CN116496903A - Cell culture unit, device, method and use - Google Patents

Abstract

The application discloses a cell culture unit, a device, a method and application, wherein the cell culture unit comprises a substrate and an insert, the substrate is provided with a liquid storage cavity, a containing cavity and a runner which is communicated with the liquid storage cavity and the containing cavity; the plug-in components are located hold the chamber, and the plug-in components include main part and membrane structure, and the main part can be dismantled with the base member and be connected, and the bottom and the membrane structure of main part are connected, and the main part surrounds with the membrane structure and form first culture chamber, and first culture chamber passes through the filtration pore on the membrane structure and holds the chamber intercommunication, and the plug-in components is located and holds the chamber after, and the main part is sealed to be set up between the lateral wall in chamber with holding. In the embodiment of the application, the plug-in component is detachably connected with the matrix, so that the membrane structure can be taken out of the accommodating cavity, and the cultured organ model can be taken out more conveniently for subsequent slice analysis.

Description

Cell culture unit, device, method and use

Technical Field

The present application relates to the field of cell culture technology, and in particular, to a cell culture unit, device, method and use.

Background

The organ chip technology is a technology for in-vitro cell three-dimensional culture in a chip, and by constructing a micro-channel, a micro-reaction chamber and other functional components, components such as cells/fluid/gas/extracellular microenvironment and the like in the chip are precisely controlled, so that human body micro-tissues and micro-organs with biological functions are generated. In the related art, in order to simulate different structures in a human body, porous membranes are adopted to construct interfaces in the research of partial organ chip technology, but most porous membrane structures are integrated in the chip, and the porous membrane structures cannot be taken out, so that the subsequent analysis and experiment of a cell model are not facilitated.

Disclosure of Invention

The embodiment of the application provides a cell culture unit, a device, a method and application, which can facilitate the disassembly of a membrane structure, thereby facilitating the extraction of the membrane structure in the cell culture unit and facilitating the subsequent analysis of a cell model.

In a first aspect, embodiments of the present application provide a cell culture unit comprising a substrate and an insert, the substrate having a reservoir, a receiving chamber, and a flow channel communicating the reservoir with the receiving chamber; the plug-in unit is positioned in the accommodating cavity and comprises a main body and a membrane structure, the main body is detachably connected with the main body, the bottom end of the main body is connected with the membrane structure, the main body and the membrane structure are surrounded to form a first culture cavity, and the first culture cavity is communicated with the accommodating cavity through a filter hole on the membrane structure; after the plug-in components are arranged in holding the chamber, sealed setting is between the lateral wall in main part and holding the chamber.

In some embodiments of the present application, the insert comprises a body, the base further having a second culture chamber disposed in the flow channel, and the second culture chamber is located below the membrane structure.

In some embodiments of the present application, the liquid storage cavities are disposed on two opposite sides of the accommodating cavity, and the liquid storage cavities located on two sides of the accommodating cavity are communicated with the accommodating cavity through the flow channel.

In some embodiments of the present application, the orthographic projection of the flow channel on the horizontal plane is in a fusiform shape, the second culture cavity is located in the middle of the flow channel, the accommodating cavity is communicated with the middle of the flow channel, and the liquid storage cavities located on two sides of the accommodating cavity are respectively communicated with two ends of the flow channel.

In some embodiments of the present application, the substrate includes a liquid storage layer and a bottom plate, where the liquid storage cavity, the accommodating cavity and the flow channel are all disposed on the liquid storage layer, and the flow channel is located at the bottom of the accommodating cavity; the bottom plate is connected with the bottom of the liquid storage layer, and seals the bottom of the liquid storage cavity and the flow channel.

In some embodiments of the present application, the flow channel extends along a first preset direction to communicate the liquid storage cavity with the accommodating cavity; along a second preset direction, the orthographic projection width of the liquid storage cavity on the horizontal plane is larger than the orthographic projection width of the flow channel on the horizontal plane, the second preset direction is perpendicular to the first preset direction, and the second preset direction is parallel to the horizontal plane.

In some embodiments of the present application, a first opening communicating with the accommodating cavity is formed in the base body, the first opening is used for allowing the insert to enter and exit the accommodating cavity, and a first step structure is arranged on an inner side wall of the first opening; the end part of the plug-in is provided with a second step structure, the second step structure and the first step structure are distributed along the direction that the plug-in is inserted into the accommodating cavity, and after the plug-in is positioned in the accommodating cavity, the second step structure is abutted with the first step structure.

In some embodiments of the present application, the substrate has a first top surface, the first opening is formed on the first top surface, the second step structure has a second top surface, and after the insert is located in the accommodating cavity, the first top surface and the second top surface are located in the same plane.

In some embodiments of the present application, a first step structure is disposed around a perimeter side of an inner sidewall of the first opening, and a second step structure is disposed around a perimeter side of the insert end; and/or the first step structure has a recess, and the second step structure has a protrusion that mates with the recess.

In a second aspect, an embodiment of the present application further provides a cell culture apparatus, including a culture plate and a plurality of cell culture units as described in any one of the above embodiments, where the plurality of cell culture units are arranged in an array on the culture plate.

In a third aspect, an embodiment of the present application further provides a cell culture method, which is applied to the cell culture apparatus described in the above embodiment, the cell culture method including the steps of:

injecting a biological sample into a first culture chamber of the cell culture unit;

adding a culture medium into a liquid storage cavity of the cell culture unit, and enabling the culture medium to infiltrate the biological sample;

and performing swing adjustment on the cell culture device to dynamically culture the biological sample to obtain a three-dimensional cell spheroid cell model.

In a fourth aspect, the present application also provides the use of a cell culture apparatus in biological model culture and drug analysis.

The beneficial effects of the embodiment of the application are that: the cells are cultured in the cell culture unit to construct an organ model, and the insert and the matrix are detachably connected in the embodiment of the application, so that the membrane structure can be taken out of the accommodating cavity, and the cultured organ model can be taken out more conveniently for subsequent slice analysis; meanwhile, the detachable plug-in components also enable complete co-culture detection of different organs to be more convenient;

in addition, the side wall of the accommodating cavity and the main body are arranged in a sealing manner, and culture mediums in the liquid storage cavity and the flow channel cannot run off through gaps between the side wall of the accommodating cavity and the main body, so that the cell culture unit can perform inverted culture, namely cells can be attached to the bottom wall of the membrane structure, and therefore the upper surface and the lower surface of the membrane structure can perform cell culture operation.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic diagram of an exploded structure of a cell culture unit according to an embodiment of the present application;

FIG. 2 is a schematic diagram showing an inverted structure of a cell culture unit according to an embodiment of the present application;

FIG. 3 is a schematic diagram showing an inverted structure of a cell culture unit according to another embodiment of the present application;

FIG. 4 is a schematic cross-sectional view of a cell culture unit according to an embodiment of the present disclosure;

FIG. 5 is a schematic diagram showing the structure of a cell culture unit according to another embodiment of the present application;

FIG. 6 is a schematic diagram of a cell culture apparatus according to an embodiment of the present disclosure;

FIG. 7 is a flow chart of a cell culture method according to an embodiment of the present application.

Reference numerals:

1. a cell culture unit; 10. a base; 11. a liquid storage cavity; 12. a receiving chamber; 13. a flow passage; 14. a liquid storage layer; 15. a bottom plate; 16. a first opening; 161. a first step structure; 1611. a concave portion; 17. a second culture chamber; 18. a first top surface; 20. an insert; 21. a first culture chamber; 22. a membrane structure; 23. a main body; 24. a second step structure; 241. a convex portion; 242. a second top surface; 2. a cell culture device; 201. a culture plate; l1, a first preset direction; l2, a second preset direction; l3, a third preset direction; and L4, a fourth preset direction.

Detailed Description

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the related art, the following description will be made in detail with reference to the accompanying drawings in the embodiments of the present application, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

In the related art, in membrane-type organ chips, porous membranes are mostly integrated in the chip, cannot be taken out, are unfavorable for subsequent analysis of a model, and are unfavorable for the integration of a single chip into a chip system to construct a multi-organ chip.

In view of the above, referring to fig. 1, the present application proposes a cell culture unit 1, including a substrate 10 and an insert 20, wherein the substrate 10 has a liquid storage chamber 11, a receiving chamber 12, and a flow channel 13 communicating the liquid storage chamber 11 and the receiving chamber 12. Wherein, the culture medium can be added in the liquid storage cavity 11, and the culture medium in the liquid storage cavity 11 can enter the containing cavity 12 through the flow channel 13.

The insert 20 is located and holds the chamber, and the insert 20 includes main part 23 and membrane structure 22, and main part 23 and base member detachable connection, and the bottom of main part 23 is connected with membrane structure 22, and main part 23 and membrane structure 22 surround and form first culture chamber 21, and first culture chamber 21 passes through the filtration pore on the membrane structure 22 and holds chamber 12 intercommunication.

Specifically, a mixed culture medium containing a biological sample may be added to the first culture chamber 21, and the culture medium added to the liquid storage chamber 11 may first enter the accommodating chamber 12 and then enter the first culture chamber 21 through the filter holes of the membrane structure 22. In this way, the staff can add the culture medium mixture containing the biological sample into the first culture cavity 21 of the insert 20, and add the culture medium, the specific medicine or the culture medium of the specific medicine into the liquid storage cavity 11, so that the culture medium in the liquid storage cavity 11 can provide nutrition for the biological sample in the first culture cavity 21, and the medicine can act on the biological sample. The membrane structure 22 may be made of plastic material such as PC or PET, so as to ensure that the membrane structure 22 can allow the culture medium in the accommodating chamber 12 to pass through the filter hole and enter the first culture chamber 21. In addition, the main body 23 of the insert 20 and the film structure 22 may be bonded and packaged by hot pressing, ultrasonic wave, laser, etc. to ensure the connection firmness between the film structure 22 and the main body 23.

After the insert 20 is positioned in the receiving chamber 12, the body 23 is sealingly disposed between the side walls of the receiving chamber 12. It will be appreciated that after the insert 20 is placed in the accommodating cavity 12, a sealed space is formed between the accommodating cavity 12 and the main body 23, and when the cell culture unit 1 is cultured upside down, the culture medium in the liquid storage cavity 11 and the flow channel 13 cannot be lost through the gap between the accommodating cavity 12 and the main body 23, so that the waste of the culture medium is reduced, and meanwhile, the cell culture unit 1 can be cultured upside down, that is, cells can be attached to the bottom wall of the membrane structure 22, so that the cell culture operation can be performed on both the upper surface and the lower surface of the membrane structure 22. The sealing method between the main body 23 and the accommodating cavity 12 is not particularly limited, for example, the shape of the main body 23 and the shape of the accommodating cavity 12 may be set to match the two to achieve the sealing effect between the main body 23 and the accommodating cavity 12 after the insert 20 is inserted into the accommodating cavity 12; alternatively, a sealing ring is provided between the body 23 and the accommodating chamber 12 so that the body 23 is sealed with the accommodating chamber 12.

It should be noted that, by performing cell culture on the membrane structure 22 in the first culture chamber 21, a three-dimensional organ model, such as various biological barrier models of blood brain barrier, kidney barrier, intestine barrier, lung barrier, skin barrier, etc., may be constructed. In the embodiment of the application, the insert 20 is detachably arranged in the accommodating cavity 12 of the substrate 10, so that the membrane structure 22 can be taken out of the accommodating cavity 12, and the cultured organ model can be taken out of the cell culture unit 1 conveniently for separate subsequent slice analysis and the like.

Meanwhile, in the related art, the culture mediums of different kinds of organs are different, for example, the culture mediums of a first organ may cause damage to a second organ, but in the embodiment of the present application, after the different kinds of organs are individually cultured, the organ model which has been cultured may be individually taken out, and then placed under the condition of co-culture, so that short-time detection is performed on the different kinds of organ models, whereby the detachable membrane structure provides convenience for the operation. That is, in the related art, the porous membrane structure 22 in the cell culture unit 1 is not detachable, and in the technical background that co-culture cannot be achieved by different types of organ models, it may cause inconvenience in experiments and increase in experimental costs. In the embodiment of the present application, the insert 20 may be removed from the substrate 10, which is also more beneficial for integrating a cultured organ model into other different organ systems, so as to facilitate drug detection on multiple different organ models, thereby greatly improving the detection efficiency of the drug on the organ.

It should be further noted that the membrane structure 22 is located at the bottom end of the main body 23, so that after the cell culture unit 1 is left for a period of time, cells in the culture medium can be deposited on the bottom of the main body 23 and adsorbed on the upper surface of the membrane structure 22, thereby facilitating the growth of the cultured cells to construct an organ model. After the culture medium mixed solution containing the biological sample is injected into the first culture cavity 21, the culture medium mixed solution can flow under the action of self gravity, so that a fluid shear force environment is provided for the growth of cells in the culture medium, a fluid driving mode without pump type gravity driving is simpler, and the construction cost is lower. Of course, the mixed culture solution containing biological samples, the culture medium and the medicines can be added in a pump-free gravity driving mode, and an external peristaltic pump, a syringe pump and other devices can be used for driving the culture medium to flow so as to provide a growth environment required by cells. In addition, the shapes of the liquid storage chamber 11, the accommodating chamber 12 and the first culturing chamber 21 are not particularly limited, and the shapes of the liquid storage chamber 11, the accommodating chamber 12 and the first culturing chamber 21 may be the same or different, for example, the liquid storage chamber 11 and/or the first culturing chamber 21 may be irregularly shaped, or the cross section of the liquid storage chamber 11 may be square, and the cross section of the accommodating chamber 12 and the first culturing chamber 21 may be circular.

Referring to FIGS. 1-2, in some embodiments of the present application, the flow channel 13 has a second culture chamber 17 therein, and the second culture chamber 17 is located below the membrane structure 22.

It should be noted that, when the culture medium containing the biological sample is injected into the liquid storage cavity 11 and the flow channel 13, and then the cell culture unit 1 is cultured upside down, the culture medium will culture the cells in the second culture cavity 17 of the flow channel 13, and the second culture cavity 17 is located below the membrane structure 22, so that the cells can be attached to the bottom wall of the membrane structure 22, and thus, the purpose that the cell culture operation can be performed on both the upper surface and the lower surface of the membrane structure 22 is achieved.

With continued reference to fig. 1-2, in some embodiments of the present application, the storage chambers 11 are disposed on opposite sides of the accommodating chamber 12, and the storage chambers 11 disposed on opposite sides of the accommodating chamber 12 are in communication with the accommodating chamber 12 through the flow channels 13.

It will be appreciated that medium is added to the reservoir 11 on one side of the receiving chamber 12 and enters the reservoir 11 on the other side of the receiving chamber 12 through the flow channel 13; alternatively, the same culture medium is simultaneously added into the liquid storage cavities 11 at both sides of the accommodating cavity 12, and the culture medium can flow through the flow channel 13; or, different culture mediums are respectively added into the liquid storage cavities 11 at two sides of the accommodating cavity 12, and the culture mediums arranged in the two liquid storage cavities 11 can exchange substances with biological samples so as to meet the requirement of cell growth in the culture mediums on nutrient substances.

It should be noted that, when the cell culture unit 1 is reciprocally swung through a specific angle, the culture medium reciprocally flows between the liquid storage chambers 11, the flow channels 13 and the accommodating chambers 12 located at both sides of the accommodating chambers 12, and the continuously flowing culture medium can dynamically culture the cells on the upper surface of the membrane structure 22 through the filtering holes on the membrane structure 22, and meanwhile, the waste generated by the biological sample in the culture chamber 21 can be taken away, so that the real cell growth environment can be more simulated. The number of the liquid storage cavities 11 is not particularly limited, and the number of the liquid storage cavities 11 may be 3, 4 or 5 according to the specific requirement of the culture.

In some embodiments of the present application, referring to fig. 2, the orthographic projection of the flow channel 13 on the horizontal plane is in a fusiform shape, the second culture cavity 17 is located in the middle of the flow channel 13, the accommodating cavity 12 is communicated with the middle of the flow channel 13, and the liquid storage cavities 11 located at two sides of the accommodating cavity 12 are respectively communicated with two ends of the flow channel 13.

It should be noted that, if the front projection of the flow channel 13 on the horizontal plane is in the form of a shuttle, the end of the flow channel 13 is narrower, the flow rate of the culture medium flowing into the flow channel 13 from the liquid storage chamber 11 is smaller, the flow rate of the culture medium in the flow channel 13 is reduced, so that the time for the culture medium and the medicine to flow through the flow channel 13 is longer, the culture medium can be contacted with the biological sample in the culture chamber 21 for a longer time, and the middle of the flow channel 13 is wider, so that more culture medium can enter the accommodating chamber 12 and enter the culture chamber 21 through the membrane structure 22.

It should be further noted that, the outlets at both ends of the shuttle-shaped flow channel 13 are narrower, the middle space of the shuttle-shaped flow channel 13 is larger, and the second culture chamber 17 is located at the middle of the flow channel 13, when the cell culture unit 1 is inverted, the surface tension of the liquid formed by the culture medium in the flow channel 13 is larger when the culture medium is cultured in the second culture chamber 17, and the culture medium can flow out of the flow channel 13 by means of the surface tension of the liquid, so that the cells can be attached to the bottom wall of the membrane structure 22 and normally cultured in the second culture chamber 17.

In some embodiments of the present application, referring to fig. 1-3, a substrate 10 includes a liquid storage layer 14 and a bottom plate 15, where the liquid storage chamber 11, the accommodating chamber 12 and the flow channel 13 are all disposed on the liquid storage layer 14, and the flow channel 13 is located at the bottom of the accommodating chamber 12; the bottom plate 15 is connected to the bottom of the liquid storage layer 14, and seals the bottom of the liquid storage chamber 11 and the flow channel 13.

It will be appreciated that the flow channel 13 is disposed on the lower surface of the liquid storage layer 14, and is connected to the liquid storage layer through a bottom plate, so as to achieve the purpose of sealing the flow channel, and of course, the flow channel 13 may also be disposed on one side of the bottom plate 15 near the liquid storage layer 14. The substrate 10 may be composed of a liquid storage layer 14 and a bottom plate 15, the liquid storage layer 14 is covered on the bottom plate 15, and the preparation materials of the liquid storage layer 14 and the bottom plate 15 may be glass, plastic, PDMS (Polydimethylsiloxane) or the like, so that the cell culture unit 1 can have good biocompatibility by reasonably selecting the preparation materials of the liquid storage layer 14 and the bottom plate 15. Meanwhile, the connection mode between the liquid storage layer 14 and the bottom plate 15 may be hot pressing, ultrasonic wave, laser, etc., or the liquid storage layer 14 and the bottom plate 15 may be integrally formed by a 3D printing technology.

It should be noted that, the flow channel 13 is located below the accommodating cavity 12, and the membrane structure 22 is located at the bottom end of the main body 23, so that the culture medium is more convenient for culturing the cells located on the upper surface of the membrane structure 22, and the influence of the drug on the cells is facilitated to be studied.

Referring to fig. 3-4, in other embodiments of the present application, the flow channel 13 extends along the first preset direction L1 to communicate the liquid storage cavity 11 with the accommodating cavity 12; along a second preset direction L2, the orthographic projection width of the liquid storage cavity 11 on the horizontal plane is larger than the orthographic projection width of the flow channel 13 on the horizontal plane, the second preset direction L2 is perpendicular to the first preset direction L1, and the second preset direction L2 is parallel to the horizontal plane.

Similarly, with the front projection width of the liquid storage chamber 11 on the horizontal plane being H1, the front projection width of the flow channel 13 on the horizontal plane being H2, the width H1 of the liquid storage chamber 11 along the second preset direction L2 is larger than the width H2 of the flow channel 13 along the second preset direction L2, that is, H1> H2, the width from the accommodating chamber 12 to the flow channel 13 is reduced, that is, the entrance of the culture medium into the flow channel 13 from the liquid storage chamber 11 is narrowed, so that the flow velocity of the culture medium in the flow channel 13 can be reduced, and the time for the culture medium and the medicine to flow through the flow channel 13 can be prolonged, thereby the culture medium can be contacted with the biological sample in the first culture chamber 21 for a longer time.

Referring to fig. 4-5, in some embodiments of the present application, a first opening 16 communicating with the accommodating cavity 12 is formed in the base 10, the first opening 16 is used for the insert 20 to enter and exit the accommodating cavity 12, and a first step structure 161 is disposed on an inner sidewall of the first opening 16; the end of the insert 20 is provided with a second step structure 24, the second step structure 24 and the first step structure 161 are arranged along the direction that the insert 20 is inserted into the accommodating cavity 12, and after the insert 20 is positioned in the accommodating cavity 12, the second step structure 24 is abutted with the first step structure 161.

It should be noted that, taking the direction in which the insert 20 is inserted into the accommodating cavity 12 as the third preset direction L3 and the direction in which the insert 20 is extracted from the accommodating cavity 12 as the fourth preset direction L4 as an example, the first step structure 161 and the second step structure 24 are arranged along the third preset direction L3, when the insert 20 moves toward the third preset direction L3, the insert 20 is inserted into the accommodating cavity 12 through the first opening 16 on the base 10 until the second step structure 24 on the end of the insert 20 abuts against the first step structure 161 on the inner sidewall of the first opening 16, the first step structure 161 blocks the insert 20 from continuing to move along the third preset direction L3, that is, the first step structure 161 blocks the insert 20 from continuing to extend into the accommodating cavity 12, and at this time the insert 20 is fixed in the accommodating cavity 12; when it is necessary to remove the card 20 from the housing cavity 12, the card 20 is moved in the fourth preset direction L4, and the second step structure 24 at the end of the card 20 is separated from the first step structure 161.

In some embodiments of the present application, referring to fig. 4-5, the substrate 10 has a first top surface 18, the first opening 16 is formed on the first top surface 18, the second step structure 24 has a second top surface 242, and after the insert 20 is located in the accommodating cavity 12, the first top surface 18 and the second top surface 242 are located in the same plane.

It will be appreciated that the cell culture unit 1 may have an upper cover (not shown) that covers the base 10, and that the first top surface 18 is flush with the second top surface 242 when the insert 20 is placed in the accommodating chamber 12, i.e. the first top surface 18 and the second top surface 242 are in the same plane, and that no interference occurs between the insert 20 and the upper cover, so that the liquid storage chamber 11 is sealed from the accommodating chamber 12, and a good sealing environment is provided for cell culture.

Further, referring to fig. 1 and 4, in some embodiments of the present application, a first step structure 161 is disposed around the perimeter of the inner sidewall of the first opening 16 and a second step structure 24 is disposed around the perimeter of the end of the insert 20.

It will be appreciated that the first step structure 161 is located circumferentially around the inner side wall of the first opening 16 and the second step structure 24 is located circumferentially around the end of the insert 20, thereby providing a greater contact area between the first step structure 161 and the second step structure 24 and a more stable placement of the insert 20.

Referring to fig. 5, in other embodiments of the present application, the first step structure 161 has a concave portion 1611, and the second step structure 24 has a convex portion 241 that mates with the concave portion 1611.

In particular, the provision of the boss 241 may facilitate the handling of a worker or machine, making it easier to remove the insert 20 from the receiving cavity 12. Meanwhile, when the first step structure 161 abuts against the second step structure 24, the concave portion 1611 cooperates with the convex portion 241, so that the insert 20 can be prevented from shaking or rotating in the accommodating chamber 12,

it should be noted that, as shown in fig. 5, the first step structure 161 may be disposed around the inner side wall of the first opening 16 and have a concave portion 1611, and the second step structure 24 may be disposed around the end portion of the insert 20 and have a convex portion 241, so that the insert 20 not only provides a more stable culture environment for the biological sample in the first culture chamber 21, but also facilitates the taking of the worker or the machine.

In a second aspect, referring to fig. 6, an embodiment of the present application further provides a cell culture apparatus 2, including a culture plate 201 and a plurality of cell culture units 1 as described in any of the above embodiments, where the plurality of cell culture units 1 are arranged in an array on the culture plate 201.

Specifically, in the same row or the same column, the spacing between two adjacent cell culture units 1 is the same, so that an automatic pipetting machine is convenient to add culture medium to the cell culture units 1, and the spacing between two adjacent cell culture units 1 in the embodiment of the present application is not limited, and may be, for example, 9 mm, 10 mm, 12 mm, etc. Wherein, be provided with a plurality of mounting grooves that are the array arrangement on the culture plate 201, each cell culture unit 1 is placed in a mounting groove, and the mounting groove provides installation space and support for cell culture unit 1.

It should be noted that the plurality of cell culture units 1 may be arranged in M columns by N rows, where M represents the number of cell culture units 1 in each row, N represents the number of cell culture units 1 in each column, M is greater than or equal to 1, N is greater than or equal to 1, M and N are integers, and M and N are not 1 at the same time. The number of the cell culture units 1 in the cell culture apparatus 2 is not particularly limited in this embodiment, for example, the number of the cell culture units 1 may be 32, 64, 80 or 96, etc., so as to achieve the requirement of the cell culture apparatus 2 for high throughput, thereby facilitating large-scale screening of drugs. As shown in fig. 6, the cell culture apparatus 2 includes 32 cell culture units 1, and the 32 cell culture units 1 are arranged in 8 rows by 4 columns.

It should be further noted that, the cells are cultured in the cell culture unit 1 to construct an organ model, and the detachable connection of the insert 20 and the substrate 10 also makes it easier to take out the organ model for slice analysis, and the detachable insert 20 also makes it easier to complete co-culture detection of different types of organs. Meanwhile, the cell culture unit 1 can be driven by fluid driven mode of pump-free gravity driving, and the cell culture device 2 is provided with the cell culture unit 1 with high flux, so that not only can the complexity of the cell culture device 2 be reduced, but also the construction cost of a cell model can be reduced, and the cost of manpower, material resources and time for constructing a biological model and developing new drugs is greatly reduced.

In a third aspect, referring to fig. 7, an embodiment of the present application further provides a cell culture method applied to the cell culture apparatus 2 described in the above embodiment, where the cell culture method includes the following steps:

s101, adding a first preset volume of the culture medium mixed solution containing the biological sample into the first culture cavity 21 of the cell culture unit 1, and injecting a second preset volume of the culture medium into the liquid storage cavity 11.

Specifically, the injection may be performed manually or automatically by a machine for automated pipetting.

S102, placing the injected cell culture unit 1 in an incubator at a preset temperature for static culture for a preset time.

S103, placing the cell culture apparatus 2 on the swing device and performing reciprocating swing at a preset angle so that the culture medium flows back and forth between the liquid storage chamber 11 and the accommodating chamber 12.

Specifically, the swinging equipment can be a swinging table, and the swinging table can drive the cell culture device 2 to swing reciprocally by a preset angle only by setting the swinging preset angle of the swinging table, so that the cell culture device 2 does not need to be manually moved, and the efficiency of the cell culture device 2 is higher.

And S104, periodically replacing the culture medium in the liquid storage cavity 11, and finally obtaining a cell model.

In the above cell culture method, taking skin epithelial cells as an example, a first preset volume of the culture medium mixture containing epithelial cells is injected into the first culture chamber 21 of the insert 20 by the pipette gun, and a second preset volume of the culture medium is injected into the liquid storage chamber 11; then placing the cell culture unit 1 into an incubator at a preset temperature for stationary culture for a preset time, wherein the epithelial cells are deposited and adsorbed on the upper surface of the membrane structure 22; the culture medium above the membrane structure 22 is then aspirated again by a pipette (the culture of the skin needs to be exposed to air) so that the skin epithelial cells adsorbed on the upper surface of the membrane structure 22 can be exposed to air, thereby simulating the growth environment of normal skin; then placing the cell culture device 2 on a shaking table, and oscillating the shaking table through a preset angle to enable the culture medium to flow back and forth between the liquid storage cavity 11 and the accommodating cavity 12, wherein the continuously and reciprocally flowing culture medium forms dynamic culture on the skin epithelial cells through the membrane structure 22; a specific drug or a drug-containing medium or the like may then be periodically added to the reservoir 11 to study the effect of the drug on the skin epithelial cells.

In a fourth aspect, the present application also provides the use of the cell culture apparatus 2 in biological model culture and drug analysis.

The same or similar reference numerals in the drawings of the present embodiment correspond to the same or similar components; in the description of the present application, it should be understood that, if there is an azimuth or positional relationship indicated by terms such as "upper", "lower", "left", "right", etc., based on the azimuth or positional relationship shown in the drawings, this is for convenience of description and simplification of the description, but does not indicate or imply that the components or elements referred to must have a specific azimuth, be configured and operated in a specific azimuth, and thus the terms describing the positional relationship in the drawings are merely for exemplary illustration, and should not be construed as limitations of the present application, and specific meanings of the terms described above may be understood by those of ordinary skill in the art according to specific circumstances.

The foregoing description of the preferred embodiment of the present invention is not intended to limit the invention to the particular form disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Claims (12)

1. A cell culture unit comprising:

the base body is provided with a liquid storage cavity, a containing cavity and a flow passage communicated with the liquid storage cavity and the containing cavity;

the plug-in components are located hold the chamber, the plug-in components include main part and membrane structure, the main part with the base member is dismantled and is connected, the bottom of main part with membrane structure is connected, just the main part with the membrane structure surrounds and forms first cultivates the chamber, first cultivates the chamber through the filtration pore on the membrane structure with hold the chamber intercommunication, the plug-in components are located hold the chamber after, the main part with hold sealed setting between the lateral wall in chamber.

2. The cell culture unit of claim 1, wherein the substrate further has a second culture chamber disposed in the flow channel, and the second culture chamber is located below the membrane structure.

3. The cell culture unit of claim 2, wherein the reservoir chambers are provided on opposite sides of the receiving chamber, and wherein the reservoir chambers on both sides of the receiving chamber are in communication with the receiving chamber through the flow channel.

4. A cell culture unit according to claim 3 wherein the orthographic projection of the flow channel on a horizontal plane is in the shape of a shuttle, the second culture chamber is positioned in the middle of the flow channel, the accommodating chamber is communicated with the middle of the flow channel, and the liquid storage chambers positioned at both sides of the accommodating chamber are respectively communicated with both ends of the flow channel.

5. The cell culture unit of claim 1, wherein the matrix comprises:

the liquid storage layer, the liquid storage cavity, the accommodating cavity and the flow channel are all arranged on the liquid storage layer, and the flow channel is positioned at the bottom of the accommodating cavity;

and the bottom plate is connected with the bottom of the liquid storage layer and seals the bottom of the liquid storage cavity and the flow channel.

6. The cell culture unit of claim 1, wherein the flow channel extends in a first predetermined direction to communicate the reservoir with the receiving chamber;

along a second preset direction, the orthographic projection width of the liquid storage cavity on the horizontal plane is larger than the orthographic projection width of the flow channel on the horizontal plane, the second preset direction is perpendicular to the first preset direction, and the second preset direction is parallel to the horizontal plane.

7. The cell culture unit of claim 1, wherein the base body is provided with a first opening communicated with the accommodating cavity, the first opening is used for allowing the plug-in to enter and exit the accommodating cavity, and the inner side wall of the first opening is provided with a first step structure;

the end part of the plug-in is provided with a second step structure, the second step structure and the first step structure are distributed along the direction that the plug-in is inserted into the accommodating cavity, and after the plug-in is positioned in the accommodating cavity, the second step structure is abutted to the first step structure.

8. The cell culture unit of claim 7, wherein the base has a first top surface, the first opening is open on the first top surface, the second step structure has a second top surface, and the first top surface is in the same plane as the second top surface after the insert is positioned in the receiving cavity.

9. The cell culture unit of claim 7, wherein the first step structure is disposed around a perimeter of an inner sidewall of the first opening and the second step structure is disposed around a perimeter of the insert end; and/or the number of the groups of groups,

the first step structure has a recess, and the second step structure has a protrusion that mates with the recess.

10. A cell culture apparatus comprising a culture plate and a plurality of cell culture units according to any one of claims 1 to 9, wherein a plurality of the cell culture units are arranged in an array on the culture plate.

11. A cell culture method, characterized by being applied to the cell culture apparatus as claimed in claim 10, comprising the steps of:

injecting a biological sample into a first culture chamber of the cell culture unit;

adding a culture medium into a liquid storage cavity of the cell culture unit, and enabling the culture medium to infiltrate the biological sample;

and performing swing adjustment on the cell culture device to dynamically culture the biological sample to obtain a three-dimensional cell spheroid cell model.

12. Use of the cell culture apparatus of claim 10 in biological model culture and pharmaceutical analysis.