Reference Figures 8 to 10 The second aspect of this application provides a drug screening method, including the cell pulsating culture apparatus described above, with reference to... Figure 10 Drug screening methods include: S101: The same cells were seeded and different candidate substances were added in multiple reaction chambers 400; S103: Select the waveform parameters for simulating pathological pulsations and apply pulsation stretching; S105: Detects calcium deposition, apoptosis, or phenotypic transformation indicators in cells within 400 μm of each reaction chamber; S107: Compare the differences in indicators to screen for active substances that can antagonize the pathological pulsation effect.

[0048] Specifically, in some embodiments, S101 involves processing a batch of cells in parallel using multiple reaction chambers 400. First, an equal volume of cell suspension with the same genetic background and passage number is injected into each reaction chamber 400. Then, small molecules, antibodies, or herbal extracts to be screened are added accordingly, ensuring that drug differences are the only variable and eliminating systematic errors caused by inoculation time differences or concentration gradients in traditional multi-well plates. S103 involves uniformly applying pulsating waveforms simulating pathological features such as hypertension, arteriosclerosis, or valvular heart disease. The cells synchronously undergo dynamic stretching consistent with in vivo blood vessels, allowing potential active substances to exert their effects the instantaneously upon activation by the mechanically sensitive signal, avoiding the omission of force-drug coupling markers in static models. S105 utilizes calcium nodule staining, TUNEL apoptosis labeling, or immunofluorescence detection of osteogenic transcription factors to quantify changes in cell fate after mechanical stimulation into imageable and statistically significant objective indicators, achieving high-throughput parallel acquisition. By comparing the signal strength of the same indicator across different chambers, the S107 can intuitively determine which candidate substance can inhibit calcium deposition, reduce apoptosis, or block osteogenic phenotypic transformation, thereby precisely targeting active molecules with vascular protection, anti-fibrosis, or anti-calcification potential. The entire process is completed within a single device, eliminating the operational errors caused by the traditional method of stimulation, transfer, and then detection, providing a highly efficient and closed-loop approach for drug experiments on mechanically related diseases.

[0049] The following section describes the drug screening method in this application using the multi-channel pulsatile cell culture device for rapid screening of anti-calcification drugs as an example. Specifically, vascular calcification (VC) is a pathological phenomenon characterized by the abnormal deposition of calcium salts within the blood vessel wall, widely present in the progression of diseases such as arteriosclerotic vascular disease (ASVD), diabetes mellitus (DM), and chronic kidney disease (CKD). The osteogenic model of vascular smooth muscle cells is one of the important models for studying the mechanism of vascular calcification and screening potential therapeutic drugs. In this model, vascular smooth muscle cells undergo osteogenic differentiation under the influence of specific inducing factors (such as high phosphorus, β-glycerophosphate, and bone morphogenetic proteins). It is worth noting that, in addition to biochemical stimulation, biomechanical factors (such as periodic stress stimulation) are also considered important pathophysiological factors driving phenotypic transformation and calcification processes in VSMCs. Therefore, high-throughput in vitro simulation of the mechanical stress on the blood vessel wall is crucial for a deeper understanding of calcification mechanisms and rapid assessment of potential drug interventions.

[0050] During calcification, vascular smooth muscle cells (VSMCs) gradually lose their typical cell phenotype and transform into cells with osteogenic or chondrogenic characteristics. In osteogenic culture medium, VSMCs were subjected to pulsatile stimulation at a 10% amplitude for varying durations, followed by Alizarin Red staining. Alizarin Red is a commonly used staining method for detecting calcium salt deposits (such as calcium nodules), and its staining is primarily based on the reaction of Alizarin Red with calcium ions (Ca). 2+ The specific binding reaction of ) forms a complex, and the darker the gray in the figure, the more severe the cell calcification.

[0051] Alizarin red staining results showed that under 10% cyclic strain, the degree of cell calcification gradually increased over time. Calcification was not obvious in the first 2 hours, but after 4 hours, obvious calcified areas could be observed under the microscope. As time increased to 6-8 hours, calcification increased, covering more than 90% of the entire field of view after 8 hours. Figure 8 This indicates that 10% mechanical strain in osteogenic culture medium can effectively promote calcification of VSMCs within a short period (4-8 hours). In contrast, vascular smooth muscle cells require approximately 300 hours to achieve the same result under normal static culture conditions. Figure 8 The extent of calcification covering the entire field of vision.

[0052] Given that cyclic stimulation can rapidly induce calcification in VSMCs, this application utilized a multi-channel pulsatile cell culture device to test the inhibitory effects of various traditional Chinese medicine extracts on VSMC calcification. To quantify calcium-phosphate deposition, alizarin red staining was employed, which visually reflects the amount of calcium deposition through grayscale levels: lower staining intensity indicates a better inhibitory effect of the traditional Chinese medicine extracts on calcium deposition; conversely, higher staining intensity indicates a higher degree of calcification. Six of the most representative traditional Chinese medicine extracts were selected from the experiment, and their inhibitory effects on VSMC calcification were demonstrated using alizarin red staining images. (Alizarin Red staining) Figure 9 It can be clearly seen that samples treated with Dendrobium, Ophiopogon japonicus, and Angelica sinensis showed less calcium deposition, with the stained areas being nearly light-colored and almost no obvious gray (dark) stained areas, indicating that these three extracts had a strong inhibitory effect on calcium deposition. In contrast, samples treated with Gardenia jasminoides, Pueraria lobata, and Alisma plantago-aquatica showed higher calcium deposition, with the stained areas exhibiting a clear gray to dark gray variation, indicating that these extracts had a weaker inhibitory effect on calcification.

[0053] In summary, this application not only successfully constructed and validated a low-cost, high-throughput cellular stress application platform capable of highly simulating physiological pulsating stress environments, but also efficiently applied it to the rapid screening of anti-vascular calcification drugs, successfully identifying candidate drugs with inhibitory effects on calcification. The core value of this platform lies in its integration of dynamic biomechanical factors into the drug screening system, providing a powerful tool for more accurately simulating pathological processes closely related to the biomechanical microenvironment, such as vascular calcification. This not only expands the evaluation dimensions of vascular calcification intervention strategies, but more importantly, the platform provides an innovative high-throughput screening tool for simulating key pathological processes of the biomechanical microenvironment (such as vascular remodeling, osteoarthritis, and fibrosis), demonstrating broad application potential in the field of mechanobiology-guided drug development.

[0054] Reference Figure 11 The third aspect of this application provides a manufacturing process for fabricating the reaction chamber 400 of any of the above-described cell pulsation culture devices. The manufacturing process includes: S201: A cavity mold structure is formed by photopolymerization 3D printing, and the cavity mold structure corresponds to the outer surface of the reaction chamber 400; S203: The concave mold is irradiated with ultraviolet light for 2.5h~3.5h, and then placed in an oven at 65℃~75℃ for 4.5h~6.5h to eliminate the leached phosphine oxide photoinitiator in the structure; S205: Degassed liquid polydimethylsiloxane is poured into a concave mold and vacuum degassed; after curing at 65℃~75℃ for 1.5h~2.5h, it is demolded to obtain the elastic reaction chamber 400 body; S207: Insert pins 420 into both ends of the reaction chamber 400 and seal them to obtain the finished reaction chamber 400.

[0055] Specifically, in some embodiments, S201 uses photopolymer 3D printing to form a one-time mold, which allows for free design of the chamfers, reinforcing ribs, and draft angles of the reaction chamber's 400mm outer contour, eliminating the need for traditional machining or metal mold making, shortening the iteration cycle, and reducing small-batch costs. The digital files also ensure complete consistency in the morphology of different batches of molds, laying the geometric foundation for subsequent multi-channel parallel experiments. S203 uses a stepped treatment of UV followed by high temperature to completely decompose the residual phosphine oxide photoinitiator in the mold, preventing this substance from poisoning the platinum catalyst during subsequent PDMS casting, leading to incomplete curing or surface stickiness, thereby improving yield and biocompatibility. Simultaneously, heat treatment releases internal stress, preventing microcracks in the mold during the vacuum degassing stage. Specifically, the UV irradiation time for the mold can be 2.5h, 3h, or 3.5h, preferably 3h; the oven treatment temperature can be 65℃, 70℃, or 75℃, preferably 70℃; and the oven treatment time can be 1.5h, 2h, or 2.5h, preferably 2h. S205 involves vacuum degassing of liquid polydimethylsiloxane followed by isothermal curing, which completely eliminates bubble defects, ensuring uniform light transmittance and wall thickness of the elastic reaction chamber 400. This guarantees consistency in optical path and mechanical properties during subsequent pulsed stretching. The mild curing temperature protects the precision of the die and allows the PDMS cross-linked network to fully form, giving the material long-term fatigue resistance. S207 involves embedding prefabricated rigid pins 420 into both ends of the cured elastic body and forming a seamless transition through silicone adhesive or secondary curing. This utilizes the hard fit between the pins 420 and the mounting groove for reliable positioning while retaining the body's high elastic deformation capacity in the axial direction. This allows the finished reaction chamber 400 to be directly installed into a cell pulsed culture device for multiple assembly and disassembly without damaging the sealing surface, providing a durable and standardized culture chamber for high-throughput, reproducible drug screening experiments.

[0056] Furthermore, after step S107, the following steps are also included: The reaction chamber 400 body was cleaned with anhydrous ethanol, followed by ultrasonic cleaning for 15-25 minutes to remove surface residues. After cleaning, the reaction chamber 400 is immersed in 70%~75% alcohol for 0.8h~1.2h, then air-dried and sterilized by ultraviolet light for 2.5h~3.5h. After rinsing with sterile PBS and air-drying again, the reaction chamber was pre-washed with cell culture medium to complete sterilization and pre-equilibration of the reaction chamber 400.

[0057] Specifically, the step of "cleaning the reaction chamber 400 body with anhydrous ethanol, followed by ultrasonic cleaning for 15-25 minutes to remove surface residues" involves first rinsing with anhydrous ethanol and then using ultrasonic cavitation to rapidly dissolve the mold release agent, PDMS low-molecular-weight oligomers, and surface-adsorbed organic impurities, preventing these residues from altering the surface energy of the chamber wall and affecting cell adhesion. The subsequent alcohol immersion utilizes its dual effects of lipid solubility and protein denaturation to further extract deep-seated leachates and achieve preliminary sterilization, creating a low-bioload environment for subsequent sterilization. The ultrasonic cleaning time can be 15 minutes, 20 minutes, or 25 minutes, with 20 minutes being preferred.

[0058] The step "immersing the cleaned reaction chamber 400 in 70%~75% alcohol for 0.8h~1.2h, then air-drying and sterilizing under ultraviolet light for 2.5h~3.5h" employs a combined sterilization method of air drying and ultraviolet irradiation: air drying allows the alcohol to evaporate without leaving chemical residues that inhibit cell growth, while deep ultraviolet light destroys the nucleic acids of microorganisms. Compared to high temperature and high pressure, this method is more suitable for treating thermosetting elastomers, achieving deep sterilization without causing material deformation. The alcohol concentration can be 70%, 73%, or 75%, the immersion time can be 0.8h, 1h, or 1.2h, and the ultraviolet irradiation sterilization time can be 2.5h, 3h, or 3.5h.

[0059] The step "rinsing with sterile PBS and air-drying again, followed by pre-rinsing with cell culture medium to complete the sterilization and pre-equilibration of the reaction chamber 400" involves rinsing with sterile PBS to neutralize ozone generated by ultraviolet light and remove dead bacterial fragments. After a short air-drying process, residual ions are removed. Finally, pre-rinsing with complete culture medium allows the chamber walls to adsorb serum proteins and amino acids, forming a microenvironment consistent with the subsequent cell culture medium. This avoids osmotic pressure fluctuations caused by the instantaneous adsorption of nutrients from the culture medium on the PDMS surface, and ensures that cells are immediately in a familiar matrix interface after inoculation, improving survival rate and spreading uniformity. This, in turn, guarantees the biological reproducibility of the pulsatile stretching experiment and the reliability of drug screening data.

[0060] Below, refer to Figures 1 to 11This application systematically illustrates the advantages of this application through a specific embodiment. This application systematically integrates physiological-grade pulsatile stretching and high-throughput drug evaluation onto the same platform. The cell pulsatile culture device uses a base 100 as its framework, with a first connecting plate 200 and a second connecting plate 300 positioned parallel to each other. Pairs of mounting slots are arrayed along the vertical direction on the plate surfaces. Rigid pins 420 are pre-embedded at both ends of the elastic reaction chamber 400, allowing for rapid insertion into the corresponding slots for axial positioning. Quick clamping is achieved through pre-fabricated crossbars at both ends. Driven by a closed-loop stepper motor 600 and a precision lead screw, the second connecting plate 300 reciprocates according to preset speed, acceleration, and displacement parameters, thereby applying programmable stretching within the 0–200% engineering strain range to the reaction chamber 400. The motor 600 control board defines displacement, speed, acceleration, and delay segment by segment through an instruction table, thus reproducing a complete pulse wave including the ascending branch, main wave, tidal wave, descending isthmus, and dicrotic wave, providing a dynamic mechanical microenvironment completely consistent with in vivo conditions for vascular endothelial cells or smooth muscle cells. The entire mechanical system is manufactured using laboratory-grade photopolymerization and fused deposition modeling (FDM) 3D printing, eliminating the need for precision metal machining and reducing hardware costs. Simultaneously, the array-type mounting slots can be expanded to accommodate up to 96 independent reaction chambers 400, significantly increasing throughput compared to traditional 16-channel devices. Each chamber is a disposable PDMS chamber, completely eliminating cross-contamination. The modular printing platform allows for interchangeable 18, 48, or 96-channel specifications, accommodating rectangular reaction chambers 400 of varying lengths and widths to meet a range of needs from small-scale mechanistic studies to large-scale drug screening. Chamber wall thickness, pin dimensions (420mm), and mounting slot tolerances are precisely defined in the printing file, ensuring high repeatability and maintaining amplitude consistency across multiple chambers, reducing systematic errors. The rapid clamping and unclamping design allows for quick transfer of the reaction chambers 400 between incubators, laminar flow hoods, and microscopes, enabling continuous observation of real-time cell responses to pulsatile stimulation. Using this biomimetic mechanical model, researchers were able to observe calcium nodule deposition in vascular smooth muscle cells within hours under osteogenic induction conditions. Subsequently, they compared the inhibitory effects of different traditional Chinese medicine extracts on calcium deposition using Alizarin Red staining, rapidly identifying the effective active substances. In summary, this approach, centered on low-cost 3D printing and combined with high-precision motor 600 control and physiological waveform reproduction, provides an efficient, reliable, and easily scalable experimental platform for high-throughput drug screening of mechanically related diseases.

[0061] The above are merely preferred embodiments of the present invention and do not limit the scope of the patent. Any equivalent structural transformations made using the contents of the specification and drawings of the present invention under the inventive concept of the present invention, or direct / indirect applications in other related technical fields, are included within the scope of patent protection of the present invention.

Claims

1. A cell pulsation culture device, characterized by, The cell pulsation culture device comprises: a base, a first connecting plate connected to the base, the first connecting plate being provided with a plurality of first mounting slots arranged at intervals in a vertical direction, a second connecting plate connected to the base in a sliding manner, the second connecting plate being configured to be capable of moving closer to or away from the first connecting plate in a horizontal direction, the second connecting plate being provided with a plurality of second mounting slots corresponding to the first mounting slots in a vertical direction, a reaction chamber having two ends arranged oppositely in a horizontal direction, one end of the reaction chamber being arranged in the first mounting slot, the other end of the reaction chamber being arranged in the second mounting slot, the reaction chamber being used for accommodating a target sample, wherein the reaction chamber is elastic, and the second connecting plate is configured to be capable of moving closer to or away from the first connecting plate periodically according to a preset parameter, so that the reaction chamber provides at least one waveform of ascending branch, main wave, tidal wave, descending branch, and dicrotic wave to the target sample.

2. The cell pulsation culture device according to claim 1, wherein The cell pulsation culture device further comprises a third connecting plate connected to the base, the third connecting plate being located on a side of the second connecting plate away from the first connecting plate, the third connecting plate being provided with a plurality of third mounting slots arranged at intervals in a vertical direction, and a side of the second connecting plate facing the third connecting plate being provided with a plurality of fourth mounting slots corresponding to the third mounting slots, the third mounting slots and the fourth mounting slots being cooperated to mount the reaction chamber. The second connecting plate is configured to be capable of adjusting distances to the first connecting plate and the third connecting plate simultaneously in a horizontal direction.

3. The cell pulsation culture device according to claim 1, wherein The preset parameter at least includes an average speed of movement of the second connecting plate, an acceleration of movement of the second connecting plate, and a distance of movement of the second connecting plate.

4. The cell pulsation culture device according to claim 4, wherein The cell pulsation culture device comprises a driving motor and a lead screw, one end of the lead screw being connected to the driving motor, the other end of the lead screw being connected to the second connecting plate, and the driving motor driving the lead screw to move according to the preset parameter. And / or, The cell pulsation culture device comprises a control unit configured to control the driving motor to act according to the preset parameter.

5. The cell pulsation culture device according to claim 1, wherein The reaction chamber comprises a main body and two latches, the main body defining an accommodation chamber used for accommodating the target sample, and the two latches being respectively arranged at two ends of the main body in a direction from the second connecting plate to the first connecting plate, and the two latches being respectively arranged in the first mounting slot and the second mounting slot.

6. The cell pulsation culture device according to claim 5, wherein The main body comprises a central region and an edge region surrounding the central region, the central region being recessed to form the accommodation chamber, the edge region being larger in size than the central region in a direction from the second connecting plate to the first connecting plate, and the edge region being larger in size than the central region in a direction perpendicular to the direction from the second connecting plate to the first connecting plate.

7. The cell pulsation culture device according to claim 5, wherein The reaction chamber has a wall thickness ranging from 4 mm to 8 mm, a length ranging from 40 mm to 60 mm, and a width ranging from 15 mm to 30 mm.

8. A method of drug screening, characterized by, The cell pulsation culture device of any one of claims 1-8, wherein the drug screening method comprises: S101: inoculating the same cells in multiple reaction chambers and adding different candidate substances; S103: selecting a waveform parameter simulating pathological pulsation and applying pulsation stretching; S105: detecting calcium deposition, apoptosis, or phenotype transformation indicators of the cells in each reaction chamber; S107: comparing the differences in the indicators to screen active substances capable of antagonizing the effects of pathological pulsation.

9. A manufacturing process characterized by, The reaction chamber for manufacturing the cell pulsation culture device of any one of claims 1-7, wherein the manufacturing process comprises: S201: forming a concave mold structure corresponding to the outer surface of the reaction chamber by light-cured 3D printing; S203: treating the concave mold with ultraviolet light for 2.5-3.5 hours, and then placing it in an oven at 65-75°C for 4.5-6.5 hours to eliminate the leached phosphine oxide-based photoinitiator in the structure; S205: pouring degassed liquid polydimethylsiloxane into the concave mold and vacuuming to remove bubbles; after curing at 65-75°C for 1.5-2.5 hours, demolding to obtain an elastic reaction chamber body; S207: embedding a plug at both ends of the reaction chamber and sealing to obtain a finished reaction chamber.

10. The fabrication process of claim 9, wherein, After the step S207, the following steps are further included: cleaning the reaction chamber body with anhydrous ethanol, followed by ultrasonic cleaning for 15-25 minutes to remove surface residues; immersing the cleaned reaction chamber in 70-75% alcohol for 0.8-1.2 hours, then air-drying and sterilizing under ultraviolet light for 2.5-3.5 hours; rinsing with sterile PBS and air-drying again, then pre-washing with cell culture medium to complete sterilization and pre-equilibration of the reaction chamber.

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