Microfluidic chip for building modular cell cultures and method of fabrication

Abstract

This invention discloses a microfluidic chip and its fabrication method for constructing modular cell cultures, relating to the field of cell culture technology. The chip includes an upper plate and a lower plate. The upper plate includes a first flow channel and a second flow channel, while the lower plate includes multiple spaced-apart culture chambers, each with a different cross-sectional shape. This allows each chamber to be independently formed into cultures of different shapes. The cultures can be stacked and combined after removal to construct a multi-cell co-culture model with functional partitions, significantly improving structural complexity and biomimicry. The upper plate slides to switch the connection between the first and second flow channels and the culture chambers, enabling inoculation and culture modes. During inoculation, cultured cells are injected into the culture chamber through the first flow channel; during culture, the flow channel is switched to perfuse the culture chamber with culture medium, avoiding damage to the culture during residual gel removal. This simple operation improves the success rate and enhances the reliability of experimental results.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and in particular to a microfluidic chip for constructing modular cell cultures and a processing method thereof. Background Technology

[0002] Organoid technology, as a novel in vitro modeling technology, has shown significant advantages in mechanism research and drug testing. However, traditional organoid models have relatively limited cell types, making it difficult to recreate the complex multi-cell interactions in vivo. Current co-culture methods, which directly mix organoids and cells in hydrogels or culture media, are simple to operate but struggle to reproduce the functional partitioning between different cells in vivo, and cannot construct large-scale functional structures. While 3D bioprinting technology can achieve precise cell positioning in three-dimensional space, it is limited by the choice of materials and cells for printing equipment, and is particularly difficult to adapt to organoid culture systems that primarily use temperature-sensitive materials such as matrix gels and decellularized extracellular matrix. The additional excipients introduced increase experimental variables and uncertainty in the results. Summary of the Invention

[0003] This invention aims to at least solve one of the technical problems existing in the prior art. To this end, this invention proposes a microfluidic chip and processing method for constructing modular cell cultures. By constructing cell cultures of different shapes, and by assembling a multi-cell culture model using multiple different shaped cultures, the construction of multi-cell functional differentiation regions can be achieved.

[0004] A microfluidic chip for constructing modular cell cultures according to a first aspect embodiment of the present invention includes: The upper plate is equipped with a first flow channel and a second flow channel; The lower plate is provided with multiple spaced-apart culture chambers, and each of the multiple culture chambers has a different cross-sectional shape along the horizontal direction, so as to shape cultures with different shapes. The upper plate is slidably connected to the lower plate. When the first flow channel and the culture chamber overlap, they can communicate with each other, so that inoculation can be performed into the culture chamber through the first flow channel. When the second flow channel overlaps with the culture chamber, the second flow channel can be circulated with culture medium to cultivate the culture in the culture chamber; The cultures with different shapes can be stacked together to form a culture model.

[0005] A microfluidic chip for constructing modular cell cultures according to an embodiment of the present invention has at least the following beneficial effects: This embodiment includes a slidingly connected upper plate and a lower plate. The upper plate includes a first flow channel and a second flow channel, and the lower plate includes multiple spaced-apart culture chambers, each with a different cross-sectional shape. This allows each culture chamber to be independently formed into cultures of different shapes, and the cultures can be stacked and combined after removal to construct a multi-cell co-culture model with functional partitions, significantly improving the complexity and biomimicry of the structure. The upper plate slides to switch the connection between the first and second flow channels and the culture chambers, thereby achieving inoculation and culture modes respectively. During inoculation, the first flow channel injects cultured cells into the culture chamber; during culture, the second flow channel perfuses the culture chamber with culture medium, avoiding damage to the culture during residual gel removal. This simplifies operation, improves the success rate, and enhances the stability of the culture system and the reliability of experimental results.

[0006] According to an embodiment of the first aspect of the present invention, the lower plate is provided with a plurality of spaced-apart culture zones, each culture zone having a plurality of culture chambers, and each culture chamber in each culture zone having a different cross-sectional shape along the horizontal direction.

[0007] According to an embodiment of the first aspect of the present invention, the first flow channel is provided with a plurality of spaced-apart connecting cavities. When the first flow channel overlaps with the plurality of culture cavities, the plurality of connecting cavities are arranged alternately with the plurality of culture cavities, and two adjacent connecting cavities can simultaneously connect to the culture cavities located between them, so that the two adjacent culture cavities can be interconnected, thereby enabling inoculation into the culture cavities through the first flow channel.

[0008] According to an embodiment of the first aspect of the present invention, the culture region includes a first region and a second region, and the culture chambers of both regions include a base and a protrusion that communicate with each other. The protrusion extends outward from one side of the base. The protrusion of the culture chamber in the first region communicates with an end of the base, and the protrusion of the culture chamber in the second region communicates with a middle portion of the base.

[0009] According to an embodiment of the first aspect of the present invention, the communicating cavity includes a first communicating portion and a second communicating portion. When the communicating cavity is communicating with the culture cavity, a portion of the first communicating portion can overlap with a portion of the base, and a portion of the second communicating portion can overlap with a portion of the protrusion; and / or, both ends of the first flow channel and the second flow channel are respectively provided with a first injection channel and a second injection channel, the first injection channel being used for injecting cell fluid, and the second injection channel being used for injecting culture medium.

[0010] According to an embodiment of the first aspect of the present invention, the culture zone includes a third region and a fourth region, and the culture chambers of the third region and the fourth region each have a rectangular cross-section, wherein the aspect ratio of the culture chamber in the third region is not equal to that of the culture chamber in the fourth region.

[0011] According to an embodiment of the first aspect of the present invention, the upper plate is provided with a first positioning groove and a second positioning groove on both sides, and the lower plate is provided with a fixing groove. A positioning pin can be inserted between the first positioning groove and the fixing groove so that the first flow channel can communicate with the culture chamber. When the positioning pin is inserted between the second positioning groove and the fixing groove, the second flow channel can communicate with the culture chamber.

[0012] According to an embodiment of the first aspect of the present invention, the system further includes a base, the lower plate being fixed to the base, the base being provided with a pivotally connected positioning member, and the positioning member being provided with the positioning pin.

[0013] According to an embodiment of the first aspect of the present invention, the base is provided with a guide plate that extends upward from the base and is disposed on both sides of the upper plate for guiding the movement of the upper plate.

[0014] According to an embodiment of the second aspect of the present invention, a processing method is provided, applied to a microfluidic chip for constructing modular cell cultures as described in any of the above embodiments, comprising the following steps: The upper plate, lower plate, and base are manufactured and then sterilized. Cover the lower plate with the upper plate, and install the lower plate onto the base; The upper plate is slid to connect the first flow channel to the culture chamber, and is inserted into the first positioning groove and the fixing groove through the positioning pin of the positioning member, so that the upper plate and the lower plate are fixed to each other; The solution containing the cells and precursor material is injected into the first flow channel through the first injection channel to inject into the culture chamber; Remove the positioning component to move the upper plate relative to the lower plate so that the second flow channel connects to the culture chamber, and re-fix the upper plate and the lower plate by inserting the positioning pin into the second positioning groove and the fixing groove; Connect one end of the second injection channel to the perfusion system, and inject culture medium into the second channel through the perfusion system so that the culture medium flows through the culture chamber. At the same time, connect the other end of the second injection channel to the waste liquid tank. After a preset incubation time, the upper plate and the lower plate are separated, the culture formed in the incubation chamber is removed, and the culture is placed in a new incubation environment. Multiple cultures with different shapes are stacked together to form a culture model.

[0015] The processing method according to the second aspect of the present invention has at least the following beneficial effects: In this embodiment, the inoculation mode and culture mode can be switched by sliding the upper plate. The cooperation of the positioning pin with the first positioning groove and the second positioning groove ensures precise alignment and fixation of the flow channel in both modes, avoiding operational deviation. During inoculation, cells and precursor materials can be directly injected into the culture chamber for shaping. After switching to culture mode, the culture medium flows continuously through the culture chamber for dynamic cultivation, eliminating the need to remove residual gel in the flow channel, significantly reducing the risk of culture damage or contamination due to cumbersome operation. After culture is completed, the upper and lower plates can be separated to easily remove the formed modular culture. Finally, multiple cultures with different cross-sectional shapes can be stacked and bonded as needed to flexibly construct a three-dimensional multi-cell co-culture model with functional partitions. This method does not rely on complex printing equipment, is compatible with mainstream temperature-sensitive materials such as matrix adhesives, has a low operation threshold, and is highly versatile.

[0016] 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

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 This is an exploded view of a microfluidic chip for constructing modular cell cultures according to an embodiment of the present invention; Figure 2 This is a first cross-sectional view of the upper and lower plates in the inoculation mode in an embodiment of the present invention; Figure 3 for Figure 2 A magnified view of A in the middle; Figure 4 This is a second cross-sectional view of the upper and lower plates in the inoculation mode in an embodiment of the present invention; Figure 5 for Figure 4 A magnified view of B in the middle; Figure 6 This is a first cross-sectional view of the upper and lower plates in the cultivation mode in an embodiment of the present invention; Figure 7 for Figure 6 A magnified view of C; Figure 8 This is a second cross-sectional view of the upper and lower plates in the cultivation mode in an embodiment of the present invention; Figure 9 This is a schematic diagram of the cultivation model in an embodiment of the present invention; Figure 10 This is a bottom view of the base in an embodiment of the present invention; Figure 11This is a flowchart illustrating the processing method in an embodiment of the present invention.

[0018] Figure label: Base 100; Positioning component 101; Positioning pin 102; Guide plate 103; Cultivation model 104; Observation window 105; Upper plate 110; first flow channel 111; second flow channel 112; first injection channel 113; second injection channel 114; first positioning groove 115; second positioning groove 116; connecting cavity 117; first connecting part 118; second connecting part 119; Lower plate 120; first region 121; second region 122; third region 123; fourth region 124; culture chamber 126; base 127; protrusion 128; fixing groove 129. Detailed Implementation

[0019] Embodiments of the present invention are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0020] In the description of this invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limiting this invention.

[0021] In the description of this invention, "several" means one or more, "more than" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0022] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0023] Reference Figures 1 to 10A microfluidic chip for constructing modular cell cultures, according to a first aspect embodiment of the present invention, includes an upper plate 110, a lower plate 120, and a base 100. The upper plate 110 is slidably connected to the lower plate 120, and the inoculation mode and culture mode can be switched by relative sliding. The base 100 is provided with two movable positioning members 101 for quickly positioning and locking the chip when switching modes.

[0024] Understandably, the lower plate 120 is cuboid in shape, with multiple spaced-apart culture chambers 126 inside. Each culture chamber 126 has a different cross-sectional shape along the horizontal direction, including rounded multi-unit cube structures resembling "Tetris". The different shapes of the culture chambers 126 can be used to shape cell or organoid cultures with different macroscopic morphologies, independently forming cultures with different geometric shapes, providing a structural basis for subsequent construction of functionalized cell partitions.

[0025] Specifically, the lower plate 120 is provided with multiple spaced-apart culture zones, including a first zone 121, a second zone 122, a third zone 123, and a fourth zone 124. Each culture zone has multiple culture chambers 126, and the culture chambers 126 in each culture zone have different cross-sectional shapes along the horizontal direction. For example, the culture chambers 126 in the first zone 121 and the second zone 122 each include a base 127 and a protrusion 128 that are interconnected, with the protrusion 128 extending outward from one side of the base 127. Specifically, the protrusion 128 of the culture chamber 126 in the first zone 121 is connected to the end of the base 127, forming an L-shaped structure; the protrusion 128 of the culture chamber 126 in the second zone 122 is connected to the middle of the base 127, forming a T-shaped structure. The culture chambers 126 in both the third region 123 and the fourth region 124 have rectangular cross-sections, but the aspect ratios of the culture chambers 126 in the third region 123 and the fourth region 124 are not equal. For example, the culture chambers 126 in the third region 123 are elongated strips, while the culture chambers 126 in the fourth region 124 are rectangular or square. These culture chambers 126 with different shapes have geometric features that allow them to be stacked on top of each other, thereby pre-defining the spatial distribution of different cell types in three-dimensional space and solving the problem that existing mixed culture methods cannot construct functional partitions.

[0026] Reference Figure 1 and Figure 5 The upper plate 110 is rectangular and has a first flow channel 111 and a second flow channel 112. Both ends of the first flow channel 111 and the second flow channel 112 are respectively provided with a first injection channel 113 and a second injection channel 114, wherein the first injection channel 113 is used to inject hydrogel precursor material containing cells or organoids, and the second injection channel 114 is used to inject culture medium.

[0027] When the upper plate 110 and the lower plate 120 are combined and in cell seeding mode, the first flow channel 111 and the culture chamber 126 partially overlap on the vertical projection plane. At this time, the first flow channel 111 and the culture chamber 126 are interconnected, so that seeding can be performed on the culture chamber 126 through the first flow channel 111.

[0028] To further improve the efficiency and uniformity of simultaneous inoculation of multiple culture chambers 126, the first flow channel 111 is also provided with multiple spaced-apart connecting chambers 117. When the first flow channel 111 overlaps with the multiple culture chambers 126, the multiple connecting chambers 117 are staggered with the multiple culture chambers 126, and two adjacent connecting chambers 117 can simultaneously connect to the culture chamber 126 located between the two connecting chambers 117, thereby enabling two adjacent culture chambers 126 to communicate with each other, that is, enabling multiple culture chambers 126 to be connected simultaneously. In this way, the cell suspension in the first flow channel 111 can simultaneously fill multiple culture chambers 126 through the connecting chambers 117, avoiding the tedious operation of inoculating one by one, while ensuring the uniformity of cell density in each culture chamber 126.

[0029] Furthermore, the connecting cavity 117 includes a first connecting portion 118 and a second connecting portion 119. When the connecting cavity 117 is connected to the culture cavity 126, a portion of the first connecting portion 118 can overlap with a portion of the base 127, and a portion of the second connecting portion 119 can overlap with a portion of the protrusion 128. This matching design ensures that regardless of whether the shape of the culture cavity 126 is "L", "T" shaped, or other complex shapes, the connecting cavity 117 can cover its main structural area, allowing the hydrogel precursor material to completely fill the entire culture cavity 126, avoiding local material shortages or air bubble residue, thereby improving the integrity of culture formation.

[0030] When the chip is in culture mode, the second flow channel 112 completely covers the lower culture chamber 126 on the vertical projection plane. At this time, the second flow channel 112 can be filled with culture medium to continuously perfuse the culture in the culture chamber 126, which can continuously provide fresh nutrients to the culture and remove metabolic waste, making it closer to the dynamic microenvironment in vivo, which is conducive to the long-term survival and functional maturation of organoids.

[0031] It is understood that the upper plate 110 has a first positioning groove 115 and a second positioning groove 116 on both sides, and the lower plate 120 has a fixing groove 129. A positioning pin 102 can be inserted between the first positioning groove 115 and the fixing groove 129, at which point the first flow channel 111 can connect to the culture chamber 126, and the chip is in the inoculation mode. When the positioning pin 102 is inserted between the second positioning groove 116 and the fixing groove 129, the second flow channel 112 can connect to the culture chamber 126, and the chip is in the culture mode. Through the cooperation of the positioning pin 102 with different positioning grooves, the operator can visually confirm the correct mode switching position, and the positioning pin 102 fixes the upper plate 110 and the lower plate 120, avoiding flow channel misalignment or leakage due to inaccurate sliding position, thereby improving the success rate and repeatability of the experiment.

[0032] The base 100 is rectangular in shape, with a rectangular observation window 105 in the center. The projection of the observation window 105 in the vertical direction completely covers the projection of all flow channels in the corresponding direction after the lower plate 120 is placed, facilitating the observation of the culture. Guide plates 103 are provided on both sides of the base 100, extending upward from the base 100 and positioned on both sides of the upper plate 110 to guide the movement of the upper plate 110.

[0033] Understandably, the two movable positioning parts 101 are mirror-symmetrical, and the main body is an "L"-shaped flat plate. The movable part 101 is pivotally connected to the base 100 and is provided with a positioning pin 102. The positioning part 101 can rotate so that the positioning pin 102 can accurately engage with the corresponding positioning grooves of the upper plate 110 and the lower plate 120, thereby realizing the mutual positioning and fixation of the upper plate 110 and the lower plate 120.

[0034] Reference Figure 11 In a second aspect of the present invention, a processing method is provided, applied to a microfluidic chip for constructing modular cell cultures as described in the above embodiments. The method includes the following steps: First, a PDMS upper plate 110 and lower plate 120 structure are fabricated using soft photolithography. Openings are made at the inlet and outlet positions of the flow channels in the upper plate 110 to form a first injection channel 113 and a second injection channel 114. The chip and piping are then subjected to high-pressure steam sterilization. Next, 0.2-10 μL of silicone oil or fluorinated oil is dropped onto the upper surface of the lower plate 120, and the upper plate 110 and lower plate 120 are slowly assembled, ensuring smooth sliding between them. This step utilizes the dual lubrication and sealing properties of the oil phase: on the one hand, it ensures that the microstructure is not damaged during relative sliding of the upper and lower plates; on the other hand, it prevents leakage of subsequently injected hydrogel precursor material from the gaps between the plates.

[0035] The assembled microfluidic chip is placed on the base 100, and the upper plate 110 is slid to the preset position to enter the cell seeding mode. The positioning pins 102 of the positioning element 101 are inserted into the first positioning groove 115 and the fixing groove 129 to fix the upper plate 110 and the lower plate 120 to each other. A pipette is used to inject a solution of hydrogel precursor material mixed with cells or organoids into the first channel 111 through the first injection channel 113 until the channel is full. The hydrogel precursor material can be Matrigel, decellularized extracellular matrix (dECM), type I collagen, type IV collagen, methacrylamide gelatin (GelMA), methacrylamide hyaluronic acid (HAMA), or alginate, etc. For alginate gel, the chip needs to be slid to the culture mode in advance, and calcium chloride solution should be introduced into the second channel 112 to gel it. Then, the calcium chloride solution is pushed out with PBS buffer and the channel is washed. Other materials can be gelled by crosslinking with temperature or ultraviolet light. Since this implementation method does not rely on 3D bioprinting, there is no need to modify or replace mainstream organoid culture materials (such as matrix gel), thus avoiding the influence of additional variables on the uncertainty of experimental results.

[0036] After gelation, the positioning element 101 is released, and the upper plate 110 is slid to the culture mode position. The chip is then re-fixed by inserting the positioning pin 102 into the second positioning groove 116 and the fixing groove 129. At this time, the second flow channel 112 is connected to the culture chamber 126. The inlet of the second flow channel 112 is connected to the pump or storage tank through the second injection channel 114, and the outlet of the second flow channel 112 is connected to the pump, waste liquid tank, or storage tank. The chip is dynamically perfused with culture medium at a set flow rate. Dynamic perfusion can simulate the shear force of fluids in vivo, promote nutrient exchange and metabolite excretion in organoids, and is conducive to the formation of larger and more functional cultures. Then, the chip and the base 100 are transferred to a cell culture incubator to begin cell culture.

[0037] After culturing for the appropriate number of days, remove the liquid perfusion system and carefully move the upper plate 110 and lower plate 120 horizontally until completely separated, exposing the cells or organoids to air. Using a pipette tip with the tip cut off, aspirate 2-40 μL of the corresponding culture medium, carefully rinse the target culture, and then gently aspirate the liquid to detach the culture from the lower plate 120 and into the pipette tip, transferring the culture to a new culture environment. Next, using pipette tips or forceps, combine cultures containing the same or different cells or organoids to form the target multicellular partition model (i.e., culture model 104) (e.g., ...). Figure 9(As shown). It is understandable that cultures of different shapes can be nested within each other or stacked along their thickness. Then, a small amount of the same type of hydrogel precursor material is added to bind them together as a whole, and the corresponding culture medium is added for further cultivation or for subsequent experiments. Since each culture has a specific macroscopic geometry (such as L-shape, T-shape, rectangle, etc.), they can be arbitrarily combined and stacked in three-dimensional space like "Tetris blocks," thus flexibly constructing a multi-cell co-culture model 104 with complex spatial heterogeneity. This modular combination method overcomes the dual bottlenecks of traditional mixed culture's inability to partition and the limitations of 3D printing materials and expensive equipment, achieving high-degree-of-freedom and highly biomimetic tissue model construction while maintaining cell viability and culture integrity.

[0038] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A microfluidic chip for constructing modular cell cultures, characterized in that, include: The upper plate is equipped with a first flow channel and a second flow channel; The lower plate is provided with multiple spaced-apart culture chambers, and each of the multiple culture chambers has a different cross-sectional shape along the horizontal direction, so as to shape cultures with different shapes. The upper plate is slidably connected to the lower plate. When the first flow channel and the culture chamber overlap, they can communicate with each other, so that inoculation can be performed into the culture chamber through the first flow channel. When the second flow channel overlaps with the culture chamber, the second flow channel can be circulated with culture medium to cultivate the culture in the culture chamber; The cultures with different shapes can be stacked together to form a culture model.

2. A microfluidic chip for constructing modular cell cultures according to claim 1, characterized in that, The lower plate is provided with a plurality of spaced-apart culture zones, each of which has a plurality of culture chambers, and the culture chambers in each culture zone have different cross-sectional shapes along the horizontal direction.

3. A microfluidic chip for constructing modular cell cultures according to claim 2, characterized in that, The first flow channel is provided with a plurality of spaced-apart connecting cavities. When the first flow channel overlaps with the plurality of culture cavities, the plurality of connecting cavities and the plurality of culture cavities are arranged alternately, and two adjacent connecting cavities can simultaneously connect to the culture cavities located between them, so that the two adjacent culture cavities can be interconnected, thereby enabling inoculation into the culture cavities through the first flow channel.

4. A microfluidic chip for constructing modular cell cultures according to claim 3, characterized in that, The culture area includes a first region and a second region. The culture chambers in both regions include a base and a protrusion that are interconnected. The protrusion extends outward from one side of the base. The protrusion of the culture chamber in the first region is connected to the end of the base, and the protrusion of the culture chamber in the second region is connected to the middle of the base.

5. A microfluidic chip for constructing modular cell cultures according to claim 4, characterized in that, The communicating cavity includes a first communicating portion and a second communicating portion. When the communicating cavity is connected to the culture cavity, a portion of the first communicating portion can overlap with a portion of the base, and a portion of the second communicating portion can overlap with a portion of the protrusion. And / or, Both ends of the first flow channel and the second flow channel are respectively provided with a first injection channel and a second injection channel. The first injection channel is used to inject cell fluid, and the second injection channel is used to inject culture medium.

6. A microfluidic chip for constructing modular cell cultures according to claim 2, characterized in that, The culture area includes a third region and a fourth region. The culture chambers in both the third and fourth regions have rectangular cross-sections. The aspect ratio of the culture chamber in the third region is not equal to that of the culture chamber in the fourth region.

7. A microfluidic chip for constructing modular cell cultures according to claim 1, characterized in that, The upper plate has a first positioning groove and a second positioning groove on both sides, and the lower plate has a fixing groove. A positioning pin can be inserted between the first positioning groove and the fixing groove so that the first flow channel can connect to the culture chamber. When the positioning pin is inserted between the second positioning groove and the fixing groove, the second flow channel can connect to the culture chamber.

8. A microfluidic chip for constructing modular cell cultures according to claim 7, characterized in that, It also includes a base, the lower plate is fixed to the base, the base is provided with a pivotally connected positioning member, and the positioning member is provided with the positioning pin.

9. A microfluidic chip for constructing modular cell cultures according to claim 8, characterized in that, The base is provided with a guide plate that extends upward from the base and is located on both sides of the upper plate to guide the movement of the upper plate.

10. A processing method applied to a microfluidic chip for constructing modular cell cultures as described in any one of claims 1 to 9, characterized in that, Includes the following steps: The upper plate, lower plate, and base are manufactured and then sterilized. Cover the lower plate with the upper plate, and install the lower plate onto the base; The upper plate is slid to connect the first flow channel to the culture chamber, and is inserted into the first positioning groove and the fixing groove through the positioning pin of the positioning member, so that the upper plate and the lower plate are fixed to each other; The solution containing the cells and precursor material is injected into the first flow channel through the first injection channel to inject into the culture chamber; Remove the positioning component to move the upper plate relative to the lower plate so that the second flow channel connects to the culture chamber, and re-fix the upper plate and the lower plate by inserting the positioning pin into the second positioning groove and the fixing groove; Connect one end of the second injection channel to the perfusion system, and inject culture medium into the second channel through the perfusion system so that the culture medium flows through the culture chamber. At the same time, connect the other end of the second injection channel to the waste liquid tank. After a preset incubation time, the upper plate and the lower plate are separated, the culture formed in the incubation chamber is removed, and the culture is placed in a new incubation environment. Multiple cultures with different shapes are stacked together to form a culture model.

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