CN214612545U - Detachable/assemblable co-culture organ chip - Google Patents

Abstract

The application relates to the technical field of biological tissue engineering, and discloses a detachable/assemblable co-culture organ chip which comprises a chip platform, wherein one or more central liquid storage holes are formed in the chip platform, the central liquid storage holes are in a step shape, mounting positions are arranged on the bottom walls of the central liquid storage holes, and co-culture channels are axially arranged on the step surfaces of the central liquid storage holes in an extending mode; cultivate the cell module, detachably sets up in the mounted position department of the diapire in central stock solution hole, cultivates and is provided with one or more cultivation holes on the cell module. The co-culture organ chip of the embodiment of the disclosure is in an open type, and the openings of the culture holes and the co-culture channel face to the same side, thereby greatly reducing the operation difficulty. The region that contains the culture hole on the diapire with central stock solution hole is cut apart, forms and cultivates the cell module, makes this region can be replaced by in a flexible way to can reach accurate experimental design's requirement, it is nimble changeable, increase the application scene of organ chip, improve the application upper limit of organ chip.

Description

Detachable/assemblable co-culture organ chip

Technical Field

The present application relates to the field of biological tissue engineering, for example to a detachable/assemblable co-culture organ chip.

Background

Cells or tissues require interaction and communication between organs within the intact body, such as innervation of the nervous system, "dialogues" of the immune system, nutritional support of mesenchymal cells. At present, although traditional well plate culture platforms such as 24, 96, 384 well plates are widely applied in biological research fields such as cell culture, there are some technical bottlenecks that limit the application of the traditional well plate culture platforms in organ co-culture. The common culture well plate can not realize the intercellular coculture. The microfluidic organ chip technology is a new technology, and can be applied to co-culture of various organs by designing various channels to overcome the limitation. However, there are some technical bottlenecks to limit the large-scale application of the organ chip technology reported at present. Firstly, the stability and reproducibility of the product are poor, the product structure is complex, if a closed channel is formed, the operation of a special technician is needed, the stability and reproducibility are poor, and the subsequent analysis difficulty is high. Secondly, the flux of the product is low, the standardization degree is poor, the compatibility of the equipment is poor, and the popularization and the application are difficult.

In the process of implementing the embodiments of the present disclosure, it is found that at least the following problems exist in the related art: the existing co-culture organ chip is of a closed structure, and the structure of the chip is fixed and cannot be flexibly changed.

SUMMERY OF THE UTILITY MODEL

The following presents a simplified summary in order to provide a basic understanding of some aspects of the disclosed embodiments. This summary is not an extensive overview nor is intended to identify key/critical elements or to delineate the scope of such embodiments but rather as a prelude to the more detailed description that is presented later.

The disclosed embodiments provide an assembled co-culture organ chip to solve the problems of the prior co-culture organ chip being of a closed structure and the chip structure being fixed.

In some embodiments, the assembled co-cultured organ chip comprises: the chip platform is provided with one or more central liquid storage holes, the central liquid storage holes are in a step shape, the bottom wall of each central liquid storage hole is provided with a mounting position, and the step surface of each central liquid storage hole is axially extended and provided with a co-culture channel; cultivate the cell module, detachably sets up in the mounted position department of the diapire in central stock solution hole, cultivates and is provided with one or more cultivation holes on the cell module.

The assembled co-culture organ chip provided by the embodiment of the disclosure can realize the following technical effects:

the detachable/assemblable co-culture organ chip provided by the embodiment of the disclosure is open, the openings of the culture partition and the co-culture channel face to the same side, and cell planting can be completed on the same side (e.g., upper side) of the organ chip, thereby greatly reducing the operation difficulty. Moreover, on the basis of the integrally designed co-culture organ chip, the region containing the culture partition on the bottom wall of the central liquid storage hole is divided to form a culture chamber module, so that the region can be flexibly replaced. Therefore, the culture chamber modules with proper shapes, numbers and sizes of the culture partitions can be selected to be assembled on the chip platform according to actual requirements, so that the requirement of accurate test design can be met, flexibility and changeability are realized, the application scenes of the organ chips are increased, and the application upper limit of the organ chips is improved. Furthermore, by carrying out standardized design on the arrangement layout of the central liquid storage hole on the chip platform, the open type operation channel has strong compatibility with commercial operation, detection equipment and imaging equipment, is convenient for subsequent on-board detection and cell recovery to carry out analysis such as RNA and protein extraction, is simpler and more suitable for industrial popularization. The method is simple to operate, does not need professional technicians, enlarges the application range of the organ chip and improves the universality. The detachable/assemblable co-culture organ chip can realize contact type/non-contact type co-culture of cell or tissue micro-organs, and can realize interaction on the premise of mutual non-pollution. The method can be used for in vitro construction and long-time culture of multi-cell and multi-organ models, and further can be used for in vitro construction of multi-cell co-culture models or multi-organ co-culture models.

The foregoing general description and the following description are exemplary and explanatory only and are not restrictive of the application.

Drawings

One or more embodiments are illustrated by way of example in the accompanying drawings, which correspond to the accompanying drawings and not in limitation thereof, in which elements having the same reference numeral designations are shown as like elements and not in limitation thereof, and wherein:

FIG. 1 is a schematic top view of a culture unit of an assembled co-culture organ chip according to an embodiment of the present disclosure;

FIG. 2 is an exploded view of a cross-sectional structure taken along line A-A of FIG. 1;

FIG. 3 is a schematic diagram of a culture chamber module according to an embodiment of the present disclosure;

FIG. 4 is a schematic structural view of another culture chamber module provided in the embodiments of the present disclosure;

FIG. 5 is a schematic diagram of another culture chamber module provided in embodiments of the present disclosure;

FIG. 6 is a schematic top view of another culture unit of an assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 7 is a schematic top view of another culture unit of an assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 8 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 9 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 10 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 11 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 12 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 13 is an exploded view of another alternative culture unit of an assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 14 is a schematic top view showing a structure of a culture unit of another open-type co-culture organ chip according to the embodiment of the present disclosure;

FIG. 15 is a schematic cross-sectional view taken along line D-D of FIG. 14;

FIG. 16 is a schematic cross-sectional view of a culture unit of another assembled co-culture organ chip according to the embodiment of the present disclosure;

FIG. 17 is a schematic diagram of an assembled co-culture organ chip according to an embodiment of the present disclosure;

reference numerals:

10. a central liquid storage hole; 11. a step surface; 12. a large pore diameter section; 13. a small bore section; 20. a culture well; 30. a co-culture channel; 301. an upper liquid storage section; 302. a lower layer culture section; 40. a lateral liquid storage hole; 41. a first side liquid storage hole; 42. a second side liquid storage hole; 50. a communication channel; 51. a first communicating passage; 52. a second communicating passage; 100. a chip platform; 101. fencing; 102. a liquid containing groove; 110. an upper stage; 120. a lower stage; 150. a central liquid storage straight hole; 200. a culture chamber module; 210. a culture area; 211. a separating edge; 220. a wall; 300. a culture channel module; 310. an annular member; 320. a first annular member; 321. a communicating hole; 330. a second ring member.

Detailed Description

So that the manner in which the features and elements of the disclosed embodiments can be understood in detail, a more particular description of the disclosed embodiments, briefly summarized above, may be had by reference to the embodiments, some of which are illustrated in the appended drawings. In the following description of the technology, for purposes of explanation, numerous details are set forth in order to provide a thorough understanding of the disclosed embodiments. However, one or more embodiments may be practiced without these details. In other instances, well-known structures and devices may be shown in simplified form in order to simplify the drawing.

The terms "first," "second," and the like in the description and in the claims, and the above-described drawings of embodiments of the present disclosure, are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It should be understood that the data so used may be interchanged under appropriate circumstances such that embodiments of the present disclosure described herein may be made. Furthermore, the terms "comprising" and "having," as well as any variations thereof, are intended to cover non-exclusive inclusions.

In the embodiments of the present disclosure, the terms "upper", "lower", "inner", "middle", "outer", "front", "rear", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings. These terms are used primarily to better describe the disclosed embodiments and their examples and are not intended to limit the indicated devices, elements or components to a particular orientation or to be constructed and operated in a particular orientation. Moreover, some of the above terms may be used to indicate other meanings besides the orientation or positional relationship, for example, the term "on" may also be used to indicate some kind of attachment or connection relationship in some cases. The specific meanings of these terms in the embodiments of the present disclosure can be understood by those of ordinary skill in the art as appropriate.

In addition, the terms "disposed," "connected," and "secured" are to be construed broadly. For example, "connected" may be a fixed connection, a detachable connection, or a unitary construction; can be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements or components. Specific meanings of the above terms in the embodiments of the present disclosure can be understood by those of ordinary skill in the art according to specific situations.

The term "plurality" means two or more unless otherwise specified.

It should be noted that, in the case of no conflict, the embodiments and features in the embodiments of the present disclosure may be combined with each other.

As shown in connection with FIGS. 1-17, embodiments of the present disclosure provide a removable/assemblable co-culture organ chip, comprising a chip platform 100 and a culture chamber module 200. One or more central liquid storage holes 10 are formed in the chip platform 100, and the central liquid storage holes 10 are in a step shape; and the bottom wall of the central liquid storage hole 10 is provided with a mounting position; and a co-culture channel 30 is axially extended on the stepped surface 11 of the central liquid storage hole 10. The culture chamber module 200 is detachably arranged at the installation position of the bottom wall of the central liquid storage hole 10; one or more culture compartments are provided on the culture chamber module 200.

The assembled co-culture organ chip of the disclosed embodiment is based on the integrally designed co-culture organ chip, and the region on the bottom wall of the central reservoir well 10, which includes the culture partition (e.g., culture well 20 shown in FIG. 1), is divided to form a culture chamber module 200 (shown in FIG. 2), so that the region can be flexibly replaced. Therefore, the culture chamber modules 200 with the appropriate shape, number and size of the culture partitions can be selected and assembled on the chip platform 100 according to actual requirements, so that the requirement of accurate experimental design can be met, flexibility and variability are achieved, application scenes of the organ chips are increased, and the application upper limit of the organ chips is improved.

The assembled co-culture organ chip of the embodiment of the disclosure is open, and the openings of the culture partition and the co-culture channel 30 face to the same side, so that cell planting can be completed on the same side (e.g., upper side) of the organ chip, thereby greatly reducing the operation difficulty. By carrying out standardized design on the arrangement layout of the central liquid storage holes 10 on the chip platform 100 of the co-culture organ chip, the open type operation channel has strong compatibility with the commercial operation and detection equipment, is convenient for subsequent on-board detection and cell recovery to carry out analysis such as RNA (ribonucleic acid), protein extraction and the like, is simpler and more suitable for industrial popularization. The method is simple to operate, does not need professional technicians, enlarges the application range of the organ chip and improves the universality.

In the disclosed embodiment, a central reservoir 10 and a culture chamber module 200 detachably disposed therein constitute a culture unit. One or more culture units are provided on the chip platform 100.

In the embodiment of the present disclosure, the central liquid storage hole 10 may adopt a through hole structure, and a port at the bottom end of the central liquid storage hole 10 is an installation position. The culture cell module 200 is detachably disposed on the port of the bottom end of the central liquid storage hole 10. It is understood that when the central reservoir 10 is a blind hole, the bottom wall of the central reservoir 10 is divided, and one or more culture partitions (e.g., culture wells 20) are disposed on the bottom wall to form the culture chamber module 200, so that the bottom wall can be flexibly replaced.

In the embodiment of the present disclosure, the culture chamber module 200 may be detachably assembled to the chip platform 100 by using a colloid bonding method, a mortise-tenon structure embedding method, or a chemical bonding method.

Optionally, the tenon-and-mortise structures are adopted for embedding. Specifically, a protruding tenon (mortise) is provided on the peripheral wall of the culture chamber module 200, and a recessed mortise (mortise) is provided on the inner peripheral wall of the mounting position (or bottom end port) of the central liquid storage hole, and the tenon and the mortise are engaged, so that the culture chamber module 200 is detachably disposed on the mounting position (or bottom end port).

Alternatively, a chemical bonding method is used, specifically, the culture chamber module 200 is made of Polydimethylsiloxane (PDMS) material, and after the oxygen plasma treatment, the chemical bonding can be performed with the smooth surface of the chip platform made of glass or self material, so as to form a stable co-culture chamber structure.

In the embodiment of the present disclosure, the central liquid storage hole 10 of the culture unit is stepped, and includes a large aperture section 12 and a small aperture section 13, which are communicated with each other, and the large aperture section 12 is located on the upper layer. Then, when the large pore diameter section 12 of the central reservoir 10 is filled with culture medium or drug diluent, it can provide the required nutrient solution or drug to be tested for the cells/organs within the culture well 20 and co-culture channel 30; when only the small-aperture section 12 of the central reservoir 10 is filled with the culture medium or the drug diluent, it only provides the required nutrient solution or the drug to be tested for the cells/organs in the culture well 20, i.e. the small-aperture section 12 of the central reservoir 10 is the exclusive reservoir of the culture well 20. When the liquid level of the culture medium or the drug diluent filled in the co-culture channel 30 does not exceed the stepped surface 11 of the central liquid storage hole 10, the upper layer of the co-culture channel 30 is the exclusive liquid storage hole of the lower culture section.

In the detachable/assemblable co-culture organ chip of the embodiment of the present disclosure, the number, specification, and the like of the culture chamber modules 200 are not limited, and suitable culture chamber modules 200 are assembled to the chip platform 100 as required. Alternatively, the plurality of culture chamber modules 200 may include at least the culture chamber modules 200 corresponding to the number of the central wells 10 on the chip platform 100, but of course, the culture compartments of each culture chamber module 200 may be the same or different in number, shape, or combination thereof in the plurality of culture chamber modules 200.

In the embodiment of the present disclosure, cells/organs planted in different culture partitions in the central liquid storage hole 10 can be cultured independently, and different cells/organs planted in the culture partitions (e.g., culture holes 20) and the co-culture channel 30 can be co-cultured in a non-contact manner, so that the detachable/assemblable co-culture organ chip can realize the co-culture in a non-contact manner of cells or tissues, can be used for in vitro construction and long-term culture of multi-cell and multi-organ models, and further can be used for in vitro construction of multi-cell co-culture models or multi-organ co-culture models.

In the embodiment of the present disclosure, one or more culture chamber modules 200 of one or more specifications are respectively assembled on a plurality of central liquid storage wells 10 of one chip platform 100. The specifications of the culture chamber module 200 include one or more of the number of culture sections, the size of the culture sections, the shape of the culture sections, the size of the culture chamber module 200, and the shape of the culture chamber module 200, among others. By assembling culture chamber modules 200 of various specifications on one chip platform 100, tests for verifying the differences of different culture partitions cultured under the same culture environment conditions on samples can be performed.

In the embodiment of the present disclosure, the shape of the culture section is not limited, and may be one or more culture holes 20 provided in the culture chamber module 200 (as shown in FIGS. 1 and 3), or one or more culture regions 210 defined by the partition ribs 211 in the culture chamber module 200 (as shown in FIGS. 4 and 5).

The shape of the culture well 20 is not limited, and may be a circular hole, an oval, a square, a rectangle, a sector, or a polygon (e.g., a hexagon, an octagon, etc.), and the shape of the culture well 20 is designed so as to facilitate growth when the design requirement is satisfied. The number of the culture wells 20 is not limited, and may be 1, 2, 3, 4, 5 or more, and the specific number may be determined according to actual needs. The plurality of culture wells 20 may be arranged in an array, for example, a circular array or a rectangular array. As shown in FIG. 1, the culture well 20 is in the shape of a circular hole.

Alternatively, as shown in FIGS. 4 and 5, the separation ribs 211 are arranged in such a way that they cross the center of the culture chamber module 200, defining one or more culture areas 210 on the surface of the culture chamber module 200 or in the through holes.

Multiple culture compartments (e.g., culture wells 20 and culture regions 210) can be seeded with the same or different types of cells, and if the same cells are seeded, selective seeding can be performed depending on the amount of sample, for example, seeding only one of the regions (e.g., one culture well 20) can suffice. The co-culture channel 30 is used for culturing the cells of the second organ, the upper layer of the co-culture channel is used as a dedicated stock solution channel, and the bottom part/bottom wall of the co-culture channel is used for planting and culturing the cells of the second organ. The co-culture channel 30 may also be seeded with cells of the same or different types, depending on the actual requirements.

In the embodiment of the present disclosure, the culture section on the culture chamber module 200 is a through-hole (the culture well 20 shown in FIG. 2 is a through-hole) or a blind-hole. Wherein, when the culture partition is a through-hole, the co-culture organ chip further comprises a bottom plate disposed at the bottom of the culture well 20 and the co-culture channel 30. When in use, the utility model is placed on the bottom plate.

In the disclosed embodiment, the co-culture channel 30 is a through hole or a blind hole. Wherein, when the culture well 20 is a through-hole, the co-culture organ chip further comprises a bottom plate disposed at the bottom of the culture well 20 and the co-culture channel 30. When in use, the utility model is placed on the bottom plate.

In the disclosed embodiment, co-cultivation channel 30 comprises a closed/non-closed annular slot along the circumference of stepped surface 11 of central liquid storage well 10. I.e., co-cultivation channel 30 is a connected integral structure. In this embodiment, when the co-cultivation channel 30 comprises a closed circular slot, it is a blind hole. The structural integrity is ensured.

Alternatively, as shown in FIG. 1, a non-closed annular slot is axially extended from the stepped surface 11 of the central liquid storage hole 10 to serve as a co-culture channel 30.

Optionally, a closed annular slot is axially extended from the stepped surface 11 of the central liquid storage hole 10 to serve as a co-culture channel 30. In this embodiment, the annular slot may be a circular slot, which is a blind hole. The integrity of the organ chip body is ensured.

In the disclosed embodiment, the co-culture channel 30 is not limited to the above structure, and may include a plurality of pores. In this embodiment, the plurality of channels are circumferentially distributed along the stepped surface 11 of the central liquid storage hole 10. The cross section of the pore channel is not limited, and may be circular (as shown in fig. 6), oval, square, rectangular, fan-shaped (as shown in fig. 7), or polygonal (such as hexagon, octagon, etc.), and the shape of the pore channel is designed to contain as much culture medium or drug diluent as possible under the condition of meeting the design requirement.

In the disclosed embodiment, the co-culture channel 30 includes an upper liquid storage section 301 and a lower culture section 302 which are communicated; the size of the lower culture section 302 is smaller than or equal to the size of the upper reservoir section 301. When equal, the co-culture channel 30 is a straight channel (see FIGS. 2 and 9), and is easy to form. When the ratio is smaller than the above, the co-culture channel 30 is a step-shaped channel, i.e., the cross section thereof is step-shaped (as shown in fig. 8 and 10), so that the loss of the inoculated cells can be avoided, and the inoculation rate can be increased.

In some embodiments, as shown in fig. 8-13, in the chip platform 100, the central reservoir well 10 includes a central reservoir straight well 150 and a culture channel module. The culture channel module is detachably arranged in the central liquid storage straight hole 150, and the first end of the culture channel module is flush with the bottom end of the central liquid storage straight hole 150 and is provided with an installation position; wherein, the end face (equivalent to the stepped surface 11) of the second end of the culture channel module is provided with a co-culture channel 30 extending along the axial direction, or the culture channel module is matched with the central liquid storage straight hole 150 to form the co-culture channel 30. In this embodiment, the step portion of the step-shaped central liquid storage hole 10, in which the co-culture channel 30 is disposed, is divided, so that the step portion forms an independent module, which can be detached and assembled. That is, the co-culture channel 30 is also designed to be detachable to form an independent culture channel module, which can be flexibly replaced, and when in use, the culture channel module with the co-culture channel 30 of the proper specification is selected to be assembled into the central liquid storage straight hole 150. In this embodiment, the forming process of the chip platform 100 is simplified, the chip platform 100 may be a plate body, and one or more straight holes are formed thereon as the central liquid storage straight hole 150, so that the structure is simple and the forming is easy. When in use, the culture channel module and the culture chamber module 200 are assembled into the straight hole.

Optionally, the culture channel module is detachably disposed in the central liquid storage straight hole 150 by using a colloid bonding, mortise and tenon structure embedding or chemical bonding.

Alternatively, mortise and tenon structure engagement (not shown) is adopted. Set up convex tenon (tenon) on the perisporium of cultivateing the passageway module, set up concave mortise (fourth of the twelve earthly branches) on the internal perisporium of central stock solution straight hole, tenon and fourth of the twelve earthly branches interlock to will cultivate passageway module detachably and set up in central stock solution straight hole.

Optionally, a chemical bonding manner is adopted, specifically, the culture channel module is made of Polydimethylsiloxane (PDMS) material, and after oxygen plasma treatment, chemical bonding can be performed with the smooth surface of the chip platform made of glass or self material, so as to form a stable co-culture unit structure.

In some embodiments, as shown in fig. 8, the first culture channel module 300 has a cylindrical shape, and is provided with a through hole (i.e. the small-aperture section 13 of the central liquid storage hole 10) along the axial direction, the port of the first end is used as a mounting position, and the end wall of the second end is provided with a co-culture channel 30 extending along the axial direction. In this embodiment, the culture channel module is disassembled/assembled as a separate module, and the culture chamber module 200 is disassembled/assembled on the first end of the culture channel module 300. The outer side wall of the first culture channel module is in matched contact with the inner wall of the central liquid storage straight hole 150, and can be connected through colloid bonding or tenon-and-mortise embedding and the like.

In some embodiments, the second culture channel module comprises a ring member 310, wherein the ring member 310 is detachably disposed in the central liquid storage hole 150 in a manner of contacting with the inner wall of the central liquid storage hole 150 or in a manner of a set interval, and cooperates with the central liquid storage hole 10 (e.g., the inner wall thereof) to form the co-culture channel 30; and has a mounting location disposed on a first end thereof (i.e., the end that is flush with the bottom end of the central liquid-storing straight bore 150). In this embodiment, the outer sidewall of the annular member 310 (opposite the inner wall of the central reservoir 150) may be shaped according to the configuration and shape of the co-culture channel 30.

In this embodiment, when the annular member 310 and the inner wall of the central liquid-storing straight hole 150 are disposed at a predetermined interval, the shape of the annular member 310 and the shape of the central liquid-storing straight hole 150 may be the same or different, and is not limited.

Alternatively, as shown in fig. 9, the first annular member 310 may be detachably disposed in the central liquid storage hole 150 such that an outer sidewall thereof is linear and spaced apart from an inner wall of the central liquid storage hole 150 by a predetermined distance. The co-culture channel 30 constructed in this embodiment is a straight slot, i.e., the lower culture section 302 has a size equal to the size of the upper reservoir section 301. In addition, the co-culture channel 30 constructed in this embodiment is a non-closed circular slot, and the non-closed position may be a detachable connection position of the circular member 310 and the central straight liquid storage hole 150. In this embodiment, the setting interval is the radial width of the co-culture channel 30, and is determined according to actual needs.

Optionally, a second ring member 310, the outer side wall being stepped; the outer wall of the bottom step is in contact with the inner wall of the central liquid storage straight hole 150 or is detachably arranged in the central liquid storage straight hole 150 at a set interval. In the second ring member 310 of this embodiment, the bottom step is the largest step in the size of the ring member.

In this embodiment, when the outer wall of the bottom step of the second annular member 310 contacts the inner wall of the central liquid-storing straight hole 150 and is detachably disposed in the central liquid-storing straight hole 150, the constructed co-culture channel 30 is a closed annular slot and is a blind hole. Depending on the number of steps in the stepped outer side wall, the co-cultivation channel 30 is made to be a straight channel or a stepped channel.

In this embodiment, when the outer wall of the bottom step of the second annular component 310 and the inner wall of the central liquid storage straight hole 150 are detachably disposed in the central liquid storage straight hole 150 at a predetermined interval, the constructed co-culture channel 30 is a step-shaped channel, that is, the size of the lower culture section 302 of the co-culture channel 30 is smaller than that of the upper liquid storage section 301, so as to avoid the loss of the inoculated cells and improve the inoculation rate. In this embodiment, the spacing is set to the radial width of the lower culture section 302 of the co-culture channel 30, which is determined according to actual needs.

In some embodiments, a third culture channel module comprises a first ring member 320 and a second ring member 330. The first ring member 320 is detachably disposed in the central liquid-storing straight hole 150 and the outer wall thereof is in contact with the inner wall of the central liquid-storing straight hole 150. The second annular member 330 is disposed inside the first annular member 320, and is connected to the first annular member 320 to form the co-culture channel 30. A mounting location is provided at a first end thereof (the end flush with the bottom end of the central liquid storage straight bore 150). Wherein, the first ring member 320 is fixedly connected or detachably connected with the second ring member 330. In this embodiment, when the second annular member 330 is fixedly connected to the first annular member 320, the structure can be the same as the first culture channel module (shown in FIG. 8), which is an integral and independent module; when the two are detachably connected, the culture channel module is divided, so that the structure of the co-culture organ chip is more flexible, and more application scenes are provided.

In this embodiment, the second ring 330 may have the same structure as the ring 310, and is defined by a first second ring 330 (with a linear outer sidewall) and a second ring 330 (with a stepped outer sidewall). And will not be described in detail herein.

Optionally, the second ring member 330 is detachably sleeved inside the first ring member 320 in a manner of contacting with an inner wall of the first ring member 320 or forming a set interval. The outer sidewall of the second ring 330 opposes the inner sidewall of the first ring 320 to create the co-culture channel 30.

Alternatively, the inner side wall of the first ring member 320 may be linear or stepped. The first ring member 320, the inner side wall of which is linear as shown in fig. 13, is referred to as a first ring member. The first ring member 320 having the stepped inner side wall as shown in fig. 10 to 12 is referred to as a second first ring member.

Optionally, a third culturing channel module comprises a first ring-shaped member 320 and a first second ring-shaped member 330, wherein the first second ring-shaped member 330 is detachably disposed inside the first ring-shaped member 320 at a predetermined interval from the inner wall of the first ring-shaped member 320. Reference may be made to the structure shown in fig. 2. The co-cultivation channel 30 is formed as a non-closed circular groove hole and is a through hole.

Optionally, another third culturing channel module comprises a first ring-shaped member 320 and a second ring-shaped member 330, wherein the second ring-shaped member 330 is in contact with the inner wall of the first ring-shaped member 320 or is detachably disposed inside the first ring-shaped member 320 at a predetermined interval. The co-cultivation channel 30 is formed as a non-closed circular groove hole and is a through hole.

Alternatively, another third culture channel module, as shown in fig. 10, includes a second first ring member 320 and a first second ring member 330, and the first second ring member 330 is detachably disposed inside the second first ring member 320 at a predetermined interval from the inner wall of the second first ring member 320. The cross-sectional shape of the constructed co-culture channel 30 is stepped and is a through-hole.

Alternatively, the first and second ring members 330 may be detachably disposed inside the second first ring member 320 in such a manner as to be in contact with the inner wall of the second first ring member 320. The co-culture channel 30 may be a straight slot or a stepped slot, and may be a blind hole.

Alternatively, another third culture channel module comprises a second first ring member 320 and a second ring member 330, wherein the first second ring member 330 is detachably disposed inside the first ring member 320 in a manner of contacting with the inner wall of the first ring member 320 (as shown in fig. 11) or at a set interval. In this embodiment, the height of the bottom step of the second first ring member 320 is the same as or different from the height of the bottom step of the second ring member 330; when different, co-culture channels 30 having stepped cross-sections and blind holes can be constructed.

In a detachable/assemblable co-culture organ chip according to an embodiment of the present disclosure, both the culture channel module and the culture chamber module 200 can be detached/assembled, and thus, after the size of the central liquid storage straight hole 150 of the chip platform 100 is determined, the size of the co-culture channel 30 (e.g., the radial width of the co-culture channel 30) can be adjusted by adjusting the size of the ring-shaped member 310 or the second ring-shaped member 330, so that the planting area ratio of the co-culture channel 30 and the culture hole 20 can be adjusted to indirectly adjust the ratio of the cells seeded in the culture channel module and in the culture chamber module 200, thereby being more flexible and variable. According to actual requirements, different culture channel modules and culture chamber modules 200 may be assembled on the chip platform 100.

In some embodiments, the culture chamber module 200 is fixedly attached to the culture channel module, forming a co-culture module; the co-culture module is removably disposed within the central reservoir bore 150. That is, the culture chamber module 200 is fixedly attached in the mounting position (or the first end port) of the first end of the culture channel module, or both are integrally formed. In this embodiment, the co-culture module forms an integral independent module, which is assembled into the central liquid storage straight hole 150 of the chip platform 100, thereby facilitating assembly and disassembly.

Alternatively, in the first co-culture module, the culture channel module comprises a ring 310, and the culture chamber module 200 is fixedly connected to the mounting position of the end of the ring 310 to constitute the co-culture module. In particular, the ring 310 is integrally formed with the culture chamber module 200, i.e. the co-culture module is in the form of an open petri dish structure with one or more culture compartments (e.g. culture wells 20) arranged in the bottom wall. The first co-cultivation module of this embodiment forms an integral, self-contained module. For example, culture chamber module 200 shown in FIG. 9 is fixedly attached to ring 310.

Alternatively, in the first co-culture module, as shown in FIG. 11, the culture channel module comprises a first ring member 320 and a second ring member 330, and the culture chamber module 200 is fixedly connected to the second ring member 330 to constitute a co-culture module. In particular, the second annular member 330 is integrally formed with the culture chamber module 200 to form an open culture dish structure. In this embodiment, the co-culture module forms an integral, self-contained module when the first ring member 320 and the second ring member 330 are fixedly attached, and the culture chamber module 200 is fixedly attached to the culture channel module 300 as shown with reference to FIG. 8. When the first ring member 320 and the second ring member 330 are removably coupled, the co-culture module includes the first ring member 320 and the culture dish structure that are removably coupled, as shown in FIGS. 10 and 11.

The area (or volume) of the co-culture channel 30, and thus the proportion of cells seeded in the culture well 20 of the culture chamber module 200 and the co-culture channel 30, can be adjusted by increasing or decreasing the size of the first co-culture module or the size of the culture dish structure (i.e. the area of the culture chamber module 200) in the second co-culture module.

In some embodiments, as shown in fig. 11, 12 and 13, when the culture channel module in the co-culture module comprises the first ring member 320, the second end of the first ring member 320 is extended, such that the first ring member 320 forms a liquid storage hole structure. The thickness of the chip platform 100 may be reduced. In this embodiment, the first ring member 320, the second ring member 330 and the culture chamber module 200 are extended to form a complete culture unit, so that the culture unit can be integrally and detachably assembled, and the culture unit can be used more flexibly.

In some embodiments, the culture channel module and culture chamber module 200 are fabricated from materials having one or more of biocompatibility, biomimetic, cell penetrability, and degradability. The module material with biocompatibility, bionic property or cell penetrability can better ensure the normal growth state of cells in an in vitro model and similar to the growth state in an organism while meeting the specific structural function, and reduces the influence of external factors on the bionic function of the in vitro model. The degradable module material improves the environmental friendliness of the module.

Alternatively, the culture channel module and the culture chamber module 200 are made of biocompatible materials by 3D printing technology.

Alternatively, the culture channel module and the culture chamber module 200 are prepared by laser engraving or the like.

In some embodiments, the culture cell module 200 is made from a hydrogel material. The application scene is expanded, and cell migration or transfer tests can be performed. Alternatively, the culture chamber module 200 is made from a soft hydrogel material. For example, methacrylated gelatin (gelma), alginate or collagen.

In some embodiments, as shown in FIG. 10, co-culture channel 30, comprises an upper reservoir section 301 and a lower culture section 302 in communication; can be detached from the side wall between the upper liquid storage section 301 and the central liquid storage hole 10 (or the central liquid storage straight hole 150). That is, the side wall of the co-culture channel 30 on the side of the culture chamber module 200 corresponding to the upper reservoir section 301 is detachable. This partial sidewall, like the enclosure 220 between the culture well 20 and the co-culture channel 30 in the culture chamber module 200, avoids mixed cross-contamination of the sample/culture fluid in the two module areas. This enclosure 220 is designed as detachable in this embodiment, can make the organ chip improve and cultivate to cultivate the component material exchange efficiency and avoid different regional sample mixture cross contamination's two kinds of advantages between nimble conversion. For example, when organoids are planted in the culture chamber module 200 and stromal cells are planted in the co-culture channel 30/static and dynamic co-culture, the enclosing wall is detached, and a mode without the enclosing wall is adopted, so that the elimination of metabolic wastes in the culture chamber can be better promoted, the influence of small molecules generated by stromal cells B in the co-culture channel 30 on cells A in the culture hole 20 of the culture chamber module 200 can be better promoted, the exchange efficiency of culture component substances can be better improved, and the advantages of a co-culture system can be achieved. When the downthehole cell A of culture is collected in the digestion, get rid of behind the organ chip culture medium, go the enclosure assembly, for example adopt the fixed enclosure of mortise and tenon joint structure gomphosis, isolated and the connection between the coculture passageway 30 in the outside avoids coculture passageway 30 in the matrix cell B to the cross mixed pollution who cultivates cell A in the cell, the guarantee to collect the purity degree of target cell.

Optionally, the ring member 310 or the second ring member 330 is divided into a detachable upper half and a detachable lower half.

In some embodiments, as shown in fig. 14 and 15, the chip platform 100 further includes a side liquid storage hole 40 and a communication channel 50, wherein a plurality of side liquid storage holes 40 are disposed around each central liquid storage hole 10; and the liquid storage hole at the side is a through hole or a blind hole. The communicating channel 50 communicates the central liquid storage hole 10 with the peripheral side liquid storage holes 40, and/or communicates the co-culture channel 30 with the peripheral side liquid storage holes 40 of the central liquid storage hole 10. The chip platform 100 of the present embodiment enables dynamic culture of co-culture organ chips, enables dynamic culture or dynamic co-culture of culture partitions (e.g., culture wells 20) and/or co-culture channels 30, and is compatible with a variety of fluid manipulation methods. Real-time dynamic updates of the culture environment within the culture zones and/or co-culture channels 30 may be achieved. In addition, cells may be planted in the side reservoir hole 40 to expand the planting area and increase the amount of cell culture.

The dynamic co-culture organ chip formed by the chip platform 100 of the embodiment has three communication modes, the first mode is that the first communication channel 51 is only communicated with the central liquid storage hole 10 and the peripheral side liquid storage holes 40; the second is that the second communicating channel 52 only communicates the coculture channel 30 and the paired side liquid storage holes 40 around the central liquid storage hole 10; the third is that the communicating channel 50 communicates with the central liquid storage hole 10 and the pair of side liquid storage holes 40 around the central liquid storage hole, and communicates with the coculture channel 30 and the pair of side liquid storage holes 40 around the central liquid storage hole 10. In the third communication mode, the communication passage 50 may be the first communication passage 51 and the second communication passage 52 which are independently provided, or may be one communication passage 50 in which the first communication passage 51 and the second communication passage 52 are integrated (as shown in fig. 14).

Accordingly, when the culture channel module is divided from the chip platform 100, communication holes are provided on the side walls of the culture channel module at the corresponding positions for communication with the communication channels 50. As shown in fig. 14, a communication hole 321 is provided in the first ring member 320.

In this embodiment, two or more side liquid storage holes 40 are formed around each central liquid storage hole 10, thereby realizing dynamic culture. Alternatively, the side reservoir holes 40 are provided in pairs around the central reservoir hole 10. The pair of side reservoirs 40 may be provided in plural, and is not limited to the pair of side reservoirs 40 shown in fig. 14. Improve the balance of fluid flow and improve the dynamic culture effect. A communication passage 50 (e.g., a first communication passage 51) is provided between the central reservoir hole 10 and each of the side reservoir holes 40 to communicate with each other. Furthermore, the number of the communication channels 50 between the central liquid storage hole 10 and each of the side liquid storage holes 40 is not limited to one as shown in FIG. 14, and a plurality of communication channels may be provided to improve the dynamic culture effect.

In this embodiment, the size of the communication channel 50 is designed to allow microfluidic control of the medium in the central reservoir 10 and/or the co-culture channel. The cross-sectional shape is also not limited and may be circular, square or other geometric shapes.

Optionally, as shown in fig. 15, the chip platform 100 further includes a side liquid storage hole 410 detachably disposed on the side liquid storage hole 40. In this embodiment, the volume of the side liquid storage hole 40 is increased, and the thickness of the chip platform 100 can be reduced.

Alternatively, the size of the side reservoir hole may be equal to or larger than the size of the side reservoir hole 40. When larger, more medium or drug diluent may be contained.

Alternatively, the side reservoir wells 40 are of the well size of a standard 48-well plate or a standard 96-well plate. The method is convenient for subsequent on-machine detection and cell recovery to perform analysis such as RNA and protein extraction, is simpler and more suitable for industrial popularization.

In the embodiment of the disclosure, a chip platform 100 may be an integrated structure. Optionally, the chip platform 100 is a plate body, and a plurality of central liquid storage holes 10 or central line liquid storage straight holes are formed on the plate body. For example, integral injection molding, or laser etching on the plate body, etc. is used. Specifically, Polystyrene (PS) and Polymethyl Methacrylate (PMMA) are integrally injection-molded, and the materials are low in cost, easy to injection mold, free of any toxicity to cells and free of specific adsorption.

In the disclosed embodiment, another chip platform 100 can be obtained by a layered processing and assembly configuration. The assembly is carried out after the layering processing, the whole part is broken into parts, the forming process is simplified, and the forming of the structure on the chip platform 100 is facilitated.

In some embodiments, the chip platform 100 including the side reservoirs 40 and the communication channels 50 is designed as a two-layer structure. As shown in fig. 16, the chip body includes an upper stage and a lower stage, the upper stage is provided with a plurality of first side liquid-storing holes 41 and first communicating channels 51, and the plurality of first side liquid-storing holes 41 are arranged in pairs around (e.g., on both sides of) the first liquid-storing holes; the first communication passage 51 communicates the first reservoir and the pair of first side reservoir 41 on both sides thereof, respectively; and/or, a plurality of second side liquid storage holes 42 and a plurality of second communication channels 52 are also arranged on the lower platform, and the plurality of second side liquid storage holes 42 are arranged around the second liquid storage holes in pairs (for example, on two sides); the second communicating passage 52 communicates the second reservoir holes and the pair of second side reservoir holes 42 on both sides thereof, respectively. Wherein, first stock solution hole and the coaxial setting in second stock solution hole form central stock solution hole 10 or central stock solution straight hole 150, and first limit side stock solution hole 41 and the coaxial setting in second limit side stock solution hole 42 form limit side stock solution hole 40.

The present embodiment corresponds to the aforementioned three communication manners, wherein for the third communication manner, the first communication passage 51 and the second communication passage 52 are independently disposed or can be buckled to form one communication passage 50 (as shown in fig. 16). The independent arrangement means that the first communicating channel 51 and the second communicating channel 52 are kept in an independent state and do not generate mixed flow even after the upper deck 110 and the lower deck 120 are fastened.

In the embodiment of the present disclosure, the two layers of the chip platform 100 may be bonded and assembled together by using a sealing process such as double-sided tape, ultrasonic bonding, thermal bonding, plasma, and thermal pressing.

In the embodiment of the present disclosure, as shown in fig. 17, a surrounding barrier 101 is disposed on a peripheral edge of the surface of the chip platform 100, that is, the surrounding barrier 101 is disposed on the peripheral edge of the surface of the chip platform 100. The enclosure 101 encloses to form a liquid containing tank 102, so that the evaporation of the culture medium in the culture process is reduced, and the culture effect is improved.

In the embodiment of the present disclosure, the chip platform and each culture module (e.g., the culture chamber module and the culture channel module) of the detachable/assemblable co-culture organ chip are made of PMMA, PS, etc. The culture chamber module can be prepared from hydrogel materials, so that the cell culture function of the culture chamber module is realized, and the culture chamber module can be conveniently disassembled/assembled. The chip structure of each layer can be obtained by soft lithography, molding method, laser etching, machining, LIGA or one-time injection molding.

In the embodiment of the present disclosure, the central liquid storage hole 10 or the central liquid storage straight hole 150 on the chip platform 100 may be a concave hole, as shown in fig. 8 to 10, and is disposed on the body of the chip platform 100. Alternatively, the pillar holes may be formed by, as shown in fig. 2, pillars protruding from the surface of the chip platform 100 and having hole structures formed thereon. Of course, the second end of the first ring 320 may extend upward to form a central liquid hole structure, so as to reduce the thickness of the chip platform 100.

In the embodiment of the present disclosure, the liquid storage hole on the side of the chip platform 100 may be a concave hole or a pillar hole. The method is not limited, and the method can be determined according to actual requirements.

The above description and drawings sufficiently illustrate embodiments of the disclosure to enable those skilled in the art to practice them. Other embodiments may include structural and other changes. The examples merely typify possible variations. Individual components and functions are optional unless explicitly required, and the sequence of operations may vary. Portions and features of some embodiments may be included in or substituted for those of others. The embodiments of the present disclosure are not limited to the structures that have been described above and shown in the drawings, and various modifications and changes may be made without departing from the scope thereof. The scope of the present disclosure is limited only by the appended claims.

Claims (10)

1. A removable/assemblable co-culture organ chip, comprising:

the chip platform is provided with one or more central liquid storage holes, the central liquid storage holes are in a step shape, the bottom walls of the central liquid storage holes are provided with mounting positions, and the step surfaces of the central liquid storage holes are axially extended and provided with co-culture channels;

the culture chamber module is detachably arranged at the mounting position of the bottom wall of the central liquid storage hole, and one or more culture partitions are arranged on the culture chamber module.

2. The detachable/assemblable co-culture organ chip of claim 1, wherein in the chip platform, the central reservoir well comprises:

a central liquid storage straight hole;

the culture channel module is detachably arranged in the central liquid storage straight hole, and the first end of the culture channel module is flush with the bottom end of the central liquid storage straight hole and is provided with an installation position;

the end face of the second end of the culture channel module is provided with the co-culture channel extending along the axial direction, or the culture channel module is matched with the central liquid storage straight hole to form the co-culture channel.

3. The detachable/assemblable co-culture organ chip according to claim 2,

the culture channel module comprises:

the annular piece is detachably arranged in the central liquid storage straight hole in a manner of contacting with the inner wall of the central liquid storage straight hole or presenting a set interval, and is matched with the central liquid storage straight hole to form a co-culture channel; and the first end of the annular member is provided with a mounting position;

alternatively, the culture channel module comprises:

the first annular piece is detachably arranged in the central liquid storage straight hole, and the outer wall of the first annular piece is in contact fit with the inner wall of the central liquid storage straight hole;

the second annular piece is sleeved in the first annular piece and is connected and matched with the first annular piece to form the co-culture channel; and the first end of the first connecting rod is provided with a mounting position;

wherein the first annular member is fixedly connected or detachably connected with the second annular member.

4. The detachable/assemblable co-culture organ chip according to claim 3,

the culture channel module comprises an annular piece, and the annular piece is annular and the outer side wall of the annular piece is linear or stepped;

or the culture channel module comprises a first annular piece, and the inner side wall of the first annular piece is linear or stepped;

or the culture channel module comprises a second ring-shaped member, and the outer side wall of the second ring-shaped member is linear or stepped.

5. The detachable/assemblable co-culture organ chip of claim 3, wherein the culture channel module comprises a first ring member, a second end of the first ring member being extended such that the first ring member forms a reservoir structure.

6. The detachable/assemblable co-culture organ chip of any one of claims 2 to 5, wherein the culture chamber module is fixedly connected to a mounting position of the culture channel module to constitute a co-culture module; the co-culture module is detachably arranged in the central liquid storage straight hole.

7. The detachable/assemblable co-culture organ chip according to any one of claims 1 to 5, wherein the culture chamber module is prepared using a hydrogel material;

alternatively, when the co-culture organ chip comprises a culture channel module, the culture channel module and the culture chamber module are prepared using a material having one or more of biocompatibility, biomimetic property, cell penetrability and degradability.

8. The detachable/assemblable co-culture organ chip according to any one of claims 1 to 5, wherein the co-culture channel comprises an upper reservoir section and a lower culture section in communication; the lateral wall between upper strata stock solution section and the central stock solution hole can be dismantled.

9. The detachable/assemblable co-culture organ chip of any one of claims 1 to 5, wherein the chip platform further comprises:

a plurality of lateral liquid storage holes arranged around the central liquid storage hole; the side liquid storage hole is a through hole or a blind hole;

and the communicating channel is communicated with the central liquid storage hole and the peripheral liquid storage holes thereof, and/or is communicated with the coculture channel and the peripheral liquid storage holes of the central liquid storage hole where the coculture channel is located.

10. The detachable/assemblable co-culture organ chip of claim 9, further comprising:

a plurality of avris stock solution post holes, detachably set up in on the avris stock solution hole.

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