Cell culture chip and single-cycle fluidic device

Abstract

The application discloses a cell culture chip and a single-cycle fluid device, wherein the cell culture chip comprises a base, and the base is provided with a plurality of liquid storage chambers used for storing culture solution, each of the liquid storage chambers is provided with an outlet and an inlet, the outlet only allows the culture solution to flow out, and the inlet only allows the culture solution to flow in, at least one culture groove is arranged between each pair of the liquid storage chambers, and the culture groove is used for culturing a cell chip; in each pair of the liquid storage chambers, the outlet of one of the liquid storage chambers is communicated with the inlet of the other liquid storage chamber through the at least one culture groove, and the inlet of the one liquid storage chamber is communicated with the outlet of the other liquid storage chamber. By adopting the two single-flow liquid storage chambers, the culture solution in the two liquid storage chambers can only flow out from the outlet and flow in from the inlet, so that the unidirectional flow of the culture solution in the culture groove is realized, and the culture solution in the two liquid storage chambers can also be circulated.

Description

Technical Field

[0001] This invention relates to the field of cell or organ chip culture, and particularly to a cell culture chip and a single-circulation fluid device. Background Technology

[0002] In the field of biochip culture, such as cell or organ culture, fluids can be used to flow on the cell chip to simulate the environment of human cells or organs under blood flow. Currently, most traditional gravity-driven solutions involve placing cells or organ-on-a-chip on a shaker and using pendulum-like or seesaw-like shaking to make the culture medium flow back and forth on the cells or organ-on-a-chip, thus simulating the effect of blood flow. For example, the shaker and culture chamber disclosed in the patent application No. 202110791120.8, entitled "A Method for Constructing a Model of Adeno-Associated Virus Crossing the Blood-Brain Barrier," are examples of such solutions. However, this flow is a back-and-forth movement in two directions, which does not match the unidirectional circulation of blood and other bodily fluids in real organisms. In addition, there are a few culture devices that can achieve unidirectional flow, such as those using a drive pump to achieve single-cycle flow. For example, the patent application No. 202110823992.8, entitled "Microfluidic Cell Chip and Virus Isolation and Culture Method Based on the Cell Chip," discloses the use of a peristaltic pump to achieve unidirectional flow. However, this requires an additional drive pump, resulting in higher costs. Summary of the Invention

[0003] The main objective of this invention is to provide a cell culture chip and a single-circulation fluid device, aiming to solve the problems mentioned in the background art.

[0004] To achieve the above objectives, the present invention proposes a cell culture chip, comprising:

[0005] The base is provided with at least one pair of oppositely arranged liquid storage chambers for storing culture medium. Each liquid storage chamber is provided with an outlet and an inlet. The outlet allows only the culture medium to flow out, and the inlet allows only the culture medium to flow in. At least one culture tank is provided between each pair of liquid storage chambers for culturing cells.

[0006] In each pair of storage chambers, the outlet of one storage chamber is connected to the inlet of the other storage chamber through the at least one culture tank, and the inlet of one storage chamber is connected to the outlet of the other storage chamber.

[0007] In one embodiment, each of the liquid storage chambers is provided with a check valve at its inlet;

[0008] In each pair of storage chambers, the outlet of one storage chamber is connected to the check valve of the inlet of the other storage chamber through the at least one culture tank, and the check valve of the inlet of one storage chamber is connected to the outlet of the other storage chamber.

[0009] In one embodiment, the height of the inlet of each of the liquid storage chambers is higher than the height of the outlet;

[0010] In each pair of storage chambers, the outlet of one storage chamber is connected to the inlet of the other storage chamber through the at least one culture tank, and the inlet of one storage chamber is connected to the outlet of the other storage chamber.

[0011] In one embodiment, the culture tank is provided with an inlet channel and an outlet channel at both ends. The inlet channel is used to connect to the outlet of the storage chamber located upstream in the direction of culture medium flow, and the outlet channel is used to connect to the inlet of the storage chamber located downstream in the direction of culture medium flow.

[0012] In one embodiment, the liquid outlet channel extends upward at an angle from the outlet of the culture tank to the inlet of the liquid storage chamber located downstream of the flow direction of the culture medium.

[0013] In one embodiment, the inner bottom of the culture tank extends obliquely from the inlet of the culture tank toward the inlet of the liquid storage chamber located on the outlet side, so that the liquid outlet channel and the inner bottom of the liquid storage chamber are on the same plane.

[0014] In one embodiment, the liquid outlet channel extends horizontally from the outlet of the culture tank to the vertical projection position of the liquid inlet of the storage chamber located downstream of the flow direction of the culture medium, and then communicates upward with the liquid inlet.

[0015] In one embodiment, at least one flow-blocking plate is provided at the inner bottom of the liquid outlet channel. The flow-blocking plate is located in the horizontal extension section of the liquid outlet channel and is inclined from the inner bottom of the liquid outlet channel in the direction of culture medium flow.

[0016] In one embodiment, the at least one flow-blocking plate includes a plurality of flow-blocking plates, which are spaced apart along the direction from the liquid storage chamber to the culture tank at the inner bottom of the liquid outlet channel, and the area of ​​the plurality of flow-blocking plates gradually decreases in the direction of liquid flow.

[0017] In one embodiment, the inner bottom of the liquid storage chamber is provided with a flow guide surface, and the liquid inlet is located on the flow guide surface or on the inner sidewall adjacent to the flow guide surface. The flow guide surface extends obliquely from the side of the liquid inlet to the side of the liquid outlet.

[0018] In one embodiment, the liquid inlet is disposed on the guide surface, and the liquid inlet is located at the edge of the guide surface away from the culture tank.

[0019] In one embodiment, the guide surface extends upward in an inverted cone shape.

[0020] In one embodiment, in each pair of liquid storage chambers, the two liquid outlets are respectively located in the areas where the side walls of the two liquid storage chambers are opposite each other.

[0021] In one embodiment, the two liquid outlets are respectively located at the bottom of the regions opposite to the sidewalls of the two liquid storage chambers.

[0022] In one embodiment, in each pair of liquid storage chambers, the outlets of both liquid storage chambers are connected to the inlet of the other liquid storage chamber through the at least one culture tank.

[0023] In one embodiment, the bottom of the culture tank is provided with a plurality of grooves spaced apart along the line connecting the two liquid storage chambers. The grooves are used to place cultured cells, and the height of the grooves is not higher than the outlet height of the liquid inlet channel in the culture tank.

[0024] In one embodiment, the groove extends from the bottom of the culture tank to the bottom surface of the base.

[0025] In one embodiment, the bottom surface of the base is provided with a positioning groove, and all the grooves are located in the positioning groove. The positioning groove is used to position the light-transmitting film from the bottom of the base.

[0026] In one embodiment, the base, including the groove, is made of a pre-defined area of ​​light-transmitting material.

[0027] In one embodiment, the at least one culture tank includes a plurality of culture tanks arranged side by side between the liquid outlet and the liquid inlet of the two liquid storage chambers.

[0028] In one embodiment, a cover plate is also included for covering the two liquid storage chambers and the culture tank.

[0029] In one embodiment, the cover plate has an observation window in the area covering the culture tank.

[0030] In one embodiment, the cover plate is made of a transparent material in the area covering the culture tank.

[0031] In one embodiment, the cover plate has a buckle on the plate surface that covers the liquid storage chamber, and when the cover plate is closed, the two liquid storage chambers have hooks at the positions corresponding to the buckles.

[0032] In one embodiment, the base is further provided with a fixing hole for fixing the base to the rocking device.

[0033] In one embodiment, the base is further provided with flow direction markings.

[0034] The present invention also proposes a single-circulation fluid device, comprising the cell culture chip and shaker described in any of the above claims.

[0035] In one embodiment, the cell culture chip is fixed to the shaker via the base.

[0036] The technical solution of this invention employs two unidirectional flow storage chambers, with a culture tank positioned between them. The outlet of one storage chamber is connected to the inlet of the other, while the outlet and inlet of the other storage chamber are connected through the culture tank. Thus, when the plane containing the two storage chambers and the culture tank is oscillating, the culture solution in the two storage chambers can only flow out through the outlet and in through the inlet, thereby achieving unidirectional flow of the culture solution within the culture tank. Furthermore, during continuous oscillation, the culture solution in the two storage chambers can also circulate unidirectionally. Attached Figure Description

[0037] To more clearly illustrate the technical solutions in the embodiments of the present invention 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 the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0038] Figure 1 This is a schematic diagram of the structure of a cell culture chip according to an embodiment of the present invention.

[0039] Figure 2 This is a schematic diagram of another embodiment of the cell culture chip of the present invention;

[0040] Figure 3 for Figure 1 A schematic diagram of another embodiment of the cell culture chip of the present invention;

[0041] Figure 4 for Figure 3 A top view of a cell culture chip;

[0042] Figure 5 for Figure 3 Right view of the first reservoir of the cell culture chip;

[0043] Figure 6 A schematic diagram of another embodiment of the cell culture chip of the present invention;

[0044] Figure 7 This is a cross-sectional view of an embodiment of the reservoir in the cell culture chip of the present invention;

[0045] Figure 8This is a cross-sectional view of another embodiment of the reservoir in the cell culture chip of the present invention;

[0046] Figure 9 This is a schematic diagram of another embodiment of the cell culture chip of the present invention;

[0047] Figure 10 for Figure 3 The second culture tank is located on the left side of the liquid outlet channel;

[0048] Figure 11 This is a schematic diagram of another embodiment of the cell culture chip of the present invention;

[0049] Figure 12 This is a schematic diagram of another embodiment of the cell culture chip of the present invention.

[0050] Explanation of icon numbers:

[0051] 100-Base, 110-Fixing hole, 111-Positioning groove, 200-Liquid storage chamber, 210-First liquid storage chamber, 211-First liquid inlet, 212-First liquid outlet, 213-First guide surface, 214-First horizontal bottom surface, 310-Second liquid storage chamber, 311-Second liquid inlet, 312-Second liquid outlet, 313-Second guide surface, 314-Second horizontal bottom surface, 400-Cultivation tank, 410-First cultivation tank, 420-Second cultivation tank, 430-Groove, 440-Liquid inlet channel, 450-Liquid outlet channel, 460-Inner bottom, 500-Reflux channel, 600-Cover plate, 610-Observation window.

[0052] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0053] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0054] It should be noted that if the embodiments of the present invention involve directional indicators (such as up, down, left, right, front, back, etc.), the directional indicators are only used to explain the relative positional relationship and movement of the components in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicators will also change accordingly.

[0055] Furthermore, if the embodiments of this invention involve descriptions such as "first" or "second," these descriptions are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of those features. Additionally, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. If the combination of technical solutions is contradictory or impossible to implement, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by this invention.

[0056] This invention proposes a cell culture chip that can be used to culture or test cells such as adherent cells or vascular cells, as well as to culture or test organs. This application does not limit whether the chip is used to culture cells or organs, or to perform culture testing or cell culture.

[0057] In embodiments of the present invention, such as Figure 1 As shown, the cell culture chip includes a base 100, on which at least one pair of oppositely arranged liquid storage chambers 200 are provided. At least one culture tank 400 is provided between each pair of liquid storage chambers 200. Both liquid storage chambers 200 are unidirectional liquid storage chambers 200, that is, the culture medium in the liquid storage chamber 200 can only flow in from the inlet and out from the outlet. For ease of description, the left-hand storage chamber is referred to as the first storage chamber 210, and its outlet and inlet are referred to as the first outlet 212 and the first inlet 211, respectively. The right-hand storage chamber is referred to as the second storage chamber 310, and its outlet and inlet are referred to as the second outlet 312 and the second inlet 311, respectively. The first storage chamber 210 and the second storage chamber 310 are both mounted on the base 100, which is used to mount the first storage chamber 210 and the second storage chamber 310, as well as the culture tank 400, to swing with the base 100. The first storage chamber 210 and the second storage chamber 310 are used to hold the culture medium for cultured cells, and the culture tank 400 is used to hold cultured cells. Because the culture medium in the two reservoirs 200 can only flow out through the outlet and in through the inlet, unidirectional flow of the culture medium within the culture tank 400 is achieved. Furthermore, the culture medium in the two reservoirs 200 can circulate back and forth, realistically simulating the unidirectional circulation of body fluids within a living organism. It should be noted that, in situations such as... Figure 1 In one embodiment, only a pair of oppositely arranged liquid storage chambers 200 are provided on the base 100, and only one culture tank 400 is provided in each pair of oppositely arranged liquid storage chambers. In other embodiments, multiple pairs of oppositely arranged liquid storage chambers 200 may be provided on the base 100, and at least one culture tank 400 may be provided between each pair of liquid storage chambers 200.

[0058] It should be noted that in some embodiments, it is not necessary to follow the same procedure. Figure 1 The shapes and arrangements shown are used to set up the liquid storage chamber and the culture tank. For example, the culture tank can be set in an arc shape or a spiral shape. Or, the liquid storage chamber and the culture tank can be set at different heights, such as the liquid storage chamber on one side being located under the bottom plate (concave downwards) and the liquid storage chamber on the other side being located above the base (convex upwards), and so on.

[0059] The following is about Figure 1 The illustration only describes one pair of liquid storage chambers. If multiple pairs of oppositely arranged liquid storage chambers are required, the method for setting up this pair of liquid storage chambers and culture tank can be referred to. In this embodiment, for the two liquid storage chambers 200, the height of their inlets is higher than the height of their outlets. The outlet of one liquid storage chamber 200 is connected to the inlet of the other liquid storage chamber 200 through the at least one culture tank 400, and the inlet of one liquid storage chamber 200 is connected to the outlet of the other liquid storage chamber 200. Specifically, the two ends of the culture tank 400 are respectively connected to the first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310. The first inlet 211 of the first liquid storage chamber 210 is directly connected to the second outlet 312 of the second liquid storage chamber 310. Thus, when the first liquid storage chamber 210, the second liquid storage chamber 310, and the culture tank 400 sway with the base 100, when the first liquid storage chamber 210 is at a higher position and the second liquid storage chamber 310 is at a lower position, the culture solution in the first liquid storage chamber 210 flows from the first outlet 212 of the first liquid storage chamber 210 to the culture tank 400. The cultured cells flow into the second inlet 311 of the second reservoir 310. During this process, since the height of the first inlet 211 of the first reservoir 210 is higher than the height of the first outlet 212, by controlling the appropriate amount of culture medium, only the first outlet 212 of the first reservoir 210 can flow out of the culture medium to the right, while the first inlet 211 will not flow out of the culture medium to the right. When the second reservoir 310 swings to a higher position and the first reservoir 210 swings to a lower position, the culture medium begins to flow from the second outlet 312 of the second reservoir 310 to the first inlet 211 of the first reservoir 210, that is, through... Figure 1The reflux channel 500 shown flows to the first storage chamber 210. Similarly, because the second inlet 311 is higher than the second outlet 312, no culture medium flows from the second inlet 311 of the second storage chamber 310 to the culture tank 400 at this time. When the first storage chamber 210 swings back to a higher position, culture medium begins to flow into the culture tank 400 again through the first outlet 212 of the first storage chamber 210, and this process repeats. In this embodiment, for the culture tank 400 located in the middle, there is always only one direction of culture medium flowing from the first outlet 212 on the left to the second inlet 311 on the right. Therefore, a unidirectional flow environment simulating the body fluids in a biological organism is achieved. Furthermore, the culture medium can circulate repeatedly within the first and second storage chambers 210 and 311 through the reflux channel 500, making it more convenient to use.

[0060] It should be noted that in the above embodiment, the culture tank 400 is positioned between the first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310, achieving a unidirectional flow environment from left to right. In other embodiments, the culture tank 400 can also be positioned between the first inlet 211 of the first liquid storage chamber 210 and the second outlet 312 of the second liquid storage chamber 310, and the first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310 can be connected, thus achieving a unidirectional flow environment from right to left. This technical solution is applicable to... The culture tank 400 is disposed between the first liquid outlet 212 of the first liquid storage chamber 210 and the second liquid inlet 311 of the second liquid storage chamber 310. It is not limited to being disposed between the first liquid inlet 211 of the first liquid storage chamber 210 and the second liquid outlet 312 of the second liquid storage chamber 310. Moreover, in other embodiments, the culture tank 400 may be disposed between the first liquid outlet 212 of the first liquid storage chamber 210 and the second liquid inlet 311 of the second liquid storage chamber 310, and also between the first liquid inlet 211 of the first liquid storage chamber 210 and the second liquid outlet 312 of the second liquid storage chamber 310. See the following embodiments for details.

[0061] In addition, as mentioned above Figure 1 In the illustrated embodiment, a culture tank 400 is provided only between the first liquid outlet 212 of the first liquid storage chamber 210 and the second liquid inlet 311 of the second liquid storage chamber 310. In other embodiments, multiple culture tanks 400 may be provided, such as... Figure 2As shown, in this embodiment, three culture tanks 400 are arranged between the first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310. The three culture tanks 400 are arranged side-by-side, and their inlets and outlets are connected to the first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310 via a three-way pipe. Therefore, during left-right swaying, all three culture tanks 400 can achieve a unidirectional flow environment from left to right. It should be noted that this technical solution does not limit the number of culture tanks 400, and multiple culture tanks 400 can be used as described above. Figure 2 The first liquid outlet 212 of the first liquid storage chamber 210 and the second liquid inlet 311 of the second liquid storage chamber 310 are arranged side by side as shown. Alternatively, they can be arranged side by side between the first liquid inlet 211 of the first liquid storage chamber 210 and the second liquid outlet 312 of the second liquid storage chamber 310.

[0062] In another embodiment, check valves can be installed at the inlets of the two storage chambers 200. The check valves are installed in a direction that allows the culture medium to enter the storage chamber 200 only through the inlet and not to flow out. Thus, the culture medium in the two storage chambers 200 can only flow from the outlet to the inlet of the other storage chamber 200, achieving unidirectional flow environment simulation. For example... Figure 1 As shown, check valves (not shown in the figure) are installed at both the first inlet 211 and the second inlet 311. After the first outlet 212 is connected to the culture tank 400, it is connected to the second inlet 311 through the check valve. The second outlet 312 is connected to the first inlet 211 through the check valve at the first inlet 211. Therefore, when the culture medium flows from the culture tank 400 into the second storage chamber 310, it will not flow back into the culture tank 400 because of the check valve at the second inlet 311. Similarly, if the culture medium flows from the second storage chamber 310 into the first storage chamber 210, it can only flow into the culture tank 400 through the first outlet 212. In this embodiment, there is no restriction on the height of the outlet and the inlet.

[0063] like Figure 3 , Figure 4As shown in Figure 3, in another embodiment, the outlets of the first liquid storage chamber 210 and the second liquid storage chamber 310 are both connected to the inlet of the other liquid storage chamber 200 through at least one culture tank 400. The first outlet 212 of the first liquid storage chamber 210 and the second inlet 311 of the second liquid storage chamber 310 are connected through the first culture tank 410, and the first inlet 211 of the first liquid storage chamber 210 and the second outlet 312 of the second liquid storage chamber 310 are connected through the second culture tank 420. Both the first culture tank 410 and the second culture tank 420 are located on the base 100. Therefore, when the first liquid storage chamber 210 is at a higher position and the second liquid storage chamber 310 is at a lower position, the first liquid storage chamber... The culture medium in the liquid chamber 210 flows from the first outlet 212 through the first culture tank 410 and then enters the second storage chamber 310 through the second inlet 311. At this time, there is no culture medium flowing in the second culture tank 420. When the second storage chamber 310 is at a higher position and the first storage chamber 210 is at a lower position, the culture medium in the second storage chamber 310 flows through the second outlet 312 through the second culture tank 420 and enters the first storage chamber 210 through the first inlet 211. The above process is repeated continuously with the swaying, so that the first culture tank 410 always simulates a unidirectional flow environment from left to right, and the second culture tank 420 always simulates a unidirectional flow environment from right to left. Similarly, in other embodiments, a plurality of first culture tanks 410 may be provided between the first liquid outlet 212 of the first liquid storage chamber 210 and the second liquid inlet 311 of the second liquid storage chamber 310, and a plurality of second culture tanks 420 may be provided between the first liquid inlet 211 of the first liquid storage chamber 210 and the second liquid outlet 312 of the second liquid storage chamber 310. The communication structure of the plurality of first culture tanks 410 and the plurality of second culture tanks 420 may refer to the following. Figure 2 Configure the structure shown.

[0064] like Figure 1 As shown, in another embodiment, the culture tank 400 is provided with an inlet channel 440 and an outlet channel 450 at both ends. The inlet channel 440 is used to connect to the outlet of the storage chamber located upstream in the flow direction of the culture medium, and the outlet channel 450 is used to connect to the inlet of the storage chamber located downstream in the flow direction of the culture medium. For example... Figure 1 In the embodiment shown, the liquid inlet channel 440 of the culture tank 400 is connected to the first liquid outlet 212 of the first liquid storage chamber 210 on the left, and the liquid outlet channel 450 of the culture tank 400 is connected to the second liquid inlet 311 of the second liquid storage chamber 310 on the right.

[0065] like Figure 11As shown, in another embodiment, the outlet channel 450 extends upward at an angle from the outlet of the culture tank 400 to the inlet of the storage chamber 200 located downstream of the culture medium flow direction. As shown in the figure, in this embodiment, the second inlet 311 of the second storage chamber 310 on the right side is located on the side wall. The outlet channel 450 of the culture tank 400 extends directly upward from the outlet of the culture tank 400 to the second inlet 311 located on the side wall. Therefore, during oscillation, when the upwardly angled outlet channel 450 oscillates towards the second storage chamber 310, all the culture medium in the outlet channel 450 can enter the second storage chamber 310, thus ensuring that no culture medium flows back from the outlet channel 450 to the culture tank 400. Of course, in other embodiments, the outlet channel 450 can also extend upward directly from the outlet of the culture tank, passing through the side wall of the second storage chamber 310, and then extending into the interior of the second storage chamber 310.

[0066] like Figure 12 As shown, in another embodiment, the inner bottom 460 of the culture tank 400 extends obliquely from the inlet of the culture tank 400 toward the inlet of the liquid storage chamber 200 located on the outlet side, so that the liquid outlet channel 450 and the inner bottom of the culture tank 400 are located on the same plane. In the previous embodiment, the outlet channel 450 extends upward from the outlet of the culture tank 400 to the inlet of the storage chamber 200 downstream of the culture medium flow direction. In this embodiment, the inner bottom 460 of the culture tank 400 is set as an inclined surface and tilts directly from the inlet of the culture tank 400 to the inlet of the storage chamber 200 on the outlet side (cultured cells are placed on the inner bottom 460 for culture, and the culture medium flows through the inner bottom 460). Thus, the inner bottom 460 of the culture tank 400 and the outlet channel 450 are both on the same plane and extend upward to the inlet of the storage chamber 200. This also ensures that the culture medium in the culture tank 400 and the outlet channel 450 flows into the storage chamber 200, thereby preventing backflow.

[0067] like Figure 1 As shown, in another embodiment, the outlet channel 450 extends horizontally from the outlet of the culture tank 400 to the vertical projection position of the inlet of the storage chamber located downstream of the culture medium flow direction, and then communicates upward with the inlet. Figure 1 As shown, the liquid outlet channel 450 of the culture tank 400 first extends horizontally to the right into the interior of the liquid storage chamber 200, and then extends upward to connect with the liquid inlet.

[0068] like Figure 10As shown, in another embodiment, at least one baffle plate 451 is provided at the bottom of the outlet channel 450. The baffle plate 451 is located in the horizontal extension section of the outlet channel 450 and is inclined from the bottom of the outlet channel 450 towards the flow direction of the culture medium. Figure 10 for Figure 3 The cross-sectional view of the outlet channel 450 of the second culture tank 420 on the left side shows that during the oscillation process, when the second storage chamber 310 oscillates to a high position and the first storage chamber 210 oscillates to a low position, the culture medium flows from the second storage chamber 310 through the outlet channel 450 of the second culture tank 420 into the first storage chamber 210. If the oscillation continues until the height of the first storage chamber 210 is higher than the height of the second storage chamber 310, a portion of the culture medium may remain in the outlet channel 450, which could lead to backflow into the second culture tank 420. Therefore, a baffle plate 451 is installed at the bottom of the outlet channel 450 to block the remaining culture medium and prevent backflow into the second culture tank 420. It should be noted that in this embodiment, the baffle plate is a curved plate bent towards the first storage chamber 210. In other embodiments, it can also be a flat plate inclined towards the first storage chamber 210. This technical solution does not limit the shape of the baffle plate 451. Of course, for the first culture tank 410, the baffle plate in its liquid outlet channel is tilted toward the second liquid storage chamber to prevent the culture medium in the liquid outlet channel from flowing back into the first culture tank 410.

[0069] Continue to refer to Figure 10 In another embodiment, the at least one flow-blocking plate 451 includes a plurality of flow-blocking plates, which are spaced apart at the bottom of the liquid outlet channel along the direction from the liquid storage chamber to the culture tank, and the area of ​​the plurality of flow-blocking plates gradually decreases in the direction of liquid flow. In this embodiment, providing multiple flow-blocking plates can increase the amount of liquid blocked, for example... Figure 10 As shown, three baffles 451 with gradually increasing areas are installed between the first storage chamber 210 and the second culture tank 420. When the first storage chamber 210 swings to a higher position, the culture medium in the outlet channel 450 first converges at the first baffle. When the culture medium overflows from the first baffle, it can be blocked by the second baffle. If it continues to overflow from the second baffle, the third baffle can further block it, thus preventing the culture medium from flowing back into the second culture tank 420. The gradually increasing size of the multiple baffles 451 not only provides multiple layers of protection against backflow but also increases the maximum amount of culture medium that can be blocked.

[0070] In another embodiment, such as Figure 3 , Figure 5As shown, the inner bottom of the liquid storage chamber 200 is provided with a guide surface, and the liquid inlet is located on the guide surface or on the inner sidewall adjacent to the guide surface. The guide surface extends downward at an angle from the liquid inlet side to the liquid outlet side. Figure 5 As shown, Figure 5 for Figure 3 The image shows a right view of the first liquid storage chamber 210. The bottom of the first liquid storage chamber 210 includes a first guide surface 213 (for ease of understanding, the guide surface in the first liquid storage chamber 210 is referred to as the first guide surface 213, and the guide surface in the second liquid storage chamber 310 is referred to as the second guide surface 313). A first outlet 212 extends inward from the bottom of the outer wall of the first liquid storage chamber 210 to the inner bottom of the first liquid storage chamber 210. A first inlet 211 is located on the first guide surface 213. The first guide surface 213 slopes upward from the first outlet 212. Since the first inlet 211 is higher than the first outlet 212, during the swaying process, when the first storage chamber 210 is at a lower position, the culture medium flowing out from the second outlet 312 of the second storage chamber 310 passes through the second culture tank 420, enters through the first inlet 211, and then converges at the first outlet 212 of the first storage chamber 210 after passing through the first guide surface 213. When the first storage chamber 210 sways to a higher position, it can directly flow out from the first outlet 212 of the first storage chamber 210. In this embodiment, the inlet is located on the guide surface, for example, it can extend downwards to the height of the base 100, and then extend towards the middle culture tank 400 and communicate with the culture tank 400. In other embodiments, the inlet can also be located on the inner sidewall adjacent to the guide surface, so that the culture medium entering from the inlet can flow along the inner sidewall to the guide surface, and then converge towards the outlet.

[0071] In another embodiment, such as Figure 3 As shown, the guide surface is parallel to the line connecting the two liquid storage chambers 200. Figure 3 As shown, the inner bottom of the first liquid storage chamber 210 is provided with a first horizontal bottom surface 214 and a first guide surface 213, and the inner bottom of the second liquid storage chamber 310 is provided with a second horizontal bottom surface 314 and a second guide surface 313. Both the first guide surface 213 and the second guide surface 313 are parallel to the line connecting the first liquid storage chamber 210 and the second liquid storage chamber 310. Figure 3The base 100 swings left and right. Whether the first liquid storage chamber 210 or the second liquid storage chamber 310 swings to its lowest position, the culture medium entering from the first inlet 211 or the second inlet 311 can converge towards the first outlet 212 on the first horizontal bottom surface 214 or the second outlet 312 on the second horizontal bottom surface 314 on the opposite side. This facilitates the smooth flow of the culture medium from the outlet when the base 100 swings to its highest position. In other embodiments, the bottom of the liquid storage chamber 200 may not have a horizontal bottom surface, but only a guide surface, such as... Figure 7 As shown, taking the first liquid storage chamber 210 as an example, the inner bottom surface of the first liquid storage chamber 210 is an integrally inclined first guide surface 213, and the first liquid outlet 212 is located at the bottom of the first guide surface 213.

[0072] Continue to refer to Figure 3 In another embodiment, the inlet is located on the guide surface, and the inlet is located at the edge of the guide surface away from the culture tank 400. Taking the first storage chamber 210 on the left as an example, the first outlet 212 is connected to the first storage chamber 210 from the right side wall, and the first inlet 211 is located on the first guide surface 213 near the left side wall. So that when the first storage chamber 210 swings to a higher position, the first inlet 211 is relatively located at a higher position, further preventing the culture medium in the first storage chamber 210 from flowing out from the first inlet 211, and to a certain extent increasing the maximum amount of culture medium that the first storage chamber 210 can store. Similarly, for the second liquid storage chamber 310 located on the right, the second liquid inlet 311 is located at the edge of the second guide surface 313 near the right side wall. When the second liquid storage chamber 310 swings to a high position, the second liquid inlet 311 is in a higher position, thereby preventing the culture medium in the second liquid storage chamber 310 from flowing out from the second liquid inlet 311. This also increases the maximum amount of culture medium that the second liquid storage chamber 310 can store to a certain extent.

[0073] In another embodiment, such as Figure 8 As shown, taking the first liquid storage chamber 210 as an example, the first guide surface 213 extends upward in an inverted cone shape. For example, the inner bottom of the first liquid storage chamber 210 is funnel-shaped, and the inner bottom of the funnel shape is surrounded by a guide surface. The first outlet 212 connects from the side wall to the bottom of the funnel shape. In other embodiments, the first guide surface 213 can also be a funnel-shaped conical surface or a funnel-shaped pyramidal surface.

[0074] In another embodiment, the outlets of the two liquid storage chambers 200 are respectively located on opposite areas of the sidewalls of the two liquid storage chambers 200. For example... Figure 3As shown, the first storage chamber 210 and the second storage chamber 310 are spaced apart. The first outlet 212 and the second outlet 312 are respectively located on the inner opposite side walls of the two storage chambers 200. Thus, the outlets of the two storage chambers 200 face the other storage chamber 200, facilitating the flow of culture medium from their respective outlets during oscillation. It should be noted that when the storage chamber is cylindrical or spherical, the area where the side walls of the two storage chambers face each other is not planar. In this case, the outlets can be located in the area where the side walls of the two storage chambers are close together. In another embodiment, the two outlets can also be located at the bottom of the area where the side walls of the two storage chambers face each other. In this case, the two outlets can be directly connected to the inner bottom of their respective storage chambers 200, allowing all the culture medium to flow out from the outlets. During installation, two identical liquid storage chambers 200 can be used. First, set the first outlet 212 of one of them, such as the first liquid storage chamber 210 on the left, to the right, and move the other to the other side to a suitable distance. Then, rotate it 180° around the central axis of the liquid storage chamber 200.

[0075] like Figure 4 As shown, in another embodiment, the inner bottom of the culture tank 400 is provided with a plurality of grooves 430 spaced apart along the line connecting the two liquid storage chambers 200. The grooves 430 are used to hold cultured cells, and the height of the grooves 430 is not higher than the outlet height of the liquid inlet channel 440 within the culture tank. Figure 4 Two culture tanks 400 (first culture tank 410 and second culture tank 420) are arranged between the first culture tank 210 and the second culture tank 310. Each culture tank 400 has six recesses 430, which are arranged in a line between the first culture tank 210 and the second culture tank 310, with adjacent recesses 430 spaced apart. Each recess 430 is used to hold cultured cells. When the tank is rocked, the culture medium passes through the six recesses 430 sequentially from the left side in the first culture tank 410, and from the right side in the second culture tank 420. This arrangement can improve the efficiency of cell chip culture. On the other hand, the height of the groove 430 is not higher than the outlet height of the liquid inlet channel 440 in the culture tank. For example, if the groove 430 in the culture tank 400 is recessed from the inner bottom of the culture tank 400, then the liquid inlet channel 440 and the liquid outlet channel 450 located at both ends of the culture tank 400 can directly lead to the bottom of the culture tank 400, so that the culture medium entering the culture tank 400 can flow directly over the cultured cells in the groove 430.

[0076] like Figure 9As shown, in another embodiment, the groove 430 extends from the bottom of the culture tank 400 to the lower surface of the base 100. A light-transmitting film is provided on the lower surface of the base 100 for placing the cultured cells. By configuring the groove 430 at the bottom of the culture tank 400 for placing the cultured cells to extend through the base 100, and by providing a light-transmitting cell film on the lower side of the base 100, during use, the light-transmitting film is first attached to the lower surface of the base 100, and then the cultured cells are placed in the groove 430. During culturing, the light-transmitting film allows light to penetrate into the groove 430, thus improving cell culture.

[0077] like Figure 9 As shown, in another embodiment, the bottom surface of the base 100 is provided with a positioning groove 111, and all the grooves 430 are located within the positioning groove 111. The positioning groove 111 is used to position the light-transmitting film from the bottom of the base 100. The positioning groove 111 facilitates the application of the light-transmitting film.

[0078] In another embodiment, the base 100 can be made of a light-transmitting material, so that the entire base 100 is light-transmitting. In this case, the groove 430 of the culture tank 400 does not need to penetrate the entire base 100, and there is no need to set a light-transmitting film under the base 100. Alternatively, the light-transmitting material can be used only in the area of ​​the base 100 facing the groove 430.

[0079] like Figure 6 , Figure 9 As shown, in another embodiment, the device further includes a cover plate 600 for covering the two liquid storage chambers 200 and the culture tank 400, as illustrated. Figure 6 As shown, the cover plate 600 is located above the first liquid storage chamber 210, the second liquid storage chamber 310, and the intermediate culture tank 400, and covers the first liquid storage chamber 210, the second liquid storage chamber 310, and the intermediate culture tank 400. During the shaking process, the cover plate 600 is used to cover the liquid storage chamber 200 and the culture tank 400 to prevent dust and debris from entering.

[0080] In another embodiment, an observation window, such as a transparent observation window, can be provided in the area where the cover plate 600 covers the culture tank 400, so that staff can easily observe the culture status of the cell chip through the observation window 610.

[0081] In another embodiment, the cover plate 600 is made of a transparent material in the area covering the culture tank 400, in which case there is no need to install an observation tube.

[0082] like Figure 9As shown, in another embodiment, the cover plate 600 has a buckle on the surface of the plate covering the liquid storage chamber 200. When the cover plate 600 is closed, the two liquid storage chambers 200 have hooks (not shown in the figure) at the positions corresponding to the buckles. The use of buckles and hooks for closing facilitates disassembly, and the tight seal prevents the culture medium from overflowing during shaking.

[0083] In another embodiment, such as Figure 3 and Figure 4 As shown, the base 100 is also provided with a fixing hole 110, which is used to fix the base 100 to the rocking device. For example Figure 3 and Figure 4 As shown, three corners of the four corners of the base 100 can be selected and fixed with holes 110 to be positioned and fixed with the fixing pins on the swing device, thereby improving the stability during the swing process. Of course, in other embodiments, four or more fixing holes 110 can also be provided.

[0084] In another embodiment, the base 100 is also provided with flow direction markings. For example, in... Figure 3 The cell culture chip shown includes a first culture tank 410 and a second culture tank 420. The flow directions in the two culture tanks 420 are different. Flow direction markings can be set on the base 100 to avoid confusion. For example, one of the four corners of the base 100 can be rounded, or a line marking can be set at one end, etc.

[0085] The present invention also proposes a single-circulation fluid device, which includes the cell culture chip and shaker as described in any of the above embodiments. Since this solution incorporates the cell culture chip as described in any of the above embodiments, this device also possesses all the beneficial effects of the aforementioned cell culture chip.

[0086] In one embodiment, the cell culture chip is fixed to a shaker by a base 100. The shaker drives the cell culture chip to swing, so that the cell culture chip can continuously simulate unidirectional flow.

[0087] In one embodiment, multiple cell culture chips can be provided on the shaker, so that multiple cell culture chips can all perform unidirectional cell culture under the action of the shaker, realizing high-throughput, large-scale cell culture.

[0088] In one embodiment, the rocking bed can be a pendulum rocking bed, and in other embodiments it can be a rocker rocking bed.

[0089] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made using the contents of the present invention's specification and drawings under the inventive concept of the present invention, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention.

Claims

1. A cell culture chip, comprising a base, characterized in that, include: The base is provided with at least one pair of oppositely arranged liquid storage chambers, which are used to store culture medium. Each liquid storage chamber is provided with an outlet and an inlet. The outlet allows only the culture medium to flow out, and the inlet allows only the culture medium to flow in. At least one culture tank is provided between each pair of liquid storage chambers, which is used to culture cells. In each pair of storage chambers, the outlet of one storage chamber is connected to the inlet of the other storage chamber through the at least one culture tank, and the inlet of one storage chamber is connected to the outlet of the other storage chamber. The height of the inlet of each of the aforementioned liquid storage chambers is higher than the height of the outlet; The inner bottom of the liquid storage chamber is provided with a flow guide surface, and the liquid inlet is located on the flow guide surface or on the inner side wall adjacent to the flow guide surface. The flow guide surface extends obliquely from the side of the liquid inlet to the side of the liquid outlet. The base is also used for mounting on the swing device.

2. The cell culture chip as described in claim 1, characterized in that, Each of the liquid storage chambers is equipped with a check valve at its inlet. In each pair of storage chambers, the outlet of one storage chamber is connected to the check valve of the inlet of the other storage chamber through the at least one culture tank, and the check valve of the inlet of one storage chamber is connected to the outlet of the other storage chamber.

3. The cell culture chip as described in claim 1, characterized in that, Both ends of the culture tank are provided with an inlet channel and an outlet channel. The inlet channel is used to connect to the outlet of the storage chamber located upstream in the direction of culture medium flow, and the outlet channel is used to connect to the inlet of the storage chamber located downstream in the direction of culture medium flow.

4. The cell culture chip as described in claim 3, characterized in that, The liquid outlet channel extends upwards from the outlet of the culture tank to the inlet of the liquid storage chamber located downstream of the flow direction of the culture medium.

5. The cell culture chip as described in claim 4, characterized in that, The inner bottom of the culture tank extends at an angle from the inlet of the culture tank toward the inlet of the liquid storage chamber located on the outlet side, so that the liquid outlet channel and the inner bottom of the culture tank are on the same plane.

6. The cell culture chip as described in claim 4, characterized in that, The liquid outlet channel extends horizontally from the outlet of the culture tank to the vertical projection position of the liquid inlet of the storage chamber located downstream of the flow direction of the culture medium, and then communicates upward with the liquid inlet.

7. The cell culture chip as described in claim 6, characterized in that, At least one flow-blocking plate is provided at the bottom inner side of the liquid outlet channel. The flow-blocking plate is located in the horizontal extension section of the liquid outlet channel and is inclined from the bottom inner side of the liquid outlet channel in the direction of culture medium flow.

8. The cell culture chip as described in claim 7, characterized in that, The at least one flow-blocking plate includes multiple flow-blocking plates, which are spaced apart along the direction from the liquid storage chamber to the culture tank at the inner bottom of the liquid outlet channel, and the area of ​​the multiple flow-blocking plates gradually decreases in the direction of liquid flow.

9. The cell culture chip as described in claim 1, characterized in that, The liquid inlet is located on the guide surface, and the liquid inlet is located on the edge of the guide surface away from the culture tank.

10. The cell culture chip as described in claim 1, characterized in that, The guide surface extends upward in an inverted cone shape.

11. The cell culture chip as described in claim 1, characterized in that, In each pair of liquid storage chambers, the two liquid outlets are respectively located in the areas where the side walls of the two liquid storage chambers are opposite each other.

12. The cell culture chip as described in claim 4, characterized in that, The two liquid outlets are located at the bottom of the areas opposite each other on the side walls of the two liquid storage chambers.

13. The cell culture chip according to any one of claims 3-12, characterized in that, In each pair of storage chambers, the outlets of both storage chambers are connected to the inlet of the other storage chamber through the at least one culture tank.

14. The cell culture chip as described in claim 4, characterized in that, The bottom of the culture tank has multiple grooves spaced apart along the line connecting the two liquid storage chambers. The grooves are used to place cultured cells, and the height of the grooves is not higher than the outlet height of the liquid inlet channel in the culture tank.

15. The cell culture chip as described in claim 14, characterized in that, The groove extends from the bottom of the culture tank to the bottom surface of the base.

16. The cell culture chip as described in claim 15, characterized in that, The bottom surface of the base is provided with a positioning groove, and all the grooves are located in the positioning groove. The positioning groove is used to position the light-transmitting film from the bottom of the base.

17. The cell culture chip as described in claim 14, characterized in that, The base, including the groove, is made of a light-transmitting material within a predetermined area.

18. The cell culture chip as described in claim 1, characterized in that, The at least one culture tank includes multiple culture tanks, which are arranged side by side between the liquid outlet and the liquid inlet of the two liquid storage chambers.

19. The cell culture chip as described in claim 1, characterized in that, It also includes a cover plate for covering the two liquid storage chambers and the culture tank.

20. The cell culture chip as described in claim 19, characterized in that, The cover plate has an observation window in the area covering the culture tank.

21. The cell culture chip as described in claim 19, characterized in that, The cover plate is made of a transparent material in the area that covers the culture tank.

22. The cell culture chip according to any one of claims 19-21, characterized in that, The cover plate has a buckle on the surface of the liquid storage chamber. When the cover plate is closed, the two liquid storage chambers have hooks at the positions corresponding to the buckles.

23. The cell culture chip as described in claim 1, characterized in that, The base is also provided with fixing holes for fixing the base to the rocking device.

24. The cell culture chip as described in claim 1, characterized in that, The base is also equipped with flow direction markings.

25. A single-circulation fluid device comprising a cell culture chip as described in any one of claims 1-24, and a shaker.

26. The single-circulation fluid device as claimed in claim 25, characterized in that, The cell culture chip is fixed on the shaker via the base.

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