A cell culture chip

Abstract

The embodiment of the application discloses a cell culture chip, which comprises a main body and a partitioning piece, a first channel and a second channel are formed between the partitioning piece and the main body, the partitioning piece comprises a communication section, the communication section comprises a first protrusion and a second protrusion extending along a first direction, the first protrusion and the second protrusion are spaced apart along a second direction, the side walls of the first protrusion and the second protrusion, which are away from each other along the second direction, are connected with the main body, a communication port for communicating the first channel and the second channel is formed between the first protrusion and the second protrusion, and the second direction is arranged at an angle with the first direction. The cell culture chip can form an organ-blood vessel interface between the first channel and the second channel through a gas-liquid interface formed at the communication port, exchange nutrients, gas and metabolites, and realize the culture of vascular endothelial cells and cells corresponding to organs in the first channel and the second channel respectively, without the need of additionally arranging a porous membrane, and the structure is simple and the operation is convenient.

Description

[0001] This application claims priority to Chinese Patent Application No. 2022109109926, filed on July 29, 2022, entitled “A Cell Culture Chip”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to the field of cell culture technology, and more particularly to a cell culture chip. Background Technology

[0003] Cell culture refers to the process of taking tissues from an organism and incubating them under specific in vitro conditions that simulate the physiological environment in vivo, allowing them to survive and multiply. Organ-on-a-chip technology is a technique for three-dimensional in vitro cell culture. By constructing microchannels, microreaction chambers, and other functional components, it precisely manipulates the components within the chip, such as cells, fluids, gases, and the extracellular microenvironment, thereby generating biofunctional human microtissues and microorganisms. Organ-on-a-chip technology boasts advantages such as integration, low consumption, high throughput, high simulation accuracy, and rapid analysis, and has broad application prospects in fields such as new drug development, disease models, personalized medicine, and aerospace medicine. In 2016, organ-on-a-chip technology was listed as one of the "Top Ten Emerging Technologies" by the World Economic Forum in Davos.

[0004] The human body transports nutrients and gases to various parts of the body via blood vessels, and exchanges nutrients, gases, and metabolites through the organ-blood vessel interface. To simulate this structure within the human body, many organ-on-a-chip (O-chip) studies have employed porous membranes to construct such interfaces. Vascular endothelial cells and cells corresponding to the organ are cultured on either side of the membrane to simulate the cell arrangement and interface function of the organ-blood vessel interface. However, adding porous membranes places high demands on the membrane's properties and biocompatibility. The presence of the membrane can also affect the free interaction and migration of cells between channels. Furthermore, the integration of porous membranes into the chip is relatively complex. Summary of the Invention

[0005] This application provides a cell culture chip to address the problems in related technologies where porous membranes are used to simulate the cell arrangement and interface function of organ-blood vessel interfaces. The presence of porous membranes can affect the free interaction and migration of cells between channels, and the complex binding methods of porous membranes in the chip.

[0006] This application provides a cell culture chip, including:

[0007] main body;

[0008] A partition member, located within and connected to the main body, has a first sidewall and a second sidewall facing away from each other. A first channel is formed between the partition member and the main body on the side where the first sidewall of the partition member is located, and a second channel is formed on the side where the second sidewall of the partition member is located. The first channel is used for fluid to flow in a first direction, and the second channel is used for fluid to flow in the first direction. The partition member includes a connecting section, which includes a first protrusion and a second protrusion extending in the first direction. The first protrusion and the second protrusion are spaced apart in a second direction. The sidewalls of the first protrusion and the second protrusion facing away from each other in the second direction are both connected to the main body. A connecting opening is formed between the first protrusion and the second protrusion, connecting the first channel and the second channel. The second direction is set at an angle to the first direction.

[0009] In some embodiments, the first protrusion has a first sub-sidewall and a second sub-sidewall extending along the first direction and facing away from each other, and the second protrusion has a third sub-sidewall and a fourth sub-sidewall extending along the first direction and facing away from each other, wherein the first sidewall includes the first sub-sidewall and the third sub-sidewall, and the second sidewall includes the second sub-sidewall and the fourth sub-sidewall.

[0010] The second sub-sidewall and the fourth sub-sidewall are located in the same plane; or, at least part of the second sub-sidewall is located between the first sub-sidewall and the fourth sub-sidewall.

[0011] In some embodiments, the second sub-sidewall and the fourth sub-sidewall are located in the same plane, and the first sub-sidewall and the third sub-sidewall are located in the same plane.

[0012] In some embodiments, at least a portion of the second sub-sidewall is located between the first sub-sidewall and the fourth sub-sidewall, and at least a portion of the third sub-sidewall is located between the second sub-sidewall and the fourth sub-sidewall.

[0013] In some embodiments, the second protrusion includes a central portion distributed along the first direction and an end portion connecting the central portion, the central portion being located on the side of the second sub-sidewall opposite to the first sub-sidewall, and the end portion extending in a direction bent toward the first sub-sidewall; or

[0014] The second protrusion extends in a straight line.

[0015] In some embodiments, the first protrusion is a ridge extending along the first direction; and / or, the second protrusion is a ridge extending along the first direction.

[0016] In some embodiments, the partition further includes:

[0017] Two blocking sections are distributed along the first direction on opposite sides of the connecting section, the blocking sections separating the first channel from the second channel so that the first channel and the second channel are connected only through a connecting port.

[0018] In some embodiments, the barrier segment has a fifth sub-sidewall and a sixth sub-sidewall extending along the first direction and facing away from each other, the first sidewall including the fifth sub-sidewall and the second sidewall including the sixth sub-sidewall.

[0019] In the direction from one end of the blocking segment to the other end of the connecting segment, the distance between the fifth sub-sidewall and the sixth sub-sidewall gradually increases.

[0020] In some embodiments, the angle between the fifth sub-sidewall and the sixth sub-sidewall is greater than or equal to 45° and less than or equal to 170°.

[0021] In some embodiments, the body includes:

[0022] First plate;

[0023] The second plate is stacked on top of the first plate. The partition is located between the first plate and the second plate and forms the first channel and the second channel with the first plate and the second plate. The first protrusion is connected to the first plate, and the second protrusion is connected to the second plate. The second plate also has a first through hole communicating with the first channel, a second through hole communicating with the first channel, a third through hole communicating with the second channel, and a fourth through hole communicating with the second channel.

[0024] In some embodiments, the body further includes:

[0025] The third plate is stacked on the side of the second plate away from the first plate. The third plate has a first inlet communicating with the first through hole, a first outlet communicating with the second through hole, a second inlet communicating with the third through hole, and a second outlet communicating with the fourth through hole.

[0026] In some embodiments, the cell culture chip includes two partitions, the first sidewalls of the two partitions are disposed opposite each other and a first channel is formed between the two first sidewalls, the second sidewalls of the two partitions are disposed opposite each other, and a second channel is formed on one side of the second sidewall of each partition.

[0027] The cell culture chip of this application has a first and second protrusion forming an edge structure in the partition. The presence of the edge structure changes the spreading direction of the fluid in the first channel, restricting the spread of the fluid on the solid surface and preventing the fluid in the first channel from diffusing and overflowing into the second channel through the connecting port. The fluid in the first channel is confined within the first channel, and a gas-liquid interface is formed at the connecting port to constitute an organ-blood vessel interface between the first and second channels, facilitating the exchange of nutrients, gases, and metabolites. This allows for the separate culture of vascular endothelial cells and corresponding organ cells in the first and second channels, respectively, without the need for a separate porous membrane. The structure is simple and easy to operate. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0029] Figure 1 This is a three-dimensional structural schematic diagram of a cell culture chip provided in one embodiment of this application;

[0030] Figure 2 yes Figure 1 The diagram shown is a three-dimensional exploded view of a cell culture chip.

[0031] Figure 3 yes Figure 1 A three-dimensional cross-sectional schematic diagram of the cell culture chip is shown.

[0032] Figure 4 yes Figure 1 The diagram shown illustrates the effect of adding fluid to the first channel of the cell culture chip.

[0033] Figure 5 yes Figure 1 The diagram shows a three-dimensional structure of the first plate in the cell culture chip.

[0034] Figure 6 This is a cross-sectional schematic diagram of a cell culture chip provided in another embodiment of this application, wherein the partition includes a first protrusion and a second protrusion disposed along a second direction;

[0035] Figure 7 yes Figure 6 A schematic diagram illustrating the simulation effect of the cell culture chip is shown.

[0036] Figure 8 yes Figure 6In a control group, the cell culture chip's partitions consisted of only a single protrusion positioned along the second direction.

[0037] Figure 9 yes Figure 8 A schematic diagram illustrating the simulation effect of the cell culture chip is shown.

[0038] Figure 10 This is a partial three-dimensional structural diagram of the first plate in a cell culture chip provided in another embodiment of this application;

[0039] Figure 11 yes Figure 10 The diagram shown illustrates the effect of adding fluid to the first channel of the cell culture chip.

[0040] Figure 12 yes Figure 11 A schematic diagram illustrating the effect of adding fluid into the first channel as an alternative solution;

[0041] Figure 13 This is a partial three-dimensional structural diagram of the first plate in a cell culture chip provided in another embodiment of this application;

[0042] Figure 14 yes Figure 6 In another control group, the cell culture chip partition includes a first protrusion and a second protrusion disposed along the second direction, and at least a portion of the fourth sub-sidewall of the second protrusion is located on the side of the second sub-sidewall of the first protrusion away from the first sub-sidewall of the first protrusion.

[0043] Figure 15 yes Figure 14 The diagram shows a simulation of the cell culture chip.

[0044] Explanation of reference numerals in the attached drawings: 100, cell culture chip; 110, main body; 111, first plate; 112, second plate; 1121, first through hole; 1122, second through hole; 1123, third through hole; 1124, fourth through hole; 113, third plate; 1131, first inlet; 1132, first outlet; 1133, second inlet; 1134, second outlet; 120, partition; 121, first sidewall; 122, second sidewall; 123, connecting section; 1231, first protrusion; 123 2. Second protrusion; 1233. First sub-sidewall; 1234. Second sub-sidewall; 1235. Third sub-sidewall; 1236. Fourth sub-sidewall; 1237. Middle section; 1238. End section; 124. Blocking section; 1241. Fifth sub-sidewall; 1242. Sixth sub-sidewall; 130. First channel; 131. Third sidewall; 132. Fourth sidewall; 140. Second channel; 141. Fifth sidewall; 142. Sixth sidewall; 150. Connecting port; x. First direction; y. Second direction; z. Third direction. Detailed Implementation

[0045] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings.

[0046] In the following description, when referring to the accompanying drawings, the same numbers in different drawings denote the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with this application. Rather, they are merely examples of apparatuses and methods consistent with some aspects of this application as detailed in the appended claims.

[0047] Example 1

[0048] Please see Figures 1 to 5 This application provides a cell culture chip 100, which includes a main body 110 and a partition 120.

[0049] Combination Figure 1 and Figure 2 The partition 120 is located within and connected to the main body 110. The partition 120 has a first sidewall 121 and a second sidewall 122 facing away from each other. A first channel 130 is formed between the partition 120 and the main body 110 on the side where the first sidewall 121 is located, and a second channel 140 is formed on the side where the second sidewall 122 is located. The first channel 130 is used for fluid flow along a first direction x, and the second channel 140 is used for fluid flow along the first direction x. Specifically, the first channel 130 and the second channel 140 can be used to transport at least one of the following components: cells, fluids, gases, and extracellular microenvironments.

[0050] It should be noted that the first channel 130 can be used to allow fluid to flow along the first direction x in the following ways: the first channel 130 extends along the first direction x, thereby allowing fluid within the first channel 130 to flow along the first direction x. Alternatively, the first channel 130 can be used to allow fluid to flow along the first direction x in the following ways: the parts on the first channel 130 for adding fluid and the parts for discharging fluid are roughly distributed along the first direction x, thereby allowing fluid entering the first channel 130 to be considered as flowing along the first direction x.

[0051] Similarly, the second channel 140 can be used to allow fluid to flow along the first direction x in the following way: the second channel 140 extends along the first direction x, thereby allowing fluid within the second channel 140 to flow along the first direction x. Alternatively, the second channel 140 can be used to allow fluid to flow along the first direction x in the following way: the parts on the second channel 140 for adding fluid and the parts for discharging fluid are roughly distributed along the first direction x, thereby allowing fluid entering the second channel 140 to be considered as flowing along the first direction x.

[0052] In this embodiment of the application, the first channel 130 extends along the first direction x, and the parts for adding fluid and the parts for discharging fluid on the second channel 140 are distributed along the first direction x.

[0053] Combination Figure 2 and Figure 3 The partition 120 includes a connecting section 123, which includes a first protrusion 1231 and a second protrusion 1232 extending along a first direction x. The first protrusion 1231 and the second protrusion 1232 are spaced apart along a second direction y. The sidewalls of the first protrusion 1231 and the second protrusion 1232 that are opposite to each other along the second direction y are connected to the main body 110. A connecting port 150 is formed between the first protrusion 1231 and the second protrusion 1232, which connects the first channel 130 and the second channel 140. The second direction y is set at an angle to the first direction x.

[0054] Wherein, the sidewalls of the first protrusion 1231 and the second protrusion 1232 that are opposite to each other along the second direction y are connected to the main body 110 as follows: the sidewall of the first protrusion 1231 that is opposite to the sidewall of the second protrusion 1232 along the second direction y is connected to the main body 110, and the sidewall of the second protrusion 1232 that is opposite to the sidewall of the first protrusion 1231 along the second direction y is connected to the main body 110.

[0055] Combination Figure 4The first protrusion 1231 and the second protrusion 1232 of the partition 120 form an edge structure. The presence of the edge structure changes the spreading direction of the fluid in the first channel 130, restricting the spread of the fluid on the solid surface and preventing the fluid in the first channel 130 from diffusing and overflowing into the second channel 140 through the connecting port 150. The fluid in the first channel 130 is confined within the first channel 130, and a gas-liquid interface is formed at the connecting port 150 to constitute an organ-blood vessel interface between the first channel 130 and the second channel 140, facilitating the exchange of nutrients, gases, and metabolites. This allows for the cultivation of vascular endothelial cells and corresponding organ cells in the first channel 130 and the second channel 140 respectively, without the need for a separate porous membrane, resulting in a simple structure and convenient operation.

[0056] It should be noted that the scheme of forming an edge structure by the first protrusion 1231 and the second protrusion 1232 of the partition 120 and using the gas-liquid interface formed at the edge structure to realize the exchange of nutrients, gases and metabolites in the fluids of the two channels in the embodiment of this application can be directly applied to the existing organ-on-a-chip. It is only necessary to set the first protrusion 1231 and the second protrusion 1232 proposed in this embodiment between the two channels of the existing organ-on-a-chip, while other structures of the existing organ-on-a-chip do not need to be adjusted. It has strong adaptability and helps to reduce research and development costs.

[0057] Optionally, the second direction y is set at an angle to the first direction x. The angle between the second direction y and the first direction x can be any angle, such as 0°, 45°, 90°, 135°, etc. Preferably, the angle between the second direction y and the first direction x can be 90°. Preferably, the second direction y is parallel to the first sidewall 121 of the partition member 120.

[0058] Optionally, the cross-section of the first channel 130 may be approximately a first rectangle, and the first sidewall 121 may correspond to one side of the first rectangle. In addition to the first sidewall 121, the inner wall surface of the first channel 130 also includes a third sidewall 131 and a fourth sidewall 132 extending along the first direction x and located on opposite sides of the first sidewall 121, respectively. The third sidewall 131 may be parallel to the fourth sidewall 132 and both may be perpendicular to the first sidewall 121.

[0059] Optionally, the cross-section of the second channel 140 can be approximately a second rectangle, and the second sidewall 122 can correspond to one side of the second rectangle. In addition to the second sidewall 122, the inner wall of the second channel 140 also includes a fifth sidewall 141 and a sixth sidewall 142 extending along the first direction x and located on opposite sides of the second sidewall 122. The fifth sidewall 141 can be parallel to the sixth sidewall 142, and both are perpendicular to the second sidewall 122. Optionally, the second sidewall 122 can be parallel to the first sidewall 121. The fifth sidewall 141 can be in the same plane as the third sidewall 131, and the sixth sidewall 142 can be in the same plane as the fourth sidewall 132.

[0060] Optionally, the width of the first channel 130 along the third direction z is greater than or equal to 200 μm and less than or equal to 600 μm, the width of the second channel 140 along the third direction z is greater than or equal to 300 μm and less than or equal to 700 μm, and the height of the first channel 130 and the second channel 140 along the second direction y is greater than or equal to 200 μm and less than or equal to 400 μm. The first direction x, the second direction y, and the third direction z are all perpendicular to each other.

[0061] Optionally, the cross-section of the first protrusion 1231 can be approximately rectangular, and the cross-section of the second protrusion 1232 can be approximately rectangular. The height of the first protrusion 1231 and the second protrusion 1232 along the second direction y is greater than or equal to 50 μm and less than or equal to 200 μm, and the width of the first protrusion 1231 and the second protrusion 1232 along the third direction z is greater than or equal to 100 μm and less than or equal to 200 μm. The height of the first protrusion 1231 along the second direction y can be approximately equal to the height of the second protrusion 1231 along the second direction y, and the width of the first protrusion 1231 along the third direction z can be approximately equal to the width of the second protrusion 1232 along the third direction z. This embodiment of the application does not limit this.

[0062] Combination Figure 4 and Figure 5 The first protrusion 1231 has a first sub-sidewall 1233 and a second sub-sidewall 1234 extending along the first direction x and facing away from each other, and the second protrusion 1232 has a third sub-sidewall 1235 and a fourth sub-sidewall 1236 extending along the first direction x and facing away from each other. The first sidewall 121 includes the first sub-sidewall 1233 and the third sub-sidewall 1235, and the second sidewall 122 includes the second sub-sidewall 1234 and the fourth sub-sidewall 1236.

[0063] Optionally, the second sub-sidewall 1234 and the fourth sub-sidewall 1236 are located on the same plane, so that the gas-liquid interface at the communication port 150 can be formed under the premise that the surface of the second sidewall 122 is smooth, which is conducive to the fluid flow in the second channel 140.

[0064] Optionally, the first sub-sidewall 1233 and the third sub-sidewall 1235 are located in the same plane to make the surface of the first sidewall 121 smooth, which facilitates fluid flow in the first channel 130. Optionally, the second sub-sidewall 1234 and the fourth sub-sidewall 1236 are both perpendicular to the fifth sidewall 141, and the first sub-sidewall 1233 and the third sub-sidewall 1235 are both perpendicular to the fifth sidewall 141.

[0065] Optionally, combined Figure 5 The first protrusion 1231 may be a ridge extending along the first direction x, and the first protrusion 1231 may also include a plurality of first sub-protrusions spaced apart along the first direction x. Optionally, the second protrusion 1232 may be a ridge extending along the first direction x, and the second protrusion 1232 may also include a plurality of second sub-protrusions spaced apart along the first direction x.

[0066] Optionally, the partition 120 further includes a blocking section 124, which is distributed along the first direction x with the connecting section 123. The blocking section 124 disconnects the connection between the first channel 130 and the second channel 140. When the fluid in the first channel 130 and the fluid in the second channel 140 flow at the position corresponding to the blocking section 124, they are disconnected by the blocking section 124. However, when the fluid flows at the position corresponding to the connecting section 123, they can exchange nutrients, gases, metabolites, etc., through the connecting port 150 at the connecting section 123.

[0067] Combination Figure 5 The partition 120 may include two blocking sections 124, which are distributed along a first direction x on opposite sides of the connecting section 123. Optionally, the blocking section 124 has a fifth sub-sidewall 1241 and a sixth sub-sidewall 1242 extending along the first direction x and facing away from each other, the first sidewall 121 including the fifth sub-sidewall 1241 and the second sidewall 122 including the sixth sub-sidewall 1242.

[0068] In one exemplary embodiment, in the direction from one end of the blocking segment 124 to the other end of the connecting segment 123, the distance between the fifth sub-sidewall 1241 and the sixth sub-sidewall 1242 gradually increases. That is, in the direction from one end of the blocking segment 124 to the other end of the connecting segment 123, the distance between the channel segment of the first channel 130 corresponding to the blocking segment 124 and the channel segment of the second channel 140 corresponding to the blocking segment 124 gradually increases. This reduces interference between the two channels and facilitates the processing design of the two channels.

[0069] Optionally, the included angle between the fifth sub-sidewall 1241 and the sixth sub-sidewall 1242 can be greater than or equal to 5° and less than or equal to 67.5°. For example, the included angle between the fifth sub-sidewall 1241 and the sixth sub-sidewall 1242 can be 5°, 7°, 45°, 67.5°, etc., and this application embodiment does not limit this.

[0070] Optionally, the first sub-sidewall 1233 of the first protrusion 1231 can be smoothly connected to the fifth sub-sidewall 1241 of the blocking section 124, and the second sub-sidewall 1234 of the first protrusion 1231 can be smoothly connected to the sixth sub-sidewall 1242 of the blocking section 124, so as to improve the smoothness of the connection between the fluid in the first channel 130 and the fluid in the second channel 140 at the junction of the blocking section 124 and the connecting section 123.

[0071] Optionally, the third sub-sidewall 1235 of the second protrusion 1232 can be smoothly connected to the fifth sub-sidewall 1241 of the blocking section 124, and the fourth sub-sidewall 1236 of the second protrusion 1232 can be smoothly connected to the sixth sub-sidewall 1242 of the blocking section 124, so as to improve the smoothness of the connection between the fluid in the first channel 130 and the fluid in the second channel 140 at the junction of the blocking section 124 and the connecting section 123.

[0072] Optionally, combined Figure 4 and Figure 5 The cell culture chip 100 includes two partitions 120. The first sidewalls 121 of the two partitions 120 are arranged opposite each other, forming a first channel 130 between the two first sidewalls 121. The second sidewalls 122 of the two partitions 120 are arranged opposite each other, and a second channel 140 is formed on one side of the second sidewall 122 of each partition 120. That is, a second channel 140 is provided on each side of the first channel 130, so that nutrients, gases, and metabolites can be exchanged on both sides of the material in the first channel 130 through the two second channels 140.

[0073] Optionally, please combine Figure 2 The main body 110 includes a first plate 111 and a second plate 112, which are stacked on top of each other. A partition 120 is located between the first plate 111 and the second plate 112. A first protrusion 1231 connects to the first plate 111, and a second protrusion 1232 connects to the second plate 112. The second plate 112 also has a first through hole 1121 communicating with the first channel 130, a second through hole 1122 communicating with the first channel 130, a third through hole 1123 communicating with the second channel 140, and a fourth through hole 1124 communicating with the second channel 140. The first through hole 1121 and the second through hole 1122 can be located at opposite ends of the first channel 130, and the third through hole 1123 and the fourth through hole 1124 can be located at opposite ends of the second channel 140.

[0074] Optionally, the first channel 130 may have an enlarged portion corresponding to the first through hole 1121 and / or the second through hole 1122 to facilitate communication with the first through hole 1121 and / or the second through hole 1122. Optionally, the second channel 140 may have an enlarged portion corresponding to the third through hole 1123 and / or the fourth through hole 1124 to facilitate communication with the third through hole 1123 and / or the fourth through hole 1124.

[0075] Optionally, the first plate 111 and / or the second plate 112 may be hydrophilic surface treated to improve the fluid flow within the first channel 130 and the second channel 140.

[0076] Optionally, the main body 110 may further include a third plate 113, which is stacked on the side of the second plate 112 away from the first plate 111. The third plate 113 has a first inlet 1131 communicating with the first through hole 1121, a first outlet 1132 communicating with the second through hole 1122, a second inlet 1133 communicating with the third through hole 1123, and a second outlet 1134 communicating with the fourth through hole 1124.

[0077] Optionally, the third plate 113 is provided with at least one of the following: a first liquid storage tank corresponding to the first inlet 1131, a second liquid storage tank corresponding to the second inlet 1133, a third liquid storage tank corresponding to the first outlet 1132, and a fourth liquid storage tank corresponding to the second outlet 1134. Optionally, the cross-section of each liquid storage tank is approximately fan-shaped.

[0078] Optionally, the cell culture chip 100 can be made of materials such as PMMA, PC, COC, COP, and PS. Optionally, the first protrusion 1231 and the blocking segment can be integrally formed with the first plate 111, and the second protrusion 1232 can be integrally formed with the second plate 112, in order to reduce the manufacturing complexity and cost of the cell culture chip 100, enhance the fluid confinement effect in the chip, and improve the success rate of constructing the membrane-free cell culture chip 100. The integral forming can be achieved through injection molding only; this embodiment does not limit this. Optionally, the first plate 111 and the second plate 112, and the second plate 112 and the third plate 113, can be bonded and assembled by means of hot pressing, ultrasound, laser, adhesive, etc.

[0079] Optionally, in the embodiments of this application, the flow of fluid in the first channel 130 and / or the flow of fluid in the second channel 140 can be driven by a pumpless gravity drive, an external peristaltic pump drive, an injection pump drive, etc., so that the fluid can provide the fluid shear force environment required for cell growth.

[0080] Through simulation software verification, when the partition 120 of the cell culture chip 100 includes a first protrusion 1231 and a second protrusion 1232 distributed at intervals along the second direction y, compared to setting the partition 120 to include a single protrusion structure distributed at intervals along the second direction y, the maximum pressure that the gas-liquid interface formed between the first channel 130 and the second channel 140 can withstand can be greatly increased, and the fluid in the first channel 130 can be better restricted.

[0081] Specifically, in the case where the cell culture chip 100 includes one first channel 130 and two second channels 140, if the width of the first channel 130 and the second channel 140 along the third direction z is 400 μm, the height of the first channel 130 and the second channel 140 along the second direction y is 400 μm, the contact angle of the first plate 111 is 100°, the contact angle of the second plate 112 is 60°, the width of the first protrusion 1231 and the second protrusion 1232 along the third direction z is 100 μm, and the height of the first protrusion 1231 and the second protrusion 1232 along the second direction y is 100 μm, then, combined with Figures 6 to 9 Through simulation software verification, when the partition 120 is configured to include a protrusion structure spaced apart along the second direction y, the maximum pressure that the gas-liquid interface formed between the first channel 130 and the second channel 140 can withstand is 20 Pa. However, when the partition 120 in this embodiment includes a first protrusion 1231 and a second protrusion 1232 spaced apart along the second direction y, the maximum pressure that the gas-liquid interface formed between the first channel 130 and the second channel 140 can withstand is 200 Pa. That is, the blocking effect of the double protrusion in this embodiment is better.

[0082] The simulation software can be modeled using COMSOL Multiphysics (5.6). During the simulation verification, it can be assumed that water and air are incompressible Newtonian fluids and that their density is constant.

[0083] Specifically, the maximum pressure that the gas-liquid interface can withstand can satisfy the Navier-Stokes equation, which can be expressed as:

[0084]

[0085]

[0086] In the formula: U is the flow velocity, ρ is the density, μ is the dynamic viscosity, p is the pressure, and F is the flow rate. st Let I represent surface tension and I be the identity matrix. During modeling, the initial conditions, flow velocity U and pressure p, are both set to 0. The left side represents the pressure boundary (0), and the right side represents the pressure inlet boundary. By changing these values, the maximum pressure that different structures can withstand can be determined, thus assessing their performance.

[0087] The numerical simulation method is as follows: the phase-field method is used to trace the gas-liquid interface. The entire computational domain is divided into an air region Ω1 (-1≤φ<0) and a water region Ω2 (0<φ≤1) by a higher-order function φ. The region where φ=0 is the gas-liquid interface.

[0088] Among them, the variable φ satisfies the Cahn-Hilliard equation, which can be expressed as:

[0089]

[0090] In the formula: γ represents mobility, which determines the timescale of Cahn-Hilliard diffusion; λ represents the mixing energy density; and ε represents the interface thickness. The above equations, boundary conditions, and initial values ​​are all implemented in the COMSOL Multiphysics (5.6) finite element framework. Furthermore, the relative tolerance (A... rt =0.005) and absolute tolerance (A at,k =0.05) controls the error in the calculation step. The next calculation will only proceed when the error meets the following conditions:

[0091]

[0092] In the formula: M is the number of physical fields, N j V is the number of degrees of freedom in a physical field. k E is the solution vector for a specific time step. k This represents the error in the calculator.

[0093] The cell culture chip 100 of this embodiment is used in the following way for intestinal microarray applications:

[0094] S11. The second plate 112 of the main body 110 is hydrophilically treated and assembled with the first plate 111 and the third plate 113 to form a cell culture chip 100.

[0095] S12. The cell culture chip 100 is sterilized.

[0096] S13. Matrigel / collagen mixed hydrogel is added into the first channel 130. Due to the blocking effect of the first protrusion 1231 and the second protrusion 1232, the Matrigel / collagen mixed hydrogel is confined to the second channel 140 and will not flow into the first channel 130. The Matrigel / collagen mixed hydrogel can be added to the first channel 130 through the first inlet 1131, and excess Matrigel / collagen mixed hydrogel can flow out through the first outlet 1132.

[0097] S14, Matrigel / collagen mixed hydrogel solidifies to form hydrogel strips.

[0098] S15. Intestinal epithelial cells are added to one of the second channels 140, allowing them to adhere to the Matrigel / collagen mixed hydrogel at the gas-liquid interface between the first channel 130 and the second channel 140. Intestinal epithelial cell culture medium is added to one of the second channels 140 to provide the necessary nutrients for cell growth. The intestinal epithelial cells and culture medium can be added through the second inlet 1133 corresponding to the second channel 140, and excess cells and culture medium can flow out through the second outlet 1134 corresponding to the second channel 140.

[0099] S16. Add vascular endothelial cells to another second channel 140, allowing the vascular endothelial cells to adhere to the Matrigel / collagen mixed hydrogel gas-liquid interface between the first channel 130 and the second channel 140. Add vascular endothelial cell culture medium to this second channel 140 to provide the nutrients required for the growth of the vascular endothelial cells. The vascular endothelial cells and vascular endothelial cell culture medium can be added through the second inlet 1133 corresponding to this second channel 140, and excess vascular endothelial cells and vascular endothelial cell culture medium can flow out through the second outlet 1134 corresponding to this second channel 140.

[0100] S17. After successful cell culture, the intestinal-endothelial organ model is constructed for subsequent testing.

[0101] The cell culture chip 100 of this embodiment is used in the following way for 3D cell culture:

[0102] S21. The second plate 112 of the main body 110 is hydrophilically treated and assembled with the first plate 111 and the third plate 113 to form a cell culture chip 100.

[0103] S22. The cell culture chip 100 is sterilized.

[0104] S23. Add a hydrogel mixed with cells into the first channel 130. Due to the blocking effect of the first protrusion 1231 and the second protrusion 1232, the hydrogel mixed with cells is confined to the second channel 140 and will not flow into the first channel 130. The hydrogel mixed with cells can be added into the first channel 130 through the first inlet 1131, and excess hydrogel mixed with cells can flow out through the first outlet 1132.

[0105] S24. The hydrogel containing cells solidifies to form hydrogel strips.

[0106] S25. Add cell culture medium to the two second channels 140 to provide the nutrients required for cell growth. The cell culture medium can be added from the second inlet 1133 corresponding to the second channel 140, and excess cell culture medium can flow out from the second outlet 1134 corresponding to the second channel 140.

[0107] S26. Cell culture was successful and is ready for subsequent testing.

[0108] Example 2

[0109] Please see Figures 10 to 13 This embodiment provides another cell culture chip 100, which is substantially the same as the cell culture chip 100 in Embodiment 1. The difference is that in Embodiment 1, the second sub-sidewall 1234 of the first protrusion 1231 and the fourth sub-sidewall 1236 of the second protrusion 1232 are located on the same plane, while in this embodiment, combined with Figure 10 and Figure 11 At least a portion of the fourth sub-sidewall 1236 of the second protrusion 1232 is located on the side of the second sub-sidewall 1234 of the first protrusion 1231 away from the first sub-sidewall 1233 of the first protrusion 1231.

[0110] Experiments show that placing at least a portion of the fourth sub-sidewall 1236 on the side of the second sub-sidewall 1234 away from the first sub-sidewall 1233 can control the morphology of the gas-liquid interface formed at the communication port 150 within the first channel 130 and the resulting surface energy. Furthermore, as the distance between the second sub-sidewall 1234 and the fourth sub-sidewall 1236 along the third z-direction increases, the gas-liquid interface becomes larger, and the Gibbs free energy of the fluid surface increases, which can further counteract the pressure generated inside the fluid within the first channel 130, achieving a better fluid confinement effect.

[0111] Optionally, the distance between the second sub-sidewall 1234 and the fourth sub-sidewall 1236 along the third direction z is greater than or equal to 50 μm and less than or equal to 400 μm. Optionally, combined with Figure 12 At least part of the third sub-sidewall 1235 is located on the side of the second sub-sidewall 1234 away from the first sub-sidewall 1233, which increases the distance between the first protrusion 1231 and the second protrusion 1232 along the third direction z, thereby improving the fluid restriction effect in the first channel 130.

[0112] Combination Figure 10 The second protrusion 1232 can extend in a straight line, which makes the structure of the second protrusion 1232 simple and easier to manufacture.

[0113] Combination Figure 13The second protrusion 1232 may also include a middle portion 1237 distributed along the first direction x and an end portion 1238 connecting the middle portion 1237. The middle portion 1237 is located on the side of the second sub-sidewall 1234 away from the first sub-sidewall 1233, and the end portion 1238 bends and extends toward the first sub-sidewall 1233, making the change of the second protrusion 1232 more gradual, which is conducive to the smooth flow of fluid in the first channel 130 and the second channel 140.

[0114] Optionally, combined Figure 10 The third sub-sidewall 1235 of the second protrusion 1232 may be located between the fifth sub-sidewall 1241 and the sixth sub-sidewall 1242 of the blocking section 124, and the fourth sub-sidewall 1236 of the second protrusion 1232 may be located on the side of the sixth sub-sidewall 1242 of the blocking section 124 away from the fifth sub-sidewall 1241. Optionally, the third sub-sidewall 1235 of the second protrusion 1232 may be located on the side of the sixth sub-sidewall 1242 of the blocking section 124 away from the fifth sub-sidewall 1241, and the fourth sub-sidewall 1236 of the second protrusion 1232 may also be located on the side of the sixth sub-sidewall 1242 of the blocking section 124 away from the fifth sub-sidewall 1241.

[0115] Through simulation software verification, when the partition 120 of the cell culture chip 100 includes a first protrusion 1231 and a second protrusion 1232 distributed at intervals along the second direction y, placing at least a portion of the fourth sub-sidewall 1236 of the second protrusion 1232 on the side of the second sub-sidewall 1234 of the first protrusion 1231 away from the first sub-sidewall 1233 of the first protrusion 1231, compared with the first embodiment where the second sub-sidewall 1234 of the first protrusion 1231 and the fourth sub-sidewall 1236 of the second protrusion 1232 are located on the same plane, the maximum pressure that can be withstood at the gas-liquid interface formed between the first channel 130 and the second channel 140 can be greatly increased, and the fluid in the first channel 130 can be better restricted.

[0116] Specifically, in the case where the cell culture chip 100 includes one first channel 130 and two second channels 140, if the width of the first channel 130 and the second channel 140 along the third direction z is 400 μm, the height of the first channel 130 and the second channel 140 along the second direction y is 400 μm, the contact angle of the first plate 111 is 100°, the contact angle of the second plate 112 is 60°, the width of the first protrusion 1231 and the second protrusion 1232 along the third direction z is 100 μm, and the height of the first protrusion 1231 and the second protrusion 1232 along the second direction y is 100 μm, then the structure... Figure 14 and Figure 15Through simulation software verification, when at least a portion of the fourth sub-sidewall 1236 of the second protrusion 1232 is located on the side of the second sub-sidewall 1234 of the first protrusion 1231 away from the first sub-sidewall 1233 of the first protrusion 1231, the maximum pressure that the gas-liquid interface formed between the first channel 130 and the second channel 140 can withstand is 220 Pa, and the blocking effect is better.

[0117] The cell culture chip 100 of this embodiment is used in the following way for intestinal microarray applications:

[0118] S31. The second plate 112 of the main body 110 is hydrophilically treated and assembled with the first plate 111 and the third plate 113 to form a cell culture chip 100.

[0119] S32. The cell culture chip 100 is sterilized.

[0120] S33. Matrigel / collagen mixed hydrogel is added into the first channel 130. Due to the blocking effect of the first protrusion 1231 and the second protrusion 1232, the Matrigel / collagen mixed hydrogel is confined to the second channel 140 and will not flow into the first channel 130. The Matrigel / collagen mixed hydrogel can be added to the first channel 130 through the first inlet 1131, and excess Matrigel / collagen mixed hydrogel can flow out through the first outlet 1132.

[0121] S34, Matrigel / collagen mixed hydrogel solidifies to form hydrogel strips.

[0122] S35. Intestinal epithelial cells are added to one of the second channels 140, allowing them to adhere to the Matrigel / collagen mixed hydrogel at the gas-liquid interface between the first channel 130 and the second channel 140. Intestinal epithelial cell culture medium is added to one of the second channels 140 to provide the necessary nutrients for cell growth. The intestinal epithelial cells and culture medium can be added through the second inlet 1133 corresponding to the second channel 140, and excess cells and culture medium can flow out through the second outlet 1134 corresponding to the second channel 140.

[0123] S36. Add vascular endothelial cells to another second channel 140, allowing the vascular endothelial cells to adhere to the Matrigel / collagen mixed hydrogel gas-liquid interface between the first channel 130 and the second channel 140. Add vascular endothelial cell culture medium to this other second channel 140 to provide the nutrients required for the growth of the vascular endothelial cells. The vascular endothelial cells and vascular endothelial cell culture medium can be added through the second inlet 1133 corresponding to this other second channel 140, and excess vascular endothelial cells and vascular endothelial cell culture medium can flow out through the second outlet 1134 corresponding to this other second channel 140.

[0124] S37. After successful cell culture, the intestinal-endothelial organ model was constructed for subsequent testing.

[0125] The cell culture chip 100 of this embodiment is used in the following way for 3D cell culture:

[0126] S41. The second plate 112 of the main body 110 is hydrophilically treated and assembled with the first plate 111 and the third plate 113 to form a cell culture chip 100.

[0127] S42. The cell culture chip 100 is sterilized.

[0128] S43. Add a cell-mixed hydrogel into the first channel 130. Due to the blocking effect of the first protrusion 1231 and the second protrusion 1232, the cell-mixed hydrogel is confined to the second channel 140 and will not flow into the first channel 130. The cell-mixed hydrogel can be added into the first channel 130 through the first inlet 1131, and excess cell-mixed hydrogel can flow out through the first outlet 1132.

[0129] S44. The hydrogel containing cells solidifies to form hydrogel strips.

[0130] S45. Add cell culture medium to the two second channels 140 to provide the nutrients required for cell growth. The cell culture medium can be added from the second inlet 1133 corresponding to the second channel 140, and excess cell culture medium can flow out from the second outlet 1134 corresponding to the second channel 140.

[0131] S46. Cell culture was successful and is ready for subsequent testing.

[0132] In the description of this application, it should be understood that the terms "first," "second," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances. Furthermore, in the description of this application, unless otherwise stated, "multiple" refers to two or more. "And / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, or B existing alone. The character " / " generally indicates that the preceding and following related objects are in an "or" relationship.

[0133] The above-disclosed embodiments are merely preferred embodiments of this application and should not be construed as limiting the scope of this application. Therefore, any equivalent variations made in accordance with the claims of this application shall still fall within the scope of this application.

Claims

1. A cell culture chip, characterized in that, include: The main body includes a first plate and a second plate, wherein the second plate and the first plate are stacked together. A partition member is located within the main body between and connected to the main body, between the first plate and the second plate. The partition member has opposing first and second sidewalls. A first channel is formed between the partition member and the main body on the side of the first sidewall, and a second channel is formed on the side of the second sidewall. The first channel is used for fluid flow in a first direction, and the second channel is used for fluid flow in the first direction. The partition member includes a connecting section and two blocking sections. The connecting section includes a first protrusion and a second protrusion extending along the first direction. The first protrusion and the second protrusion are spaced apart along a second direction. The protrusions, with their opposite sidewalls along the second direction, connect the first plate and the second plate respectively. A connecting port is formed between the first protrusion and the second protrusion, connecting the first channel and the second channel. The second direction is set at an angle to the first direction. The first protrusion has a rectangular cross-section and is a convex strip extending along the first direction. The second protrusion has a rectangular cross-section and is a convex strip extending along the first direction. Two blocking segments are distributed on opposite sides of the connecting segment along the first direction. The blocking segments separate the first channel and the second channel so that the first channel and the second channel are connected only through the connecting port. The inner wall of the first channel includes a third sidewall and a fourth sidewall extending along the first direction and located on opposite sides of the first sidewall; the inner wall of the second channel includes a fifth sidewall and a sixth sidewall extending along the first direction and located on opposite sides of the second sidewall; the fifth sidewall and the third sidewall are located in the same plane, and the sixth sidewall and the fourth sidewall are located in the same plane. The first protrusion has a first sub-sidewall and a second sub-sidewall extending along the first direction and opposite to each other, and the second protrusion has a third sub-sidewall and a fourth sub-sidewall extending along the first direction and opposite to each other. The first sidewall includes the first sub-sidewall and the third sub-sidewall, and the second sidewall includes the second sub-sidewall and the fourth sub-sidewall. At least a portion of the fourth sub-sidewall is located on the side of the second sub-sidewall away from the first sub-sidewall.

2. The cell culture chip according to claim 1, characterized in that, At least part of the third sub-sidewall is located on the side of the second sub-sidewall away from the first sub-sidewall.

3. The cell culture chip according to claim 1, characterized in that, The second protrusion includes a central portion distributed along the first direction and an end portion connecting the central portion, the central portion being located on the side of the second sub-sidewall facing away from the first sub-sidewall, and the end portion extending in a direction bent towards the first sub-sidewall; or The second protrusion extends in a straight line.

4. The cell culture chip according to claim 1, characterized in that, The barrier segment has a fifth sub-sidewall and a sixth sub-sidewall extending along the first direction and facing away from each other, the first sidewall including the fifth sub-sidewall, and the second sidewall including the sixth sub-sidewall. In the direction from one end of the blocking segment to the other end of the connecting segment, the distance between the fifth sub-sidewall and the sixth sub-sidewall gradually increases.

5. The cell culture chip according to claim 4, characterized in that, The angle between the fifth sub-sidewall and the sixth sub-sidewall is greater than or equal to 5° and less than or equal to 67.5°.

6. The cell culture chip according to claim 1, characterized in that, The partition member forms the first channel and the second channel with the first plate and the second plate. The second plate also has a first through hole communicating with the first channel, a second through hole communicating with the first channel, a third through hole communicating with the second channel and a fourth through hole communicating with the second channel.

7. The cell culture chip according to claim 6, characterized in that, The subject also includes: The third plate is stacked on the side of the second plate away from the first plate. The third plate has a first inlet communicating with the first through hole, a first outlet communicating with the second through hole, a second inlet communicating with the third through hole, and a second outlet communicating with the fourth through hole.

8. The cell culture chip according to any one of claims 1-7, characterized in that, The cell culture chip includes two partitions, the first sidewalls of the two partitions are arranged opposite each other and a first channel is formed between the two first sidewalls, the second sidewalls of the two partitions are arranged opposite each other, and a second channel is formed on one side of the second sidewall of each partition.

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