Cavity structure adaptive to Faraday wave assembly

Abstract

The utility model relates to the technical field of cell culture, and discloses a chamber structure adaptive to Faraday wave assembly, which comprises a pore plate, N mounting grooves are formed in the top surface of the pore plate, and N is greater than or equal to 1; the N container bodies are arranged on the pore plate in a rectangular array mode or a central symmetry mode and correspondingly arranged in the N mounting grooves, a plurality of culture grooves used for cell assembly are sequentially formed in the container bodies in the longitudinal direction from the top face to the bottom face, and the widths of the culture grooves are gradually decreased. According to the device, internal cells or cell microspheres can be gathered into a sound potential well of a standing wave sound field under the action of acoustic pressure, gravity and buoyancy under the action of the sound field and are tightly arranged according to an expected pattern, and a culture solution is longitudinally stacked in a plurality of culture tanks of which the widths are gradually reduced; longitudinal shape limitation on the tissue structure is realized, and complex three-dimensional longitudinal arrangement is constructed through mutual combination of the two. Therefore, the requirement of constructing any multi-scale and complex cell structure is met. And forming a multi-layer structure.

Description

Technical Field

[0001] The utility model belongs to the field of cell culture, and in particular relates to a chamber structure adapted for Faraday wave assembly. Background Art

[0002] Acoustic bioassembly is the use of the physical principle of the interaction between the acoustic field and the assembly unit to assemble biological particles into specific tissue structures. Faraday wave acoustic assembly is a representative type of acoustic assembly. Its basic principle is to generate a surface standing wave at the gas-liquid interface and use this standing wave to drive the assembly unit to form various specific patterns. This method uses a simple device, is cost-effective, and can assemble cells in vitro in a rapid, non-destructive, and non-contact manner, thereby producing specific tissue structures. It is an ideal technology for constructing biomimetic tissues and organs in vitro. Based on Faraday wave bioassembly technology, highly biomimetic cardiac microtissues, brain-like tissues and other complex physiological structures have been successfully constructed. Existing traditional acoustic bioassembly usually uses a single-dimensional chamber for culture. When the assembly units are located at the bottom of the chamber and arranged under the drive of the acoustic field, they can usually only form a single-layer structure. Utility Model Content

[0003] In order to solve the deficiency that the assembly units are located at the bottom of the chamber and can usually only form a single-layer structure when arranged under the drive of the sound field, the present invention provides a chamber structure suitable for Faraday wave assembly.

[0004] The technical effects to be achieved by the present invention are achieved through the following technical aspects:

[0005] The utility model provides a chamber structure adapted for Faraday wave assembly, comprising:

[0006] The orifice plate has N mounting slots on the top surface, and N ≥ 1;

[0007] N container bodies are arranged in a rectangular array or a centrally symmetrical manner on the orifice plate and are correspondingly arranged in the N mounting grooves. A plurality of culture grooves for cell assembly are sequentially opened longitudinally from the top surface to the bottom surface, and the width of each culture groove is gradually reduced.

[0008] In one embodiment, a weight-reducing groove is provided on the bottom surface of the container body, so that a tower-shaped culture mold is formed between each culture groove and the weight-reducing groove.

[0009] In one embodiment, the container body is clamped in the orifice plate, the culture tank at the bottom passes through the weight loss tank, and the bottom surface of the tower-shaped culture mold is covered with a nutrient exchange membrane.

[0010] In one embodiment, the nutrient exchange membrane is configured as a polycarbonate membrane, a polyester membrane, or a collagen-coated polytetrafluoroethylene membrane.

[0011] In one embodiment, the culture tank at the bottom passes through the weight loss tank, and a transparent observation piece is embedded on the side wall of the culture tank at the bottom.

[0012] In one embodiment, the width of each of the culture tanks is set to decrease in equal intervals.

[0013] In one embodiment, a hydrophilic layer is provided at the bottom of the lowest culture tank, and a hydrophobic layer is provided at the bottom of the remaining culture tanks and on the side walls of each culture tank.

[0014] In one embodiment, the hydrophilic layer is configured as a hydrophilic coating or a hydrophilic surface, and the hydrophobic layer is configured as a hydrophobic coating or a hydrophobic surface.

[0015] In one embodiment, the cross-sectional shape of each culture tank is set to be circular or polygonal.

[0016] In one embodiment, the cross-sectional shape of the container body is set to be circular or polygonal.

[0017] In summary, the present invention has at least the following advantages:

[0018] The Faraday wave-adapted chamber structure provided by the present invention not only utilizes the acoustic field to drive the patterned arrangement of the culture fluid in each layer of culture tanks, but also, by providing multiple culture tanks of decreasing width, allows the culture fluid to be stacked vertically within the multiple culture tanks, thereby constraining the longitudinal shape of the tissue structure. In this way, the two can be combined to construct complex three-dimensional longitudinal arrangements, thereby achieving the goal of constructing multi-scale, complex, and arbitrary cell structures, i.e., forming a multi-layered structure. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 1 is a schematic diagram of a three-dimensional structure of a container body of a culture tank having a circular cross-section according to an embodiment of the present application;

[0020] Figure 2 1 is another schematic diagram of the three-dimensional structure of the container body of the culture tank of the embodiment of the present application, wherein the cross-section thereof is circular;

[0021] Figure 3 1 is a schematic cross-sectional structural diagram of a culture tank according to an embodiment of the present application, wherein the cross-sectional shape of the container body is circular;

[0022] Figure 4 3D schematic diagram of a chamber structure of a culture tank having a circular cross-section and adapted for Faraday wave assembly according to an embodiment of the present application;

[0023] Figure 5 1 is a schematic diagram of a three-dimensional structure of a container body of a culture tank according to an embodiment of the present application, wherein the cross-sectional shape of the container body is an equilateral triangle;

[0024] Figure 6 1 is another schematic diagram of the three-dimensional structure of the container body of the culture tank of an embodiment of the present application, wherein the cross-sectional shape of the container body is an equilateral triangle;

[0025] Figure 7 This is a partial cross-sectional structural schematic diagram of a chamber structure adapted for Faraday wave assembly with a snap-fit ​​portion and provided with a nutrient exchange membrane according to an embodiment of the present application;

[0026] Figure 8 This is a partial cross-sectional structural schematic diagram of a chamber structure adapted for Faraday wave assembly with a clamping portion and a transparent observation piece according to an embodiment of the present application;

[0027] Figure 9 1 is a schematic diagram of a three-dimensional structure of a chamber structure having a clamping portion adapted for Faraday wave assembly according to an embodiment of the present application;

[0028] 10. Chamber structure adapted for Faraday wave assembly; 100. Orifice plate; 110. Mounting slot; 200. Container body; 210. Culture tank; 211. Hydrophilic layer; 212. Hydrophobic layer; 220. Weight reduction tank; 230. Tower-shaped culture mold; 240. Nutrient exchange membrane; 250. Transparent observation piece. DETAILED DESCRIPTION

[0029] To make the purpose, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. The described embodiments are only part of the embodiments of the present invention, not all of the embodiments.

[0030] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but rather merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0031] Example 1:

[0032] It is understandable that the culture medium is a liquid, solidifiable culture medium containing living cells. The living cells used can be stem cells such as embryonic stem cells, mesenchymal stem cells, etc., or one or more mixtures of primary cells of tissue organs such as liver and kidney, or cell microspheres or cell carrier microparticles. The diameter of the living cells can be between a few μm and several thousand μm. The sound field in the sound field assembly can be a Faraday wave or a standing wave field at the gas-liquid interface. The orifice plate 100 can be a transparent orifice plate 100, for example: made of transparent materials such as PS (polystyrene), which is convenient for observation and has certain heat resistance and good biocompatibility. The material of the container body 200 can be set to PS, PC (polycarbonate) or PMMA (polymethyl methacrylate).

[0033] like Figures 1 to 9 As shown, the present invention provides a chamber structure 10 adapted for Faraday wave assembly, comprising: a well plate 100, with N mounting grooves 110 provided on the top surface, and N ≥ 1; N container bodies 200, arranged on the well plate 100 in a rectangular array or in a centrally symmetrical manner, and correspondingly arranged in the N mounting grooves 110, and a plurality of culture grooves 210 for cell assembly are provided longitudinally from the top surface to the bottom surface, and the width of each culture groove 210 is set to decrease successively.

[0034] Specifically, N is a positive integer greater than or equal to 1, the cross-section of the orifice plate 100 is set to be rectangular, and N mounting grooves 110 for mounting the container bodies 200 are provided on the top surface of the orifice plate 100, and the shape of the mounting grooves 110 matches the shape of the container bodies 200, and the number of the mounting grooves 110 matches the number of the container bodies 200, so that the container bodies 200 can be mounted on the orifice plate 100 through the mounting grooves 110. N container bodies 200 are arranged in a rectangular array on the orifice plate 100, or N container bodies 200 are arranged in a center-symmetrical manner on the orifice plate 100, and the container bodies 200 are arranged axially symmetrically, so that each container body 200 has good radiation and symmetry, better meets the mechanical response, and ensures the uniformity of assembly. Reference Figures 1 to 6, the cross-sectional shape of the container body 200 can be set to a circle, wherein the circumscribed circle diameter of the container body 200 is 0.5cm to 20cm. For example, the circumscribed circle diameter of the container body 200 is 0.5cm. For another example, the circumscribed circle diameter of the container body 200 is 10.25cm. For another example, the circumscribed circle diameter of the container body 200 is 20cm. It can be understood that the cross-sectional shape of the container body 200 can also be set to other shapes, such as positive polymorphic shapes, which are not described in detail in this embodiment. There are multiple culture tanks 210 on the top surface of the container body 200, and the width of each culture tank 210 is gradually reduced from top to bottom, so that the longitudinal cross-sectional shape combination of each culture tank 210 is arranged in a tower shape. And with reference to Figures 1 to 4 The cross-sectional shape of each culture tank 210 can be set to a circular shape. It can be understood that the cross-sectional shape of each culture tank 210 can also be set to other shapes. Figure 5 and Figure 6 , such as regular polygons, are not described in detail in this embodiment. The regular polygons may be regular triangles, squares, regular pentagons, regular hexagons, or regular octagons.

[0035] When performing acoustic field assembly, a Faraday wave with a driving frequency of 1 to 1000 Hz is used. For example, a Faraday wave of 1 Hz is used; for another example, a Faraday wave of 500 Hz is used; for another example, a Faraday wave of 1000 Hz is used. This range is conducive to the Faraday wave acoustic field driving the cell-containing assembly unit, and is also conducive to the relative stability of the gas-liquid-solid phase. When the culture fluid is added to the culture tank 210, it is added in batches. The first layer of culture fluid needs to be solidified after addition, and the second layer of culture fluid is added after solidification. This process is repeated until the desired pattern is stacked vertically, and the top culture tank 210 is empty. The barrier effect of the top culture tank 210 can prevent the culture fluid in the lower culture tank 210 from overflowing, so that the culture fluid can be assembled in each culture tank 210 below in sequence without overflowing. The shape of the container body 200 creates favorable culture conditions for cell assembly. When performing acoustic assembly to construct spatially specific three-dimensional in vitro biological tissues, the vertical stacking of the assembly chambers allows each layer to construct a corresponding pattern according to design requirements, ultimately forming a complex tissue structure in the vertical and horizontal directions. In other words, in addition to using the acoustic field to drive the culture fluid in each layer of the culture tank 210 to form a pattern, the container body 200 can also achieve longitudinal shape constraints on the tissue structure through the vertical stacking of the culture fluid within the culture tank 210. The two can be combined to construct a complex three-dimensional longitudinal arrangement, thereby meeting the needs of constructing multi-scale, complex, and arbitrary cell structures.

[0036] In order to facilitate the preparation of the container body 200, as Figure 3As shown, in one embodiment, the width of each culture tank 210 is set to decrease in equal intervals.

[0037] Specifically, the thickness of each culture tank 210 is equal, and the thickness of each culture tank 210 can be 0.2 mm to 10 mm. For example, the thickness of each culture tank 210 can be 0.2 mm; for another example, the thickness of each culture tank 210 can be 5.1 mm; for another example, the thickness of each culture tank 210 can be 10 mm. The upper culture tank 210 can extend 0.5 cm to 2 cm farther than the lower culture tank 210. For example, the upper culture tank 210 can extend 0.5 cm farther than the lower culture tank 210; for another example, the upper culture tank 210 can extend 1.25 cm farther than the lower culture tank 210; for another example, the upper culture tank 210 can extend 2 cm farther than the lower culture tank 210. The same process is repeated downward, thereby achieving pattern superposition. For example, under single wavelength conditions, a pattern of multiple layers of diverging, petal-shaped, and concentric ring patterns can be obtained.

[0038] It is understood that the number of mounting slots 110 is designed according to the number of container bodies 200. In this embodiment, N=6, that is, the orifice plate 100 is provided with six mounting slots 110, which is a six-orifice plate 100. It can also be provided as a twelve-orifice plate 100, a twenty-four-orifice plate 100, a forty-eight-orifice plate 100, a ninety-six-orifice plate 100, and the like.

[0039] Example 2:

[0040] The difference between this embodiment and embodiment 1 is that this embodiment further optimizes the structure of the chamber structure 10 adapted for Faraday wave assembly of the present invention. Figure 3 As shown, a hydrophilic layer 211 is provided at the bottom of the bottommost culture tank 210 , and a hydrophobic layer 212 is provided at the bottom of the remaining culture tanks 210 and on the side walls of each culture tank 210 .

[0041] Specifically, the bottom of the bottommost culture tank 210 is provided with a hydrophilic layer 211, which facilitates the filling of the culture solution and subsequent assembly of the culture solution. Furthermore, the bottoms of the remaining culture tanks 210 and the sidewalls of each culture tank 210 are provided with a hydrophobic layer 212, which facilitates the assembly of the culture solution. Furthermore, the combination of the hydrophilic layer 211 and the hydrophobic layer 212 improves the assembly effect.

[0042] Furthermore, the hydrophilic layer 211 is configured as a hydrophilic coating or a hydrophilic surface, and the hydrophobic layer 212 is configured as a hydrophobic coating or a hydrophobic surface.

[0043] Specifically, the hydrophilic layer 211 is configured as a hydrophilic coating. The hydrophilic coating may be made of materials such as polyvinyl alcohol, PHEMA (poly(2-hydroxyethyl methacrylate)), PEG (polyethylene glycol), zwitterions, amino acids, or silicone gel. The hydrophobic layer 212 is configured as a hydrophobic coating. The hydrophobic coating may be made of materials such as PDMS (polydimethylsiloxane), PMMA, PTFE (polytetrafluoroethylene), or a silane coupling agent. Both the hydrophilic and hydrophobic coatings can be fabricated using conventional methods, such as sol-gel methods, thermal initiation methods, photocatalytic methods, coating methods, and transfer methods.

[0044] It is understandable that the hydrophilic layer 211 and the hydrophobic layer 212 may also be made in other ways.

[0045] In one embodiment, the hydrophilic layer 211 is configured as a hydrophilic material surface. For example, the hydrophilic layer 211 can be prepared by thinly coating PEGDA [polyethylene glycol diacrylate] on the inner wall of the well plate 100 and then irradiating the well plate 100 with light at a wavelength of 405 nm for 10 minutes. For another example, the hydrophilic layer 211 can be prepared by thinly coating zwitterions and polyvinyl alcohol on the inner wall of the well plate 100 and then drying at 60° C. for 6 hours. The hydrophobic layer 212 is configured as a hydrophobic material surface. For example, the hydrophobic layer 212 can be prepared by thinly coating PDMS on the inner wall of the well plate 100 and then drying at 60° C. for 10 hours. For another example, the hydrophobic layer 212 can be prepared by thinly coating a silane coupling agent on the inner wall of the well plate 100 and then drying at 60° C. for 5 hours.

[0046] In another embodiment, the hydrophilic layer 211 is configured as a hydrophilic coating, and the hydrophobic layer 212 is configured as a hydrophobic surface.

[0047] In yet another embodiment, the hydrophilic layer 211 is configured as a hydrophilic surface, and the hydrophobic layer 212 is configured as a hydrophobic coating.

[0048] In addition, the hydrophilic layer 211 and the hydrophobic layer 212 may be manufactured in other ways known to those skilled in the art, which will not be described here in detail.

[0049] Example 3:

[0050] The difference between this embodiment and embodiment 1 is that this embodiment further optimizes the structure of the chamber structure 10 adapted for Faraday wave assembly of the present invention. Figure 3As shown, a weight-reducing groove 220 is provided on the bottom surface of the container body 200 so that a tower-shaped culture mold 230 is formed between each culture groove 210 and the weight-reducing groove 220 .

[0051] Specifically, the presence of a weight-reducing groove 220 on the bottom surface of the container body 200 reduces weight, minimizes material loss, and effectively lowers production costs. When the container body 200 is placed in the mounting groove 110, the culture medium added to the mounting groove 110 can flow into the weight-reducing groove 220, preventing overflow of the culture medium from the mounting groove 110 due to the larger volume of the container body 200. This forms a tower-shaped culture mold 230.

[0052] Example 4:

[0053] The difference between this embodiment and embodiment 3 is that this embodiment further optimizes the structure of the chamber structure 10 adapted for Faraday wave assembly of the present invention. Figure 7 and Figure 9 As shown, the bottommost culture tank 210 is set through the weight loss tank 220, and the bottom end of the tower-shaped culture mold 230 is covered with a nutrient exchange membrane 240.

[0054] Specifically, the container body 200 is provided with protruding clamping parts on both sides, so that the container body 200 is clamped on the orifice plate 100 through the two clamping parts. At this time, the bottom surface of the container body 200 is spaced apart from the bottom of the mounting groove 110. By connecting the culture groove 210 at the bottom with the weight loss groove 220, it is convenient for each culture groove 210 to interact with the weight loss groove 220. The nutrient exchange membrane 240 is provided to block the bottom end of the culture groove 210 at the bottom. The nutrient exchange membrane 240 is a polycarbonate membrane, and the nutrient exchange membrane 240 is spaced apart from the bottom of the mounting groove 110. The separation effect of the nutrient exchange membrane 240 can make the chamber formed by each culture groove 210 an upper chamber, and the weight loss groove 220 and the mounting groove 110 a lower chamber. The culture medium can be added to the upper chamber to perform cell assembly. In this way, when in use, the container body 200 containing culture medium is placed in the mounting tank 110, and culture medium is added to the mounting tank 110. The culture medium in the mounting tank 110 can overflow into the weight loss tank 220 and contact the nutrient exchange membrane 240. At this time, the nutrient exchange membrane 240 allows cells to secrete and absorb nutrient molecules from the bottom and top surfaces, thereby allowing the cells in the upper chamber to metabolize in a more natural way. Furthermore, the container body 200 can be moved to facilitate transfer of culture.

[0055] In another embodiment, the nutrient exchange membrane 240 is configured as a polyester film.

[0056] In yet another embodiment, the nutrient exchange membrane 240 is configured as a collagen-coated polytetrafluoroethylene membrane.

[0057] Example 5:

[0058] The difference between this embodiment and embodiment 3 is that this embodiment further optimizes the structure of the chamber structure 10 adapted for Faraday wave assembly of the present invention. Figure 8 and Figure 9 As shown, the culture tank 210 at the bottom is arranged to pass through the weight loss tank 220, and a transparent observation piece 250 is embedded on the side wall of the culture tank 210 at the bottom.

[0059] Specifically, the container body 200 has protruding snap-fitting portions on both sides, allowing the container body 200 to be secured to the well plate 100 via the two snap-fitting portions. At this point, the bottom surface of the container body 200 is spaced apart from the bottom of the mounting groove 110. By connecting the bottommost culture groove 210 with the weight-reducing groove 220, fluid exchange between each culture groove 210 and the weight-reducing groove 220 is facilitated. A transparent observation plate 250 tightly fits into the sidewalls of the bottommost culture groove 210, with its bottom surface flush with the bottom surface of the tower-shaped culture mold 230. The transparent observation plate 250 is made of high-transparency borosilicate glass, which has high optical properties and ensures optimal bottom flatness. Borosilicate glass can have a diameter of 20 mm and a thickness of 0.17 mm ± 0.02 mm. It is also heat- and endotoxin-free. This facilitates direct culture of tissue and organ analogs in a culture dish and allows operators to easily observe the assembly process within the container body 200.

[0060] The material of the transparent observation piece 250 may also be other optically transparent materials, including: quartz, COC (copolymers of cycloolefin), COP (Cyclo Olefin Polymer), PDMS, PMMA, PC, PS, resin agarose or one or more of them used in combination.

[0061] Example 6:

[0062] Based on the above embodiments, this embodiment provides an assembly method of a chamber structure 10 adapted for Faraday wave assembly, including:

[0063] S1: Pour the culture solution into the culture tank 210 at the bottom layer and perform the acoustic field assembly and curing operation to obtain the first layer of gel;

[0064] S2: Continue pouring the culture solution into the penultimate culture tank 210, so that the second layer of culture solution is assembled and solidified on the first layer of gel by acoustic field, thereby obtaining a second layer of gel;

[0065] S3: Repeat this process until all required culture tanks 210 are filled with gel, so as to assemble a three-dimensional structure in the container body 200.

[0066] Specifically, in the process of assembling the acoustic field, the culture fluid is added to the culture tank 210 and the volume of the culture fluid added is controlled, thereby controlling the volume of the culture fluid to each layer of the culture tank 210. Because the acoustic field is a Faraday wave or a standing wave field at the gas-liquid interface, it can induce hydraulic instability by periodically vibrating the liquid layer vertically. When the vibration acceleration reaches a certain threshold, it generates a liquid surface standing wave. Therefore, when the Faraday wave or the standing wave field at the gas-liquid interface is turned on, the culture fluid can vibrate accordingly, and the cells or cell microspheres therein are gathered into the acoustic potential well of the standing wave acoustic field under the action of acoustic pressure, gravity, and buoyancy, and are tightly arranged according to the expected pattern. Through operations such as ultraviolet light irradiation or cooling, the culture fluid is solidified into a gel, and its surface remains smooth and flat. After the layer of culture fluid is completely solidified and has a certain strength, the next layer of culture fluid is added, and the above steps are repeated to finally complete the assembly operation. The multi-layer stacking can be effectively completed, and a dense and uniform interface is formed between the layers, which is conducive to the exchange of nutrients and oxygen between the cells in the chamber formed by each culture tank. After multi-layer acoustic field assembly, the resulting solidified gel is cultured under appropriate conditions, allowing cells to grow into tissues or organs with biomimetic structures and activities. In addition to using the acoustic field to drive the patterned arrangement of each layer of culture fluid, the vertical stacking of the culture fluids also constrains the shape of the tissue structure, thereby constructing complex three-dimensional shapes.

[0067] Example 7

[0068] This embodiment, based on the fourth embodiment, provides an assembly method of a chamber structure 10 adapted for Faraday wave assembly, including:

[0069] S1: The culture medium is set to 180 μL of GelMA (methacryloyl) hydrogel mixed with HUVEC (Human Umbilical Vein Endothelial Cells). The culture medium is poured into the culture tank 210 at the bottom layer, and the signal frequency of the function signal generator is adjusted to 110 Hz and the amplitude to 340 mVpp. Then, an acoustic field assembly and curing operation is performed to obtain an assembly mode in which living cells are tightly connected to form a gel pattern, thereby assembling a cured single layer of gel in the container body 200.

[0070] S2: GM growth medium is added to the container body 200, and the volume of the GM growth medium does not cover the solidified gel, and the container body is placed in a 5% CO2, 37°C incubator for 24 hours to obtain a three-dimensional gel.

[0071] Among them, the main reagents used are as follows:

[0072] GM growth medium: ECM (Endothelial Cell Medium) special culture medium;

[0073] GelMA hydrogel: An edible, photosensitive liquid gel. GM and GelMA solids are mixed in a suitable ratio and dissolved in a 70°C water bath until no precipitation or dense bubbles remain. The solution is then sterilized by filtration using a 0.22 µm filter. Before use, the solution must be warmed in a 37°C metal bath.

[0074] Specifically, when the Faraday wave or the standing wave field at the gas-liquid interface is turned on, the culture medium can vibrate accordingly, and the cells therein gather in the acoustic potential well of the standing wave field under the action of acoustic pressure, gravity, and buoyancy, and are tightly arranged according to the expected pattern. The GelMA hydrogel is cross-linked and solidified by ultraviolet light, and its surface remains smooth and flat, and finally the assembly operation is completed. The single-layer gel prepared by this method is observed under a microscope, and it can be seen that the cells are almost undamaged and the viability is considerable; microscopic observation of the aggregation degree of the local area of ​​the single-layer gel shows that the cells in the single-layer gel are very close.

[0075] Example 8

[0076] This embodiment is a further implementation of Example 6. In this embodiment, the Faraday wave driving frequency used is 50-200 Hz, and the culture medium is GelMA hydrogel mixed with HUVEC to assemble the muscle cell ball unit. The cross-sectional shape of each culture tank 210 is circular. After the acoustic assembly is completed, the assembled three-dimensional structure is transferred to a culture dish for culture.

[0077] Specifically, a hydrogel cell suspension is assembled from bottom to top at a specific frequency and amplitude. After patterning, the hydrogel is solidified to create patterns of concentric circles, hexagonal petals, concentric circles, and octagonal petals, respectively. After solidification, the hydrogel is added to a suitable culture medium and cultured conventionally. Cells can be observed sprouting and growing, forming muscle fiber bundles with distinct patterns, ultimately forming a block of artificial meat.

[0078] Example 9

[0079] This embodiment is a further implementation of Example 6. In this embodiment, the driving frequency of the Faraday wave used is 50-200 Hz, and the culture medium is GelMA hydrogel mixed with HUVEC to assemble the muscle cell ball unit. The cross-sectional shape of each culture tank 210 is square. After the acoustic assembly is completed, the assembled three-dimensional structure is transferred to a culture dish for culture.

[0080] Specifically, the hydrogel cell suspension is assembled from bottom to top at a certain frequency and amplitude. After patterning, the hydrogel is solidified to obtain strip, short strip, circular flower and grid patterns. After proper culture, the cells begin to grow in the hydrogel. The cells are connected to each other and have a certain arrangement order, which increases the material exchange and signal transmission between cells and provides a good environment for cell differentiation and fusion.

[0081] Example 10

[0082] This embodiment is a further implementation of Example 6. In this embodiment, the cross-sectional shape of each culture tank 210 is square. The container body 200 is first fixed to the vibrator so that the culture tank 210 at the bottom layer can be affected by the vibrator. The culture medium is GelMA hydrogel mixed with HUVEC. Then, the culture medium is added through the assembly method of Example 6 for culture. After the last layer is assembled and fixed, the assembled three-dimensional structure is separated from the chamber for conventional culture.

[0083] In this way, a completely peeled hydrogel cell culture can be obtained, and the hydrogel cell culture has a beautiful morphology.

[0084] In this utility model, unless otherwise specified or limited, the terms "installed," "connected," "connect," "fixed," etc. should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integration; mechanical connection, electrical connection; direct connection, indirect connection through an intermediate medium, internal communication between two components, or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in this utility model based on specific circumstances.

[0085] In the description of this utility model, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer" and the like, indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, or are the orientations or positional relationships in which the utility model product is typically placed when in use. These terms are intended solely to facilitate the description of this utility model and to simplify the description, and are not intended to indicate or imply that the device or component referred to must have a specific orientation, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limitations on this utility model. Furthermore, the terms "first," "second," and "third," etc., are used solely to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0086] Furthermore, terms such as "horizontal," "vertical," and "overhanging" do not necessarily imply that a component must be absolutely horizontal or overhanging, but rather that it can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal than "vertical," and does not mean that the structure must be completely horizontal, but rather that it can be slightly tilted.

[0087] In the present invention, unless otherwise expressly specified or limited, a first feature being above or below a second feature may include the first and second features being in direct contact, or may include the first and second features being in contact not directly but through another feature between them. Furthermore, a first feature being above, above, and above the second feature includes the first feature being directly above and obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being below, below, and below the second feature includes the first feature being directly below and obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0088] Although the present invention has been described with reference to the above specific embodiments, it is obvious that those skilled in the art can make many substitutions, modifications and variations based on the above content. Therefore, all such substitutions, modifications and variations are included within the spirit and scope of the appended claims.

Claims

1. A chamber structure adapted for Faraday wave assembly, characterized in that: include: The orifice plate (100) has N mounting grooves (110) on its top surface, and N is greater than or equal to 1; N container bodies (200) are arranged on the orifice plate (100) in a rectangular array or in a centrally symmetrical manner, and are correspondingly arranged in the N mounting grooves (110). A plurality of culture grooves (210) for cell assembly are sequentially provided longitudinally from the top surface to the bottom surface, and the width of each culture groove (210) is successively decreased.

2. The chamber structure adapted for Faraday wave assembly according to claim 1, characterized in that: A weight-reducing groove (220) is provided on the bottom surface of the container body (200), so that a tower-shaped culture mold (230) is formed between each culture groove (210) and the weight-reducing groove (220).

3. The chamber structure adapted for Faraday wave assembly according to claim 2, characterized in that: The container body (200) is stuck in the orifice plate (100), the bottom culture tank (210) is set through the weight loss tank (220), and the bottom surface of the tower-shaped culture mold (230) is covered with a nutrient exchange membrane (240).

4. The chamber structure adapted for Faraday wave assembly according to claim 3, characterized in that: The nutrient exchange membrane (240) is configured as a polycarbonate membrane, a polyester membrane or a collagen-coated polytetrafluoroethylene membrane.

5. The chamber structure adapted for Faraday wave assembly according to claim 2, characterized in that: The bottommost culture tank (210) is arranged to pass through the weight loss tank (220), and a transparent observation piece (250) is embedded on the side wall of the bottommost culture tank (210).

6. The chamber structure adapted for Faraday wave assembly according to claim 1, characterized in that: The width of each of the culture tanks (210) is set to decrease in equidistant order.

7. The chamber structure adapted for Faraday wave assembly according to claim 1, characterized in that: The bottom of the culture tank (210) at the bottom is provided with a hydrophilic layer (211), and the bottoms of the remaining culture tanks (210) and the side walls of each culture tank (210) are provided with a hydrophobic layer (212).

8. The chamber structure adapted for Faraday wave assembly according to claim 7, characterized in that: The hydrophilic layer (211) is configured as a hydrophilic coating or a hydrophilic surface, and the hydrophobic layer (212) is configured as a hydrophobic coating or a hydrophobic surface.

9. The chamber structure adapted for Faraday wave assembly according to claim 1, characterized in that: The cross-sectional shape of each culture tank (210) is set to be circular or polygonal.

10. The chamber structure adapted for Faraday wave assembly according to claim 1, characterized in that: The cross-sectional shape of the container body (200) is set to be circular or polygonal.

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