Cell tissue culture device special for primary cells

Abstract

The invention relates to a cell tissue culture device special for primary cells, and relates to the technical field of cell culture, the cell tissue culture device comprises an incubator, the interior of the incubator is divided into three independent culture chambers through partition plates, each culture chamber is internally provided with at least three culture dishes, and the cell tissue culture device further comprises a dynamic culture assembly, at least two culture rooms are provided with a dynamic culture assembly, the dynamic culture assembly is provided with a micro-channel structure capable of simulating blood vessels, the dynamic culture assembly, the electrophysiological stimulation piece and the traction force simulation piece are integrated to form the culture system capable of simulating various physiological conditions in the body, the dynamic culture assembly simulates a blood vessel microchannel to realize low-shear-force continuous perfusion; the electrophysiological stimulation piece simulates a neuroelectric signal; the traction force simulation part applies controllable mechanical traction force through a multi-caliber breather pipe, and the three parts have a synergistic effect, so that the physiological function expression and culture quality of the primary cells are improved.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and in particular to a cell and tissue culture device specifically designed for primary cells. Background Technology

[0002] Primary cell culture is a core technology in modern life sciences, basic medical research, drug screening, and personalized clinical treatment. Its success highly depends on the ability to efficiently and non-destructively isolate highly active target cells from tissues. Tissue block culture, due to its advantages such as ease of operation, minimal cell damage, and better preservation of cell-cell interactions and the original in vivo microenvironment, has become the standard culture method for many important primary cells and is widely used. This method is commonly applied to the isolation and culture of fibroblasts, umbilical cord-derived mesenchymal stem cells, various epithelial cells, primary tumor cells, and muscle and nerve cells from human and animal tissues.

[0003] Traditional static culture methods have significant limitations. They not only easily lead to the accumulation of metabolic waste and uneven nutrient gradients, but also cannot simulate the complex physiological microenvironment in which cells exist in vivo. Many tissue cells (such as vascular endothelial cells and chondrocytes) need to withstand mechanical stimuli such as blood flow shear force, pressure, and tension in vivo. These stimuli are crucial for cell function expression and morphological maintenance. Static culture cannot meet these requirements, which restricts the quality and application value of primary cell culture. Based on this, a cell and tissue culture device specifically designed for primary cells is proposed. Summary of the Invention

[0004] To overcome the problems existing in related technologies, this invention provides a cell and tissue culture device specifically for primary cells. The dynamic culture component simulates vascular microchannels to achieve continuous perfusion with low shear force; the electrophysiological stimulation component simulates nerve electrical signals; and the tension simulation component applies controllable mechanical tension through multi-diameter ventilation tubes. The three components work synergistically to improve the physiological function expression and culture quality of primary cells.

[0005] To achieve the above objectives, the present invention provides a cell and tissue culture device specifically for primary cells, comprising an incubator divided into three independent culture chambers by a partition, each culture chamber containing at least three culture dishes, each culture dish being rotatable to the front opening of the incubator via a deflection assembly, and further comprising: A dynamic culture assembly is provided in at least two of the culture chambers. The dynamic culture assembly has a microchannel structure that can simulate blood vessels, enabling continuous, low-shear perfusion of the culture medium. A flexible membrane is disposed at the bottom of the interior of the culture dish; An electrophysiological stimulation device is disposed below the flexible membrane and is used to apply electrical pulses to simulate nerve signals; A tension simulation element is disposed below the flexible membrane, and the tension simulation element has multiple venting tubes with different inner diameters. The outlet end of the venting tube is sealed to the lower end face of the flexible membrane. The tension simulation element applies controllable mechanical tension to the primary cells.

[0006] Preferably, the dynamic culture component includes: The first main delivery pipe has a peristaltic pump installed at one end; A ring-shaped conduit passes through multiple culture dishes within the same culture chamber and is connected to the other end of the first main delivery tube; Multiple perfusion microtubes are located inside each culture dish and connected to a circular conduit, with each perfusion microtube having a different outlet direction.

[0007] Preferably, an air inlet pipe is connected to the first conveying main pipe, and a miniature valve is installed on both the first conveying main pipe and the air inlet pipe.

[0008] Preferably, the electrophysiological stimulation device includes: A tray is connected below the flexible membrane, and a groove is provided between the tray and the flexible membrane; A miniature stress sensor is installed on the lower end face of the flexible membrane; A miniature multi-electrode array is mounted in a ring array on the lower end face of the flexible membrane and arranged around the miniature stress sensor.

[0009] Preferably, the tension simulation element includes: trachea; The first annular tube and the second annular tube are concentrically arranged in the groove. The air tube connects the first annular tube and the second annular tube. Multiple first conduits are provided between the first annular tube and the flexible membrane, and multiple second conduits are provided between the second annular tube and the flexible membrane.

[0010] Preferably, the inner diameters of the plurality of first catheters are distributed in alternating large and small sizes, and the inner diameters of the plurality of second catheters are also distributed in alternating large and small sizes, with the inner diameter of the first catheters being larger than the inner diameter of the second catheters.

[0011] Preferably, the system also includes a sensor array, which is installed on the lower end of the side wall of the culture dish, and the sensor array includes, but is not limited to, a dissolved oxygen sensor, an impedance sensor, and a temperature sensor.

[0012] Preferably, the deflection component includes: A servo motor with a connecting rod that penetrates a partition at its top, and multiple sets of connecting strips fixed to the outside of the connecting rod; Multiple annular sliding plates, each of which is fixed below a set of connecting bars; An annular groove is formed on the upper end face of the partition plate, and the annular groove slides into a corresponding annular groove.

[0013] Preferably, it also includes a culture medium base, and the bottom of the culture dish in the culture chamber is provided with a culture medium base, which includes a first culture medium, a second culture medium, a third culture medium and a fourth culture medium with continuously varying stiffness.

[0014] Preferably, the dynamic culture component includes a second delivery main pipe that extends through a single culture dish and is connected to a plurality of transverse and longitudinal perfusion tubes.

[0015] The technical solution provided by this invention may include the following beneficial effects: 1. In this invention, a culture system that can simulate various physiological conditions in vivo is formed by integrating a dynamic culture component, an electrophysiological stimulation component, and a tension simulation component. The dynamic culture component simulates vascular microchannels to achieve continuous perfusion with low shear force; the electrophysiological stimulation component simulates nerve electrical signals; and the tension simulation component applies controllable mechanical tension through multi-diameter ventilation tubes. The three components work synergistically to effectively restore the mechanical and electrophysiological environment experienced by cells in vivo, thereby improving the physiological function expression and culture quality of primary cells.

[0016] 2. In this invention, the gradient stiffness culture substrate can simulate fibrosis or tissue softening environments. Combined with dynamic perfusion and mechanical stimulation, it can study cell behavior in different microenvironments in parallel. The multi-culture chamber and multi-culture dish design supports the co-culture of multiple cell types, which can simulate the material exchange and functional synergy between organs in vivo, providing a more reliable model for drug toxicity detection, tissue engineering and other research.

[0017] 3. In this invention, multiple culture dishes can be connected by microchannels of the dynamic culture component, which can construct a simplified culture model that simulates the synergistic effect of multiple organs, such as a tandem culture system of hepatocytes, kidney cells and vascular endothelial cells, providing an experimental platform that is closer to physiological reality for cell and tissue culture research.

[0018] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and are not intended to limit the invention. Attached Figure Description

[0019] The above and other objects, features and advantages of the present invention will become more apparent from the more detailed description of exemplary embodiments of the invention in conjunction with the accompanying drawings, wherein the same reference numerals generally represent the same parts in the exemplary embodiments of the invention.

[0020] Figure 1 This is a schematic diagram of the structure of the present invention; Figure 2This is a schematic diagram of the external structure of the present invention; Figure 3 This is a schematic diagram of the structure of the petri dish, deflection component, and dynamic culture component according to the first embodiment of the present invention; Figure 4 This is a schematic diagram of the deflection component of the present invention in its disassembled state; Figure 5 This is a schematic diagram of the structure of the dynamic cultivation component of the present invention in the first embodiment; Figure 6 This is a schematic diagram of the structure of the electrophysiological stimulation device, flexible membrane, and tension simulation device of the present invention; Figure 7 This is a schematic diagram of the electrophysiological stimulation device, flexible membrane, and tension simulation device of the present invention in their disassembled state; Figure 8 This is a cross-sectional structural schematic diagram of the electrophysiological stimulation device of the present invention; Figure 9 This is a schematic diagram of the structure of the culture medium substrate under the second embodiment of the dynamic culture component of the present invention; Figure 10 This is a cross-sectional structural diagram of the culture substrate under the second embodiment of the dynamic culture component of the present invention.

[0021] The correspondence between the labels and component names in the attached figures is as follows: 1. Incubator; 11. Door; 12. Unit door; 2. Culture chamber; 3. Petri dish; 4. Deflection assembly; 41. Servo motor; 42. Connecting rod; 43. Connecting bar; 44. Circular sliding plate; 45. Circular slide groove; 5. Dynamic culture assembly; 51. First main delivery tube; 52. Circular catheter; 53. Perfusion microtube; 54. Air inlet tube; 55. Second main delivery tube; 56. Horizontal perfusion tube; 57. Vertical perfusion tube; 6. Sensor array; 7. Electrophysiological stimulation device; 71. Support plate; 72. Groove; 73. Miniature stress sensor; 74. Miniature multi-electrode array; 8. Flexible membrane; 9. Tension simulation component; 91. Trachea; 92. First annular tube; 93. Second annular tube; 94. First catheter; 95. Second catheter; 10. Culture base; 101. First culture substrate; 102. Second culture substrate; 103. Third culture substrate; 104. Fourth culture substrate. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of the embodiments of this 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 this invention, and not all of them. All other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this invention. Preferred embodiments of the invention will now be described in more detail with reference to the accompanying drawings. Although preferred embodiments of the invention are shown in the drawings, it should be understood that the invention can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to make the invention more thorough and complete, and to fully convey the scope of the invention to those skilled in the art.

[0023] The technical solutions of the embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0024] Example 1: See Figures 1-8As shown, this invention proposes a cell and tissue culture device specifically for primary cells, including an incubator 1. A controller is mounted on the front of the outer wall of the incubator 1. This controller is equipped with a high-definition LCD screen and multiple touch buttons, which can not only display key parameters such as temperature and culture time in real time, but also allow operators to precisely set temperature thresholds according to the culture needs of different types of primary cells. It also has an alarm function for abnormal parameters; when the environment inside the incubator 1 deviates from the set range, it will promptly alert the operator through an audible and visual alarm. The incubator 1 is equipped with an electric heating element, and is divided into three independent culture chambers 2 by partitions. A door 11 is located on the front of the incubator 1, with independent unit doors 12 corresponding to each culture chamber 2. A transparent observation window can be installed on each unit door 12. Each culture chamber 2 contains at least three culture dishes 3, and each culture dish 3 can be rotated to the front opening of the incubator 1 via a deflection component 4. The device also includes a dynamic culture component 5, an electrophysiological stimulation component 7, a flexible membrane 8, and a tension simulation component 9. At least two culture chambers 2... The culture dish 3 is equipped with a dynamic culture component 5, which has a microchannel structure that can simulate blood vessels, enabling continuous, low-shear perfusion of the culture medium. A flexible membrane 8 is located at the bottom of the culture dish 3. The flexible membrane 8 is preferably made of PDMS due to its good biocompatibility and transparency. An electrophysiological stimulation device 7 is located below the flexible membrane 8 and is used to apply electrical pulses to simulate nerve signals. A tension simulation device 9 is located below the flexible membrane 8 and has multiple venting tubes with different inner diameters. The venting tubes are sealed to the lower end of the flexible membrane 8. The tension simulation device 9 applies controllable mechanical tension to the primary cells. A sensor group 6 is installed inside the culture dish 3. The sensor group 6 is installed at the lower end of the side wall of the culture dish 3 and includes, but is not limited to, a dissolved oxygen sensor, an impedance sensor, and a temperature sensor. The dissolved oxygen sensor monitors the metabolic state in real time. The impedance sensor can monitor cell adhesion, proliferation, and morphological changes (such as cell membrane integrity) in real time without labeling. The temperature sensor is used to monitor the temperature and can be used with a controller and an electric heating key to control the culture temperature.

[0025] Among them, see Figures 3-4As shown, the deflection assembly 4 includes a servo motor 41 and multiple annular slide plates 44. The servo motor 41 is installed at the bottom of the incubator 1. A connecting rod 42 that penetrates the partition is installed at the top of the servo motor 41. A bearing is installed at the position where the connecting rod 42 penetrates the partition to achieve stable rotation of the connecting rod 42. Multiple sets of connecting strips 43 are fixed to the outside of the connecting rod 42. Each annular slide plate 44 is fixed below a set of connecting strips 43. The culture dish 3 is placed on the upper surface of the annular slide plate 44. To increase friction, anti-slip textures can be provided on the upper surface of the annular slide plate 44 and the bottom of the culture dish 3 to effectively prevent the culture dish 3 from sliding or shifting during the rotation of the annular slide plate 44, ensuring the culture dish 3 is placed safely. The partition is securely positioned, and an annular groove 45 is provided on the upper surface of the partition. The annular groove 45 slides and engages with the corresponding annular groove 45. When the servo motor 41 is started, the output end of the servo motor 41 drives the connecting rod 42 to rotate. This drives the annular slide plate 44 to rotate along the annular groove 45 through the connecting bar 43, so that the culture dish 3 to be taken out is rotated to the front opening of the incubator 1 for easy removal by the experimenter. This significantly shortens the operation distance when the experimenter takes out the culture dish 3, which not only improves the efficiency of taking out and placing the culture dish 3, but also reduces the extent to which the arm needs to penetrate into the incubator 1, reduces the interference with the stable environment inside the incubator 1, and makes it easier for the experimenter to quickly and safely complete the operation of taking out and placing the culture dish 3.

[0026] See Figure 1 as well as Figures 3-5As shown, the dynamic culture assembly 5 includes a first delivery main tube 51, a circular conduit 52, and multiple perfusion microtubes 53. The first delivery main tube 51 is a flexible tube of sufficient length. One end of the first delivery main tube 51 is used to install a peristaltic pump, which needs to be equipped with a pulse damper to ensure stable flow rate and avoid excessive pulse shear force from damaging the cells. The circular conduit 52 runs through multiple culture dishes 3 within the same culture chamber 2. A micro-valve, which is an electrically controlled valve, is installed on the circular conduit 52. A porous membrane is provided inside the circular conduit 52 and at the ends of each culture dish 3 and on the perfusion microtubes 53. Nano- to micron-sized pores are created on the membrane to prevent the backflow of cells and macromolecular metabolic waste from the culture dishes 3 into the conduit, while allowing small molecules of culture medium (such as glucose, oxygen, and growth factors) in the circular conduit 52 to diffuse freely to the conduit. Wall cells are connected to the other end of the first delivery main pipe 51. Perfusion microtubes 53 are located inside each culture dish 3 and connected to the annular conduit 52. Each perfusion microtube 53 has a different outlet direction. The culture medium is dynamically delivered via a peristaltic pump. The culture medium can enter the annular conduit 52 through the first delivery main pipe 51 and then slowly flow out through the perfusion microtubes 53 in different directions. This multi-directional perfusion method not only ensures that fresh culture medium evenly covers all cells in the culture dish 3, avoiding local nutrient deficiency, but also creates a gentle and continuous fluid shear force through liquid flow. This shear force simulates the physiological stimulation of cells by blood flow in vivo. This stimulation can regulate cell morphology, polarity, and functional expression, helping to cultivate cells whose function is closer to the real state in vivo, far exceeding the limitations of single-function cells under static culture. To change the functional limitations of a single culture chamber, a chamber structure containing multiple culture dishes 3 is designed. Each culture dish 3 is interconnected through a microchannel structure, constructing a simplified culture model simulating the synergistic effect of multiple human organs.

[0027] Taking a culture chamber 2 as an example, the three culture dishes 3 inside are used to culture primary hepatocytes, primary kidney cells, and vascular endothelial cells, respectively. These three types of cells are key cell types in the human metabolic and circulatory systems. Primary hepatocytes are responsible for substance metabolism and detoxification, primary kidney cells are responsible for filtration and excretion, and vascular endothelial cells are the core cells that make up the blood vessel wall. The synergistic effect of the three is an important link in the human circulatory metabolism. The microchannel structure connecting each culture dish 3 not only undertakes the task of transporting culture medium, but also simulates the human vascular network, connecting the culture dishes 3 containing the three types of cells into a closed-loop system.

[0028] During the culture process, as the culture medium carrying nutrients flows through the vascular endothelial cell culture dish 3, the endothelial cells secrete vasoactive substances that dissolve into the culture medium. Subsequently, the culture medium continues to flow to the hepatocyte culture dish 3, providing nutrients to the hepatocytes. The intermediate products of hepatocyte metabolism and the detoxified substances then enter the kidney cell culture dish 3 with the culture medium, where the kidney cells complete the filtration and metabolism of related substances. This design not only replicates the material exchange and functional synergy between cells of different organs, but also simulates the paracrine regulatory mechanism between organs in the body, allowing cells to grow under conditions closer to the physiological environment, thus improving the realism of cell culture and providing a more reliable cell model for subsequent studies such as drug toxicity testing and organ function simulation.

[0029] In addition, an air inlet pipe 54 is connected to the first delivery main pipe 51. A miniature air pump is installed at the end of the air inlet pipe 54. Both the first delivery main pipe 51 and the air inlet pipe 54 are equipped with miniature valves, which are electrically controlled valves. These valves can not only deliver culture medium to simulate blood vessel pressure, but also introduce treated air through the air inlet pipe 54. By utilizing the power output of the miniature air pump and the coordinated operation of the electrically controlled valves, positive pressure is applied periodically according to a preset program, thereby simulating the pressure changes in the thoracic cavity during human respiration. This satisfies the needs of cells or tissues for nutrient supply and metabolic environment, and can more comprehensively reproduce the complex physiological microenvironment in the body through the dynamic simulation of respiratory pressure, providing a more realistic model basis for related cell and tissue culture experimental research.

[0030] See Figures 6-8 As shown, the electrophysiological stimulation device 7 includes a support plate 71, a micro-stress sensor 73, and a micro-multi-electrode array 74. The support plate 71 is connected to the lower part of the flexible membrane 8, and a groove 72 is provided between the support plate 71 and the flexible membrane 8. The micro-stress sensor 73 is installed on the lower end face of the flexible membrane 8 and can monitor the force exerted on the flexible membrane 8 by cells during growth and contraction in real time. The micro-multi-electrode array 74 is installed in a ring array on the lower end face of the flexible membrane 8 and is arranged around the micro-stress sensor 73. It can dynamically adjust the applied mechanical tension according to the measured cell contraction force. For example, when the spontaneous cell contraction force is detected to be weakened, the stimulation intensity is automatically increased.

[0031] The electrophysiological stimulation device 7 can simultaneously apply two key stimuli: on the one hand, it releases electrical pulses through the micro multi-electrode array 74 to simulate real neural electrical signals in vivo, guiding neurons to form normal synaptic connections and maintain signal transmission function; on the other hand, it works with the dynamic culture component 5 to generate mechanical tension, simulating the mechanical environment of the heart during diastole and systole or the mechanical microenvironment of neural tissue. The two stimuli work synergistically to construct a physiological microenvironment close to that in vivo for primary neurons and cardiomyocytes, effectively promoting the maturation and stability of cell function, and providing reliable technical support for related cell research, drug screening and tissue engineering applications.

[0032] See Figures 6-8 As shown, the tension simulation component 9 includes an air tube 91, a first annular tube 92, and a second annular tube 93. The first annular tube 92 and the second annular tube 93 are concentrically arranged in the groove 72, and the first annular tube 92 is located on the periphery of the second annular tube 93. The air tube 91 connects the first annular tube 92 and the second annular tube 93. The air inlet end of the air tube 91 is connected to a micro air pump to provide a stable airflow power source for the entire tension simulation. Multiple first conduits 94 are provided between the first annular tube 92 and the flexible membrane 8, and multiple second conduits 95 are provided between the second annular tube 93 and the flexible membrane 8. The first conduits 94 and the second conduits 95 are ventilation tubes. The inner diameters of the multiple first conduits 94 are distributed alternately in large and small sizes, and the inner diameters of the multiple second conduits 95 are also distributed alternately in large and small sizes. The inner diameter of the first conduit 94 is larger than the inner diameter of the second conduit 95. The difference in the inner diameter design can achieve differentiated tension effects in different areas of the flexible membrane 8.

[0033] Through the above, the device can achieve multi-dimensional physiological environment simulation during cell and tissue culture. On the one hand, the dynamic culture component 5 continuously provides blood shear force simulation to the cells, restoring the mechanical stimulation of blood flow to the cells. On the other hand, the tension simulation component 9 is used to simulate physiological tension actions such as breathing or heartbeat. In specific operation, the air pressure output of the micro air pump is precisely controlled by a preset program to apply positive and negative pressure in a programmed manner. When positive pressure is applied, the airflow is orderly introduced into the first annular tube 92 and the second annular tube 93 through the trachea 91. After the two annular tubes are filled with gas and a stable air pressure is formed, the airflow will rush into the corresponding first conduit 94 and the second conduit 95 respectively. Since the two types of conduits not only have different inner diameters, but the inner diameters of the same type of conduits also change alternately, the pressure intensity formed by the airflow in conduits with different inner diameters is different, which in turn generates different thrusts on different areas of the flexible membrane 8, causing uneven expansion of the flexible membrane 8 locally. When the negative pressure is applied, the trachea 91 releases the gas in the annular tube and duct in an orderly manner. The thrust on the flexible membrane 8 gradually disappears and gradually contracts and returns to its original position under its own elasticity. Through this periodic switching between positive and negative pressure, the flexible membrane 8 can continuously generate cyclic tension. The variation pattern of this tension matches the expansion and contraction of the thoracic tissue during respiration and the rhythmic expansion and contraction of the myocardial tissue during heartbeat in the body. This creates a mechanical microenvironment that closely resembles the real state in the body for the cultured cells and tissues, thereby enhancing the activity and physiological stability of the cells.

[0034] Example 2: See Figure 1 as well as Figures 9-10As shown, the cell and tissue culture apparatus also includes a culture substrate 10. In addition, the culture substrate 10 is set at the bottom of the culture dish 3 inside the culture chamber 2. The culture substrate 10 includes a first culture medium 101, a second culture medium 102, a third culture medium 103, and a fourth culture medium 104 with continuously varying stiffness. It can be used to study how mechanical force and perfusion affect cell behavior (such as the migration of tumor cells) in fibrotic (increased stiffness) or tissue softening environments. The implementation of the stiffness gradient culture substrate 10 can be explained by the following steps: 1. Inject two liquids, A and B, at a constant flow rate using a precision injection pump; 2. The two liquids are mixed by laminar diffusion and convection, forming a crosslinking agent concentration gradient from one side (side A) to the other side (side B) when they enter the culture dish 3; 3. Fill the entire culture chamber with the mixed liquid; 4. Instantly trigger global curing. For photocurable hydrogels (such as GelMA and PEGDA), perform a single, uniform UV irradiation; for chemically cross-linked hydrogels (such as collagen), wait for them to undergo overall thermal gelation. 5. After curing, a hydrogel substrate with a stiffness that changes continuously from side A to side B is formed at the bottom of the culture chamber.

[0035] The dynamic culture component 5 includes a second delivery main pipe 55, which penetrates a single culture dish 3. Multiple transverse perfusion tubes 56 and longitudinal perfusion tubes 57 are connected to the second delivery main pipe 55. The inlet ends of the transverse perfusion tubes 56 and the longitudinal perfusion tubes 57 can be connected by flexible tubes to achieve synchronous perfusion.

[0036] As described above, since the culture medium substrate 10 has a gradient stiffness, when a uniform strain is applied, the local stress perceived by actual cells in different stiffness regions is different. Cells in high-stiffness regions experience greater tensile stress, while those in low-stiffness regions experience less. This allows for the simultaneous study of multiple mechanical microenvironments within the same culture chamber, eliminating the need for multiple independent culture devices. Multiple different mechanical microenvironments can be constructed simultaneously within the same culture chamber, facilitating parallel studies of cell physiological responses to different mechanical stimuli. In actual cell and tissue culture applications, the prepared culture medium solution can be delivered via the second delivery main pipe 55. After being diverted by the main pipe, these nutrient solutions are slowly and steadily injected into the culture dish 3 through interconnected transverse and longitudinal perfusion tubes 56 and 57. This avoids impacting the fragile cell structure while continuously providing fresh nutrients to the cells. Simultaneously, it simulates the dynamic circulation of bodily fluids such as blood and tissue fluid, further enhancing the complexity and realism of the physiological scenario simulation. This provides culture conditions that more closely resemble the actual in vivo state for studying cell behavior under different mechanical and nutritional environments.

[0037] The present invention has been described in detail above with reference to the accompanying drawings. In the above embodiments, the descriptions of each embodiment have their own emphasis; for parts not described in detail in a certain embodiment, please refer to the relevant descriptions of other embodiments. Those skilled in the art should also understand that the actions and modules involved in the specification are not necessarily essential to the present invention. Furthermore, it is understood that the steps in the method of the embodiments of the present invention can be adjusted, combined, and deleted according to actual needs, and the structure in the device of the embodiments of the present invention can be combined, divided, and deleted according to actual needs.

[0038] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or improvement of the technology in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A cell and tissue culture apparatus specifically for primary cells, comprising an incubator (1) divided into three independent culture chambers (2) by partitions, each of the culture chambers (2) containing at least three culture dishes (3), characterized in that, Each of the aforementioned petri dishes (3) is rotatable to the front opening of the incubator (1) via a deflection assembly (4), and further includes: Dynamic culture component (5), at least two of the culture chambers (2) are equipped with dynamic culture component (5), the dynamic culture component (5) has a microchannel structure that can simulate blood vessels, and can realize continuous, low shear force perfusion of culture medium solution; A flexible membrane (8) is disposed at the bottom of the interior of the culture dish (3); An electrophysiological stimulation device (7) is disposed below the flexible membrane (8) and is used to apply electrical pulses to simulate nerve signals; A tension simulation element (9) is located below the flexible membrane (8), and the tension simulation element (9) has multiple venting tubes with different inner diameters. The outlet end of the venting tube is sealed to the lower end face of the flexible membrane (8). A controllable mechanical tension is applied to the primary cells through the tension simulation element (9).

2. The cell and tissue culture apparatus specifically for primary cells according to claim 1, characterized in that, The dynamic culture component (5) includes: The first delivery main pipe (51) has one end for mounting a peristaltic pump; A ring-shaped conduit (52) passes through multiple culture dishes (3) within the same culture chamber (2) and is connected to the other end of the first delivery main pipe (51); Multiple perfusion microtubes (53) are located inside each culture dish (3) and connected to the ring conduit (52), and the outlet directions of each perfusion microtube (53) are different.

3. The cell and tissue culture apparatus specifically for primary cells according to claim 2, characterized in that, The first conveying main pipe (51) is connected to an air inlet pipe (54), and both the first conveying main pipe (51) and the air inlet pipe (54) are equipped with miniature valves.

4. The cell and tissue culture apparatus specifically for primary cells according to claim 1, characterized in that, The electrophysiological stimulation device (7) includes: A tray (71) is connected to the bottom of the flexible membrane (8), and a groove (72) is provided between the tray (71) and the flexible membrane (8). A miniature stress sensor (73) is mounted on the lower end face of the flexible membrane (8); A miniature multi-electrode array (74) is mounted in a ring array on the lower end face of the flexible membrane (8) and arranged around the miniature stress sensor (73).

5. The cell and tissue culture apparatus specifically for primary cells according to claim 4, characterized in that, The tension simulation component (9) includes: Trachea (91); The first annular tube (92) and the second annular tube (93) are concentrically arranged in the groove (72). The air tube (91) connects the first annular tube (92) and the second annular tube (93). A plurality of first conduits (94) are provided between the first annular tube (92) and the flexible membrane (8), and a plurality of second conduits (95) are provided between the second annular tube (93) and the flexible membrane (8).

6. The cell and tissue culture apparatus specifically for primary cells according to claim 5, characterized in that, The inner diameters of the multiple first catheters (94) are distributed in alternating large and small sizes, and the inner diameters of the multiple second catheters (95) are also distributed in alternating large and small sizes, and the inner diameter of the first catheter (94) is larger than the inner diameter of the second catheter (95).

7. The cell and tissue culture apparatus specifically for primary cells according to claim 1, characterized in that, It also includes a sensor group (6), which is installed on the lower side wall of the culture dish (3), and the sensor group (6) includes, but is not limited to, a dissolved oxygen sensor, an impedance sensor and a temperature sensor.

8. The cell and tissue culture apparatus specifically for primary cells according to claim 1, characterized in that, The deflection component (4) includes: A servo motor (41) has a connecting rod (42) that penetrates the partition plate installed at its top. Multiple sets of connecting strips (43) are fixed to the outside of the connecting rod (42). Multiple annular slide plates (44), each of which is fixed below a set of connecting strips (43); An annular groove (45) is formed on the upper end face of the partition plate, and the annular groove (45) is slidably fitted with the corresponding annular groove (45).

9. The cell and tissue culture apparatus specifically for primary cells according to claim 1, characterized in that, It also includes a culture substrate (10). In addition, a culture substrate (10) is provided at the bottom of the culture dish (3) inside the culture chamber (2). The culture substrate (10) includes a first culture substrate (101), a second culture substrate (102), a third culture substrate (103), and a fourth culture substrate (104) with continuously varying stiffness.

10. The cell and tissue culture apparatus for primary cells according to claim 8, characterized in that, The dynamic culture component (5) includes a second delivery tube (55) that extends through a single culture dish (3) and is connected to a plurality of transverse perfusion tubes (56) and longitudinal perfusion tubes (57).

Images