Cell bionic culture device, culture unit, and culture membrane

Abstract

An objective is to provide a cell bionic culture device, a culture unit, and a culture membrane. The cell bionic culture device comprises: a culture assembly comprising a plurality of culture units isolated from each other, each culture unit comprising a base and a culture membrane, wherein the base has a variable-pressure space connected to the culture membrane, the culture membrane flexibly deforms by means of pressure variations in the variable-pressure space, and the variable-pressure spaces of the plurality of culture units are independent of each other; and a fluid pressure supply assembly separately fluidly connected to the variable-pressure spaces independent of each other, so as to provide pressure variations in the variable-pressure spaces.

Description

Cell biomimetic culture equipment, culture unit, and culture membrane Technical Field

[0001] This application relates to cell biomimetic culture equipment, culture units, and culture membranes. Background Technology

[0002] Within living organisms, the growth, proliferation, death, migration, and differentiation of cells are closely related to their mechanical environment. Mesenchymal stem cells (MSCs) are widely available, possess strong proliferative capacity, and have the potential for multi-lineage differentiation. Mechanical stimuli (such as compressive, tensile, and fluid shear forces) play a crucial role in the normal growth and differentiation of MSCs. Mechanical stretching has been reported to be an important regulator of MSC behavior, promoting osteogenic differentiation and skin regeneration; therefore, research on cell mechanics has attracted increasing attention. To simulate the real growth environment of cells in vivo, external forces can be applied to stretch or compress cells during cell culture, resulting in biomimetic culture.

[0003] In some comparative schemes, the biomimetic culture component uses a vacuum system to stretch the flexible membrane on a porous culture plate. However, since each culture site on the porous culture plate is interconnected, it is difficult to individually control the stretching frequency and tension at each culture site.

[0004] In other comparative schemes, the culture membrane of the biomimetic culture component is fixed with screws / bolts to ensure stable fixation and uniform force distribution, but this makes it complicated to replace the culture membrane and makes it difficult to replace a single culture membrane individually.

[0005] Therefore, there is a need in the art for a new biomimetic cell culture device, culture unit, and culture membrane to solve one or a combination of the above problems. Summary of the Invention

[0006] The purpose of this application is to achieve individual control of the stretching frequency and tension of each culture position in a cell biomimetic culture device, so that a single culture can achieve comparative experiments under different conditions, thereby improving the efficiency of the culture device; and / or the culture membrane of the culture unit is easy to replace on the basis of stable fixation and uniform stress.

[0007] A cell biomimetic culture device according to a first aspect of this application includes: a culture assembly comprising a plurality of culture units isolated from each other; each culture unit comprising a base and a culture membrane; the base having a pressure-changing space connected to the culture membrane, wherein pressure changes in the pressure-changing space cause the culture membrane to undergo flexible deformation; wherein the pressure-changing spaces of the plurality of culture units are independent of each other; and a fluid pressure supply assembly fluidly connected to each of the independent pressure-changing spaces to provide pressure changes in the pressure-changing spaces.

[0008] In one or more embodiments of the culture device, the culture assembly further includes a mounting base having a plurality of mounting seats, each mounting seat corresponding to a single culture unit; preferably, the base has a fluid channel, and the pressure-changing space is in communication with the fluid channel; the fluid channel inlets of the respective fluid channels of the plurality of culture units are independent of each other.

[0009] In one or more embodiments of the culture device, the culture membrane includes a membrane body and a membrane connecting portion located around the membrane body. The base includes a mounting boss, the top surface of which is disposed corresponding to a portion of the membrane body. The pressure-changing space is located around the mounting boss and is disposed corresponding to another portion of the membrane body. The other portion of the membrane body is located around the portion of the membrane body. Preferably, the base includes a first connecting portion corresponding to the membrane connecting portion, and a radial gap between the first connecting portion and the mounting boss provides the pressure-changing space. More preferably, the base has a first wall, a second wall, and a third wall distributed sequentially from the radially outer side to the radially inner side. The first wall is connected to the mounting base and provides the fluid channel inlet. The second wall and the first wall form a closed chamber, which provides the fluid channel. The outer wall of the mounting boss provides the third wall. The third wall, the second wall, the base, and the other portion of the membrane body enclose the closed pressure-changing space.

[0010] In one or more embodiments of the culture device, the membrane connection portion includes a protrusion extending axially along the circumferential edge of the membrane body, the top surface of the closed chamber and the second wall surface and the first wall surface form a first connecting ring groove, and the protrusion is matched and connected to the first connecting ring groove; preferably, the membrane connection portion is integrally formed with the membrane body to form the culture membrane.

[0011] In one or more embodiments of the culture apparatus, the surface roughness of the top surface of the mounting boss satisfies Ra less than or equal to 12.5 μm.

[0012] In one or more embodiments of the culture device, the culture unit further includes a fixing part, the base is connected to the mounting seat so that the culture unit is correspondingly mounted on the mounting seat, and the culture membrane is fixed to the base by the fixing part; preferably, the fixing part includes a pressure cap, the pressure cap and the base respectively press and fix the membrane connection part of the culture membrane on both axial sides, a portion of the membrane body corresponding to the top surface of the mounting boss maintains its shape attached to the top surface of the mounting boss during culture, including maintaining the shape of the plane; another portion of the membrane body corresponding to the pressure space deforms within the range of the pressure space during culture; more preferably, the pressure cap is a hollow part, the hollow hole of the pressure cap can be non-sealed and covered by a cover plate so that the culture membrane is completely covered; the pressure cap has a second connecting ring groove, the second connecting ring groove matches the shape of the membrane connection part to press and fix the membrane connection part.

[0013] In one or more embodiments of the culture device, the fluid pressure supply assembly includes a gas source assembly, which includes a fluid pump and a plurality of pressure regulating valves. The fluid pump is fluidly connected to the pressure transformation space, and each pressure regulating valve is respectively disposed in each independent pressure transformation space corresponding to the flow path of the fluid pump. Preferably, the pressure regulating valve is an electro-proportional valve, and a vacuum energy storage element and / or a pressure sensor are also disposed in the flow path.

[0014] In one or more embodiments of the culture device, the fluid pump is connected to the base via a pipeline, the pipeline including a first section adjacent to the base, the first section being a semi-permeable membrane anti-condensation pipe.

[0015] A culture unit according to a second aspect of this application includes: a base, a culture membrane, and a fixing part; the base is connected to a mounting seat so that the culture unit is mounted on the mounting seat, the culture membrane is fixed to the base by the fixing part, and the base has a pressure-changing space connected to the culture membrane, wherein the pressure change of the pressure-changing space causes the culture membrane to undergo flexible deformation; preferably, the culture membrane includes a membrane body and a membrane connecting part located around the membrane body, the base includes a mounting boss, the top surface of the mounting boss is disposed corresponding to a portion of the membrane body, and the pressure-changing space is located on the mounting boss. The outer periphery corresponds to another part of the membrane body; this other part of the membrane body is located on the periphery of one part of the membrane body; more preferably, the base has a first wall, a second wall, and a third wall distributed sequentially from the radially outer side to the radially inner side, the first wall is connected to the mounting base and provides a fluid channel inlet; the second wall and the first wall form a closed chamber, which provides the fluid channel; the outer wall of the mounting boss provides the third wall, and the third wall, the second wall, the base, and the other part of the membrane body enclose the closed pressure transformation space.

[0016] A culture membrane according to a third aspect of this application includes a membrane body and a membrane connection portion located on the periphery of the membrane body, the membrane connection portion including a protrusion extending axially along the circumferential edge of the membrane body; preferably, the culture membrane has no connection holes.

[0017] The beneficial effects of the above embodiments include, but are not limited to, one or a combination of the following:

[0018] By using a mounting base with multiple mutually isolated mounting seats, each mounting seat corresponding to a single culture unit, the multiple culture units of the culture assembly are isolated from each other, and the pressure change spaces of the multiple culture units are independent of each other. This allows the pressure provided by the pressure supply assembly to be precisely adjusted for each culture unit, such as the individual control of the stretching frequency and tensile force. This enables comparative experiments under different conditions to be carried out in a single culture, improving the utilization efficiency of the culture equipment.

[0019] The structure, which uses a fixing part and a base to press and connect the membrane connection part around the periphery of the culture membrane, and an mounting boss to attach and connect the membrane body in the middle part of the culture membrane, achieves a seal for the culture membrane itself. This allows the culture membrane to be stably fixed without the need for screws / bolts, and ensures uniform stress and deformation during the culture process. Furthermore, during the replacement of the culture membrane, the fixing part and the base can be separated to release the fixation of the culture membrane, making the culture membrane easy to replace.

[0020] Overview of the attached figures

[0021] The above and other features, properties and advantages of this application will become more apparent from the following description taken in conjunction with the accompanying drawings and embodiments, wherein:

[0022] Figure 1 is an exploded structural diagram of a cell biomimetic culture device according to an embodiment.

[0023] Figure 2 is a schematic diagram of the structure of the culture component of a cell biomimetic culture device according to an embodiment.

[0024] Figure 3 is an exploded structural diagram of the culture unit of a cell biomimetic culture device according to an embodiment.

[0025] Figures 4A and 4B are schematic diagrams of the deformed and non-deformed structures of the culture membrane of the culture unit of a cell biomimetic culture device according to an embodiment.

[0026] Figure 5 is an exploded structural diagram of the culture unit of a cell biomimetic culture device according to another embodiment.

[0027] Figure 6 is a schematic diagram of the deformation of the culture membrane in the culture unit of another embodiment of the cell biomimetic culture device.

[0028] Figure 7 is a schematic diagram of the gas source component of a cell biomimetic culture device according to an embodiment.

[0029] Figure 8 is a schematic block diagram of the gas source component of a cell biomimetic culture device according to an embodiment.

[0030] Reference numerals: 100-Bionic cell culture equipment; 10-Cultivation component; 20-Fluid pressure supply component; 200-Gas source component; 300-Control component; 1-Cultivation unit; 11-Base; 111-Pressure-changing space; 112-Fluid channel; 1121-Fluid channel inlet; 113-Mounting boss; 1131-Top surface of mounting boss; 1132-Bottom of mounting boss; 114-First connecting part; 115-First wall surface; 116-Second wall surface; 117-Third wall surface; 118-First connecting ring groove; 119-Quick connector; 12-Cultivation membrane; 121-Membrane body; 122-Membrane connecting part; 1220-Protrusion; 123-Connecting hole; 13-Fixing part; 131-Pressure cap; 1310-Hollow hole; 1311-Cover plate; 1312-Second connecting ring groove; 1313-Platform; 2-Mounting base, 21-Mounting seat, 22-Pressure plate, 23-Base, 24-Hinge, 25-Pressure nut, 26-Handle, 27-First section of pipeline; 201-Fluid pump, 202-Pressure regulating valve, 203-Vacuum energy storage element, 204-Pressure sensor.

[0031] Preferred embodiments of the present invention

[0032] The following discloses various implementations or embodiments of the described subject matter. To simplify the disclosure, specific examples of elements and arrangements are described below. These are merely examples and are not intended to limit the scope of protection of the invention. For instance, a first feature formed above or on a second feature, as described later in the specification, can include implementations where the first and second features are formed in a direct connection, or implementations where an additional feature is formed between the first and second features, thus the first and second features may not be directly connected. Furthermore, reference numerals and / or letters may be repeated in different examples in these disclosures. This repetition is for brevity and clarity and does not in itself indicate a relationship between the various implementations and / or structures to be discussed. Further, when the first element is described in connection with or combined with a second element, the description includes implementations where the first and second elements are directly connected or combined with each other, as well as implementations where one or more other intervening elements are added to indirectly connect or combine the first and second elements with each other.

[0033] Referring to Figures 1 to 8, the cell biomimetic culture device 100 includes a culture component 10 and a fluid pressure supply component 20. The culture component 10 is fluidly connected to the fluid pressure supply component 20. The fluid pressure supply component 20 provides the culture component 10 with varying forces, thereby applying external forces to stretch or compress the cells during the cell culture process, simulating the real growth environment of cells in vivo, and performing biomimetic culture.

[0034] As shown in Figure 1, the fluid pressure supply component 20 may include a gas source component 200 and a control component 300. The control component 300 and the gas source component 200 are electrically connected. The control component 300 sends and / or receives analog signals to control the gas source component 200. The gas source component 200 is connected to the culture component 10 through a pipeline, and the gas source component 200 provides the culture component 10 with gases of different pressures and / or frequencies. It can be understood that the three components can be distributed or integrated into a single housing, without limitation. For example, the culture component 10 can be located in an incubator, while the gas source component 200 and the control component 300 can be located outside the incubator. The control component 300 can also remotely and wirelessly connect to control the gas source component 200. Alternatively, the three components can be integrated into the same space defined by the housing, without limitation.

[0035] Referring to Figures 2 to 6, the culture assembly 10 includes multiple culture units 1, which are isolated from each other.

[0036] Referring to Figures 3, 4A, 4B, 5, and 6, the specific structure of culture unit 1 is described in detail below. Culture unit 1 includes a base 11 and a culture membrane 12. The base 11 has a pressure-changing space 111 connected to the culture membrane 12, and the pressure change in the pressure-changing space 111 causes the culture membrane 12 to undergo flexible deformation. The pressure-changing spaces 111 of the multiple culture units 1 are independent of each other. Correspondingly, the fluid pressure supply assembly 20 is fluidly connected to each of the independent pressure-changing spaces 111 to provide pressure changes to the pressure-changing space 111.

[0037] The above mutual isolation and independence refer to the fact that the variable pressure spaces 111 of multiple culture units 1 are not fluidly connected.

[0038] The base 11 can be made of a transparent material, such as polymethyl methacrylate or polycarbonate, but is not limited thereto. The culture membrane 12 can be made of transparent silicone rubber or polyurethane, for example, modified or unmodified polydimethylsiloxane. The culture membrane is flexible and elastic, allowing cells attached to the membrane to deform in a controlled and reproducible manner, simulating cell stretching or compression in the human body to achieve biomimetic culture. Before use, the culture membrane can be sterilized and / or pre-treated with coating as needed. Sterilization can be achieved through high-temperature sterilization or ultraviolet sterilization. Coating pre-treatment can be a conventional protein coating procedure known in the art.

[0039] Referring again to Figure 2, in some embodiments, the culture assembly 10 further includes a mounting base 2 having multiple mounting seats 21, each corresponding to a single culture unit 1. The structure in which the base 11 and the mounting seats 21 are fixedly connected can be, for example, a screw and nut connection, a snap-fit ​​connection, etc.

[0040] In some embodiments, the specific structure of the mounting base 2 can be referred to as shown in Figure 2. The mounting base 21 takes the multiple openings of the mounting base 2 as an example, but it is not limited to this. The number and arrangement of the openings can be set according to the needs of cultivation, such as four, five, or six. Preferably, as shown in Figure 2, the six openings are set at equal intervals.

[0041] In some embodiments, the structure for fixing the mounting base 2 to the culture unit 1 may include a mounting base 2 comprising a clamping plate 22, a base 23, a clamping mechanism, and a hinge 24. The clamping plate 22 and the base 23 may be of the same arbitrary shape, for example, as shown in Figure 2, both the clamping plate 22 and the base 23 are rectangular. The clamping plate 22 and the base 23 are provided with multiple openings at corresponding positions as mounting seats 21.

[0042] The clamping plate 22 and the base 23 can be connected by a hinge 24. The openings in the clamping plate 22 and the base 23 are used to house the culture units 1, with each culture unit 1 corresponding to one of the openings. The clamping plate 22 and the base 23 interact to clamp the culture units, thus fixing them in place. In some embodiments, the clamping plate 22, the culture units 1, and the base 23 can be fixed by a clamping mechanism. The clamping mechanism can include clamping nuts 25, snap-fit ​​clamps, spring clamps, and other common clamping mechanisms in the art, which will not be described in detail here. In some embodiments, the clamping mechanism is a clamping nut 25. Additionally, a handle 26 can be provided on the clamping plate 22 to further facilitate the disassembly and replacement of the culture units. In some embodiments, the clamping plate 22 and the base 23 can also be separated, i.e., not connected by a hinge 24, but fixed solely by a clamping mechanism.

[0043] Referring again to Figures 3 to 6, in some embodiments, the base 11 has a fluid channel 112, and the pressure-changing space 111 is connected to the fluid channel 112. The fluid channel inlets 1121 of the respective fluid channels 112 of the multiple culture units 1 are independent of each other. The structure of integrating the fluid channel 112 on the base 11 achieves independence between the pressure-changing spaces 111 of the multiple culture units 1 through a simple structure, and the integration of the structure is good. The fluid channel inlet 1121 can be connected to a quick connector 119. A quick connector 119 is provided on the radial outer side wall of the base 11. The quick connector 119 is connected to the fluid channel inlet 1121 and thus communicates with the fluid channel 112. The quick connector 119 is used to connect a gas pipeline to provide pressure changes within the pressure-changing space 111. In some embodiments, the gas pipeline includes a first section 27 adjacent to the base 11. The first section 27 is a semi-permeable membrane anti-condensation pipe. The remaining sections of the gas pipeline can be ordinary gas pipelines, such as polyurethane hoses, stainless steel pipes, etc. The principle of the semi-permeable membrane anti-condensation pipe is that when there is a difference in humidity between the inside and outside of the pipe, water vapor will permeate from the side with higher humidity to the side with lower humidity. The anti-condensation pipe uses a semi-permeable membrane, so only water vapor can pass through, and almost no air can pass through. The mist generated each time the gas is exhausted causes the humidity inside the pipe to increase, which may lead to condensation. The anti-condensation pipe allows the mist generated inside the pipe to permeate from the pipe with higher humidity to the outside of the pipe with lower humidity, thereby preventing water vapor from accumulating inside the pipe and thus preventing condensation. The beneficial effect of using the anti-condensation pipe is that the inventors discovered that the gas delivered by the gas source component and the temperature of the culture component during the culture process are different, so condensate will be generated during the culture process. Using the anti-condensation pipe, the condensate can be discharged incidentally through the pressure change process of biomimetic culture, and water vapor can also be discharged through the anti-condensation pipe, thereby further optimizing the humidity conditions in the culture environment.

[0044] Referring again to Figures 3 to 6, in some embodiments, the culture membrane 12 may include a membrane body 121 and a membrane connector 122 located around the membrane body 121. In some embodiments, the base 11 includes a mounting boss 113, the top surface 1131 of which corresponds to a portion of the membrane body 121, and a pressure-changing space 111 located around the mounting boss 113, corresponding to another portion of the membrane body 121; this other portion of the membrane body 121 is located around the portion of the membrane body 121. It can be understood that, generally, most of the membrane body 121 corresponds to the top surface 1131 of the mounting boss, while a small portion of the membrane body 121 corresponds to the pressure-changing space 111. For example, 50% to 95% of the area of ​​the membrane body 121 may correspond to the mounting boss 113, while 5% to 50% of the area of ​​the membrane body may correspond to the pressure-changing space 111. The shape of the mounting boss can be any shape, such as the circle shown in the figure, but is not limited thereto. For example, it can also be rectangular, square, elliptical, or irregular, as well as a combination of multiple shapes, without limitation. The beneficial effect is that, through the cooperative relationship of the mounting boss 113, the membrane body 121, and the membrane connecting part 122, a reliable, stable, and uniformly stressed fixed connection of the membrane body 121 can be achieved with a simple structure without threaded connection holes or a threaded connection structure. The dashed connection hole 123 shown in Figure 3 is only to exemplify the approximate location of the threaded connection hole 123 in the comparative scheme. The culture membrane in the comparative scheme is generally a planar structure with only the connection hole 123, without the protrusion 1220. In this embodiment, the culture membrane 12 does not have the connection hole 123 and is not connected and fixed to the base 11 through a threaded connection structure. Instead, the membrane connecting part 122, exemplified by the protrusion 1220, is directly pressed and fixed by the base 11 and the fixing member 13. This further optimizes the structural stability and uniformity of stress of the culture membrane during the culture process. In some embodiments, the surface roughness of the top surface 1131 of the mounting boss satisfies Ra less than or equal to 12.5 μm. The inventors have found that when the top surface 1131 of the mounting boss meets this roughness requirement, the stress uniformity of the culture membrane 12 is significantly optimized.

[0045] Referring again to Figures 3 to 4B, in some embodiments, the base 11 may further include a first connecting portion 114 corresponding to the membrane connecting portion 122. The radial gap between the first connecting portion 114 and the mounting boss 113 provides a pressure-changing space 111, thereby improving the integration of the base 11. Specifically, in some embodiments, the base 11 may have a first wall surface 115, a second wall surface 116, and a third wall surface 117 distributed sequentially from the radially outer side to the radially inner side. The first wall surface 115 is connected to the mounting base 21 and provides a fluid channel inlet 1121. The second wall surface 116 and the first wall surface 115 form a closed chamber. Here, "closed" means that it is only connected to the outside through the fluid channel inlet and outlet, while the rest is closed. The enclosed chamber provides a fluid channel 112; the outer wall of the mounting boss 113 provides a third wall 117. The third wall 117, the second wall 116, the base 11, and another part of the membrane body 121 enclose a closed pressure-changing space 111. Similarly, the "enclosed" here refers to the fact that it is only connected to the fluid channel outlet, while the rest is closed. The second wall 116 and the first wall 115 form a closed chamber, and the third wall 117, the second wall 116, the base 11, and another part of the membrane body 121 enclose a closed pressure-changing space 111, as shown in Figures 3 to 4B. It can be a complete ring, or one or more parts of a ring, semicircle, or rectangle, without limitation. The second wall 116 and the third wall 117 can be completely separated or partially separated, that is, the second wall 116 and the third wall 117 share a portion. The use of a complete ring structure is beneficial to optimizing the stress uniformity of the culture membrane and the stability of the fixation structure. It can be understood that the structure providing the transformer space 111 can also be as shown in Figures 5 and 6. In some embodiments, unlike the bottom of the mounting boss 113 shown in Figures 3 to 4B which is completely connected to the base 11, in the embodiments shown in Figures 5 and 6, the bottom of the mounting boss 113 is partially connected to the base 11, so that the groove formed between the bottom 1132 of the mounting boss and the base 11 provides part of the transformer space 111, thus further increasing the transformer space 111.

[0046] Referring again to Figures 3 to 4B, in some embodiments, the specific structure for connecting the membrane connector 122 to the base 11 may include a protrusion 1220 extending axially from at least one side of the circumferential edge of the membrane body 121. The thickness of the membrane body 121 may be 0.4 mm to 0.6 mm, and the protrusion length of the protrusion 1220 may be 0.8 mm to 15 mm, preferably 0.8 mm to 5 mm. The top surface of the closed chamber, the second wall surface 116, and the first wall surface 115 may form a first connecting annular groove 118, with the protrusion 1220 matching and connecting to the first connecting annular groove 118, thus further optimizing the stability of the connection structure. In some embodiments, the membrane connector 122 and the membrane body 121 are integrally connected to form a culture membrane 12, for example, by injection molding to form an integrally connected structure, but this is not a limitation. An integrally connected structure provides better uniform stress distribution.

[0047] Referring again to Figures 3, 4A, and 4B, in some embodiments, the culture unit 1 may further include a fixing part 13. The base 11 is connected to the mounting base 21, so that the culture unit is mounted on the mounting base 21, and the culture membrane is fixed to the base 11 by the fixing part 13. In some embodiments, the structure of the fixing part 13 may include a pressure cap 131. The pressure cap 131 and the base 11 respectively press and fix the membrane connection part 122 of the culture membrane 12 on both axial sides. A portion of the membrane body 121 corresponding to the top surface 1131 of the mounting boss remains attached to the shape of the top surface 1131 of the mounting boss during culture. Specifically, it may be a shape including a plane as shown in Figures 4A and 4B. Another portion of the membrane body 121 provided in the pressure space 111 deforms within the range of the pressure space 111 during culture.

[0048] The shapes of the base 11, culture membrane 12 and fixing seat 13 described above are not limited to the circles shown in the figure. The three can be any shape with the same shape, such as rectangles, squares, pentagons and other shapes, as well as combinations of multiple shapes. In one embodiment, the base 11, culture membrane 12 and fixing seat 13 are circular in shape, which can make the force more uniform.

[0049] Referring to Figures 4A and 4B, the cap 131 is a hollow component. The hollow hole 1310 of the cap can be non-sealed by the cover plate 1311. Non-sealed coverage means that although the culture membrane 12 is completely covered in the figures, it is not tightly sealed. Air can enter and exit through gaps during the culture process. Therefore, during culture, a sealed structure is formed between the lower surface of the culture membrane body and the pressure-changing space, while the space on the upper surface of the membrane body for cell inoculation is connected to the external gas, simultaneously preventing contamination of the culture membrane 12. Continuing to refer to Figures 4A and 4B, in some embodiments, the cap 131 has a second connecting ring groove 1312. The second connecting ring groove 1312 matches the shape of the membrane connection portion 122 to press and fix the membrane connection portion 122, thus further optimizing the stability and uniformity of the fixation of the culture membrane 12.

[0050] Referring again to Figures 3 to 6, the method for installing and fixing the culture membrane 12 may include the following steps: Lubricating oil is evenly applied to the mounting protrusion 113 of the base 11 to ensure smooth lubrication between the mounting protrusion 113 and the culture membrane 12. The lubricating oil is a silicone-based lubricating oil, preferably a transparent non-curing silicone grease. The lower part of the protrusion 1220 of the culture membrane 12 is inserted into the first connecting part 114 of the corresponding membrane connecting part 122 of the groove structure. A pressure cap 131 is placed above the protrusion 1220 of the culture membrane. The pressure cap 131 has a second connecting ring groove 1312, the shape of which matches the protrusion 1220 to press and fix the membrane connecting part 122. The pressure cap also includes a platform 1313 extending outward from the second connecting ring groove 1312. A clamping plate 22 presses down on the platform 1313 and tightens the clamping nut to press the culture membrane 12 onto the base 11. The culture membrane is flexible and elastic. After compression, the culture membrane 12 and the base 11 form a closed cavity to provide a pressure-changing space 111.

[0051] During cell culture, the biomimetic cell culture device can provide tensile or compressive forces. When tensile force is required, as shown in Figure 4B, cells are seeded on the culture membrane 12 at positions corresponding to the protrusions 113 under normal pressure in the variable pressure space 111. The gas source component 200 applies negative pressure, causing the culture membrane 12 to deform, as shown in Figure 4A or Figure 6. The membrane curves concave within the variable pressure space 111, while the middle portion remains horizontal due to the support of the protrusions 113. However, the corresponding middle portion of the culture membrane has undergone tensile deformation, thus achieving cell culture under tensile force.

[0052] Similarly, when compressive force culture is required, cells are seeded on the culture membrane at positions corresponding to the protrusions 113 under negative pressure conditions in the variable pressure space 111. When the gas source system returns to normal pressure, the culture membrane 12 contracts, and the adherent cells on the culture membrane 12 are subjected to compressive force as the culture membrane contracts, thus achieving cell compressive force culture.

[0053] In some embodiments, both the base 11 and the culture membrane 12 can be made of transparent material, so that the cell biomimetic culture device can be adapted to both upright and inverted microscopes to observe the stretching culture process in real time.

[0054] Referring to Figure 7, in some embodiments, the fluid pressure supply assembly 20 includes a gas source assembly 200, which includes a fluid pump 201 and multiple pressure regulating valves 202. The fluid pump 201 is fluidly connected to the pressure changing space 111, and each pressure regulating valve 202 is respectively disposed in the flow path of each independent pressure changing space 111 corresponding to the fluid pump 201. Each culture unit 1 corresponds to one pressure regulating valve 202, and the pressure of each culture unit can be controlled independently. Preferably, the pressure regulating valve 202 is an electro-proportional valve. The inventors have found that, compared with the commonly used combination of directional valves and proportional valves, the control logic of the electro-proportional valve is simpler. Furthermore, the inventors have found that the electro-proportional valve also meets the service life requirements under the conditions of frequent pressure changes in biomimetic culture, and the use of the electro-proportional valve can further meet the requirements of precise pressure control and uniform force on the culture membrane 12.

[0055] The fluid pump 201 may be a vacuum pump, for example, to provide the required pressure to the culture unit 1. In some embodiments, a vacuum energy storage element 203, such as a vacuum energy storage tank, may also be provided between the fluid pump 201 and the pressure regulating valve 202. The vacuum energy storage tank is capable of storing vacuum and balancing system pressure.

[0056] As shown in Figure 8, the control component 300 can change the output pressure of the vacuum pressure regulating valve 202 according to different analog inputs. Each vacuum pressure regulating valve 202 can have a pressure sensor 204 installed downstream to provide feedback to the control component 300, comparing the actual pressure with the input pressure. The control component 300 can be a multi-channel controller, with each channel corresponding to a separate vacuum pressure regulating valve 202. The control component 300 can achieve different waveforms, frequencies, and pressure values ​​based on the different analog input waveforms, frequencies, and values ​​of each channel through the vacuum pressure regulating valves. By applying different pressure values ​​and curves to different cavities, different human body environments can be simulated, corresponding to various cellular morphologies. For example, the waveform can be a static waveform, rectangular waveform, triangular waveform, sine waveform, cardiac waveform, etc.; the pressure range is -80 kPa to 0 kPa; and the strain frequency is 0 to 5 Hz.

[0057] As described above, this application also provides a culture unit 1, including: a base 11, a culture membrane 12, and a fixing part 13; the base 11 is connected to a mounting base 21 so that the culture unit 1 is mounted on the mounting base 21, the culture membrane 12 is fixed to the base 11 by the fixing part 13, the base 11 has a pressure-changing space 111 connected to the culture membrane 12, and the pressure change of the pressure-changing space 111 causes the culture membrane 12 to undergo flexible deformation; the culture membrane 12 includes a membrane body 121 and a membrane connecting part 122 located around the membrane body 121, the base 11 includes a mounting boss 113, the top surface 1131 of the mounting boss is disposed corresponding to a portion of the membrane body 121, and the pressure-changing space 111 is located at the mounting base 111. The outer periphery of the boss 113 corresponds to another part of the membrane body 121; this other part of the membrane body 121 is located on the periphery of one part of the membrane body 121; the base 11 has a first wall surface 115, a second wall surface 116, and a third wall surface 117 distributed sequentially from the radially outer side to the radially inner side. The first wall surface 115 is connected to the mounting base 21 and provides a fluid channel inlet 1121; the second wall surface 116 and the first wall surface 115 form a closed chamber, which provides a fluid channel 112; the outer wall surface of the mounting boss 113 provides a third wall surface 117, and the third wall surface 117, the second wall surface 116, the base 11, and the other part of the membrane body 121 enclose a closed pressure transformation space 111.

[0058] This application also provides a culture membrane 12, including a membrane body 121 and a membrane connection portion 122 located on the periphery of the membrane body 121. The membrane connection portion 122 includes a protrusion 1220 extending axially along the circumferential edge of the membrane body 121. Preferably, the culture membrane 12 has no connection hole 123.

[0059] In summary, the beneficial effects of the cell biomimetic culture equipment, culture unit, and culture membrane described in the above embodiments include, but are not limited to, one or a combination of the following:

[0060] By using a mounting base with multiple isolated mounting seats, each corresponding to a single culture unit, the multiple culture units of the culture assembly are isolated from each other, and the pressure change spaces of the multiple culture units are independent of each other. This allows the pressure provided by the pressure supply assembly to be precisely adjusted for each culture unit, such as the individual control of the stretching frequency and tensile force. This enables comparative experiments under different conditions to be carried out in a single culture, improving the efficiency of the culture equipment.

[0061] The structure, which uses a fixing part and a base to press and connect the membrane connection part around the periphery of the culture membrane, and an mounting boss to attach and connect the membrane body in the middle part of the culture membrane, achieves a seal for the culture membrane itself. This allows the culture membrane to be stably fixed without the need for screws / bolts, and ensures uniform stress and deformation during the culture process. Furthermore, during the replacement of the culture membrane, the fixing part and the base can be separated to release the fixation of the culture membrane, making the culture membrane easy to replace.

[0062] While this application discloses preferred embodiments as described above, it is not intended to limit the scope of this application. Any changes and modifications can be made by those skilled in the art without departing from the spirit and scope of this application. Therefore, any modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of this application without departing from the content of the technical solution of this application shall fall within the protection scope defined by the claims of this application.

Claims

1. A cell biomimetic culture device (100), characterized in that, include: The culture component (10) includes multiple culture units, which are isolated from each other; The culture unit (1) includes a base (11) and a culture membrane (12); the base (11) has a pressure-changing space (111) connected to the culture membrane (12), and the culture membrane (12) undergoes flexible deformation due to pressure changes in the pressure-changing space (111); wherein the pressure-changing spaces (111) of the plurality of culture units (1) are independent of each other; A fluid pressure supply assembly (20) is fluidly connected to each of the independent pressure-changing spaces (111) to provide pressure changes in the pressure-changing spaces (111).

2. The cell biomimetic culture device (100) as described in claim 1, characterized in that, The culture assembly (10) further includes a mounting base (2), which has a plurality of mounting seats (21), each of which corresponds to a single culture unit; Preferably, the base (11) has a fluid channel (112), and the pressure-changing space (111) is connected to the fluid channel (112); the fluid channels (1121) of the respective fluid channels (112) of the multiple culture units (1) are independent of each other.

3. The cell biomimetic culture device (100) as described in claim 2, characterized in that, The culture membrane (12) includes a membrane body (121) and a membrane connector (122) located around the membrane body (121). The base (11) includes a mounting boss (113). The top surface (1131) of the mounting boss (113) is disposed corresponding to a portion of the membrane body (121). The pressure-changing space (111) is located around the mounting boss (113) and is disposed corresponding to another portion of the membrane body (121). The other portion of the membrane body (121) is located around the portion of the membrane body (121). Preferably, the base (11) includes a first connecting portion (114) corresponding to the membrane connecting portion (122), and the radial gap between the first connecting portion (114) and the mounting boss (113) provides the pressure transformation space (111); More preferably, the base (11) has a first wall (115), a second wall (116), and a third wall (117) distributed sequentially from the radially outer side to the radially inner side. The first wall (115) is connected to the mounting base (21) and provides the fluid channel inlet (1121). The second wall (116) and the first wall (115) form a closed chamber, which provides the fluid channel (112). The outer wall of the mounting boss (113) provides the third wall (117). The third wall (117), the second wall (116), the base (11), and the other part of the membrane body (121) enclose the closed pressure transformation space (111).

4. The cell biomimetic culture device (100) as described in claim 3, characterized in that, The membrane connection part (122) includes a protrusion (1220) extending axially along the circumferential edge of the membrane body (121). The top surface of the closed chamber, the second wall surface (116), and the first wall surface (115) form a first connecting annular groove (118). The protrusion (1220) is matched and connected to the first connecting annular groove (118). Preferably, the membrane connector (122) is integrally formed with the membrane body (121) to form the culture membrane (12).

5. The cell biomimetic culture device (100) as described in claim 3, characterized in that, The surface roughness of the top surface (1131) of the mounting boss satisfies Ra less than or equal to 12.5 μm.

6. The cell biomimetic culture device (100) as described in claim 3, characterized in that, The culture unit (1) further includes a fixing part (13), the base (11) is connected to the mounting base (21) so that the culture unit is installed on the mounting base (21) and the culture membrane is fixed to the base (11) by the fixing part (13); Preferably, the fixing part (13) includes a pressure cap (131), the pressure cap (131) and the base (11) respectively press and fix the membrane connection part (122) of the culture membrane (12) on both sides of the axial direction, and a part of the membrane body (121) corresponding to the top surface (1131) of the mounting boss maintains its shape attached to the top surface (1131) of the mounting boss during culture, including maintaining the shape of the plane; another part of the membrane body (121) corresponding to the pressure space (111) deforms within the range of the pressure space (111) during culture. More preferably, the cap (131) is a hollow part, and the hollow hole (1310) of the cap can be non-sealed by the cover plate (1311) so that the culture membrane (12) is completely covered; the cap (131) has a second connecting ring groove (1312), and the second connecting ring groove (1312) matches the shape of the membrane connection part (122) to press and fix the membrane connection part (122).

7. The cell biomimetic culture device (100) as described in claim 1, characterized in that, The fluid pressure supply assembly (20) includes a gas source assembly (200), which includes a fluid pump (201) and a plurality of pressure regulating valves (202). The fluid pump (201) is fluidly connected to the pressure transformation space (111), and each pressure regulating valve (202) is respectively provided in each independent pressure transformation space (111) and is connected to the flow path of the fluid pump (201). Preferably, the pressure regulating valve (202) is an electro-proportional valve, and a vacuum energy storage element (203) and / or a pressure sensor (204) are also provided in the flow path.

8. The cell biomimetic culture device (100) as described in claim 7, characterized in that, The fluid pump (201) is connected to the base (11) through a pipeline, the pipeline including a first section (27) adjacent to the base (11), the first section (27) being a semi-permeable membrane anti-condensation pipe.

9. A culture unit (1), characterized in that, include: The base (11), culture membrane (12), and fixing part (13) are provided. The base (11) is connected to the mounting base (21) so that the culture unit (1) is installed on the mounting base (21). The culture membrane (12) is fixed to the base (11) by the fixing part (13). The base (11) has a pressure-changing space (111) connected to the culture membrane (12). The pressure change of the pressure-changing space (111) causes the culture membrane (12) to undergo flexible deformation. Preferably, the culture membrane (12) includes a membrane body (121) and a membrane connector (122) located around the membrane body (121). The base (11) includes a mounting boss (113). The top surface (1131) of the mounting boss (113) is disposed corresponding to a portion of the membrane body (121). The pressure-changing space (111) is located around the mounting boss (113) and is disposed corresponding to another portion of the membrane body (121). The other portion of the membrane body (121) is located around the portion of the membrane body (121). More preferably, the base (11) has a first wall (115), a second wall (116), and a third wall (117) distributed sequentially from the radially outer side to the radially inner side. The first wall (115) is connected to the mounting base (21) and provides a fluid channel inlet (1121). The second wall (116) and the first wall (115) form a closed chamber, which provides the fluid channel (112). The outer wall of the mounting boss (113) provides the third wall (117). The third wall (117), the second wall (116), the base (11), and the other part of the membrane body (121) enclose the closed pressure transformation space (111).

10. A culture membrane (12), characterized in that, The membrane includes a membrane body (121) and a membrane connection portion (122) located around the membrane body (121). The membrane connection portion (122) includes a protrusion (1220) extending axially along the circumferential edge of the membrane body (121). Preferably, the culture membrane (12) has no connection hole (123).

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