Cell culture device

Abstract

This article discloses a cell culture device, which relates to the field of biotechnology, including: a culture dish, including a bottom plate, a surrounding plate and at least one partition, the surrounding plate is arranged along the circumference of the bottom plate, the bottom plate and the surrounding plate form a receiving groove, the partition is arranged in the receiving groove and defines at least one culture chamber, and there is a through hole between the partition and the bottom plate that is connected to the culture chamber; an elastic basement membrane is penetrated in the through hole and covers the position corresponding to the bottom plate and the culture chamber; an actuator is at least partially arranged in the receiving groove and located between the surrounding plate and the partition, and is used to stretch the elastic basement membrane. This article arranges an elastic basement membrane at the bottom of the culture chamber, and the elastic basement membrane can be stretched under the drive of the actuator. During the stretching process of the elastic basement membrane, the cells on the elastic basement membrane will also be stretched synchronously, thereby simulating the growth environment of cells in the body, and more realistically reflecting the state of cell culture in the body.

Description

Technical Field

[0001] The present application relates to, but is not limited to, the field of biotechnology, and more specifically, to a cell culture device. Background Art

[0002] Cell culture is the basic unit operation in biological research. Microfluidic chips rely on microfabrication technology to construct two-dimensional or three-dimensional network structures at the micrometer or even nanometer level, which can integrate functional units such as micropumps, microvalves and microreactors. Microfluidic cell culture has the characteristics of reducing sample volume and being able to customize and study multiple microenvironments in the same culture device.

[0003] Under physiological conditions, cells are in a dynamic equilibrium environment. For example, vascular endothelial cells under physiological conditions are usually subjected to three mechanical forces: hydrostatic pressure, shear force generated by blood flow, and tensile force generated during vasodilation and contraction. However, in traditional in vitro cell culture, cells are in a relatively static environment, which is very different from the in vivo environment. In vitro cell culture cannot truly reflect the state of cells in vivo. Summary of the invention

[0004] The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.

[0005] The embodiment of the present invention provides a cell culture device, which can simulate the growth environment of cells in vivo and more realistically reflect the state of cell culture in vivo.

[0006] The cell culture device provided in the embodiment of the present invention comprises:

[0007] A culture dish, comprising a bottom plate, a surrounding plate and at least one partition plate, wherein the surrounding plate is arranged along the circumference of the bottom plate, the bottom plate and the surrounding plate enclose a receiving groove, the partition plate is arranged in the receiving groove and defines at least one culture chamber, and a through hole communicating with the culture chamber is provided between the partition plate and the bottom plate;

[0008] The elastic basement membrane is penetrated in the penetration hole and covers the corresponding position of the bottom plate and the culture chamber;

[0009] The actuator is at least partially disposed in the accommodating groove and located between the enclosure plate and the partition plate, and is used for stretching the elastic base membrane.

[0010] In some exemplary embodiments, the partition includes a first partition, the first partition includes a first sub-partition extending along a first direction, and a second sub-partition and a third sub-partition arranged on the first sub-partition along the first direction and extending along a second direction, the first sub-partition is arranged on a bottom plate, there is a gap between the second sub-partition and the bottom plate, there is a gap between the third sub-partition and the bottom plate, the through holes include a first through hole between the second sub-partition and the bottom plate, and a second through hole between the third sub-partition and the bottom plate, and the first direction and the second direction intersect.

[0011] In some exemplary embodiments, in a direction perpendicular to the bottom plate, the diameter of the through hole is adapted to the thickness of the elastic base membrane, and in a second direction, the diameter of the through hole is adapted to the width of the elastic base membrane.

[0012] In some exemplary embodiments, the first partition includes a plurality of first partitions spaced apart along the second direction.

[0013] In some exemplary embodiments, the enclosure includes a first sub-enclosure and a second sub-enclosure extending in a first direction and spaced apart in a second direction, and a third sub-enclosure and a fourth sub-enclosure extending in the second direction and spaced apart in the first direction, the third sub-enclosure is adjacent to the second sub-partition, the fourth sub-enclosure is adjacent to the third sub-partition, a first mounting groove connected to the first through hole is formed between the third sub-enclosure and the second sub-partition, and a second mounting groove connected to the second through hole is formed between the fourth sub-enclosure and the third sub-partition.

[0014] In some exemplary embodiments, the actuator includes a first roller installed in a first mounting groove and a second roller installed in a second mounting groove, and the elastic base membrane includes a first end and a second end spaced apart along a first direction, the first end is connected to the first roller, and the second end is connected to the second roller.

[0015] In some exemplary embodiments, the first roller includes a first shaft body and a first clamping shaft arranged at both ends of the first shaft body, two first clamping seats are arranged in the first installation groove, the first shaft body is located between the two first clamping seats, and the first clamping shaft is seated in the corresponding first clamping seat, and the second roller includes a second shaft body and a second clamping shaft arranged at both ends of the second shaft body, two second clamping seats are arranged in the second installation groove, the second shaft body is located between the two second clamping seats, and the second clamping shaft is seated in the corresponding second clamping seat.

[0016] In some exemplary embodiments, the first sub-partition of the first partition adjacent to the first sub-enclosure extends into the first mounting groove to form a first extension portion, and extends into the second mounting groove to form a second extension portion, one of the two first sockets includes a first extension portion and a first slot disposed on an end surface of the first extension portion away from the bottom plate, one of the two second sockets includes a second extension portion and a second slot disposed on an end surface of the second extension portion away from the bottom plate, the first card shaft is embedded in the first card slot and can rotate, and the second card shaft is embedded in the second card slot and can rotate.

[0017] In some exemplary embodiments, the partition also includes a second partition arranged between the first partition and the second sub-enclosure, the second partition includes a third extension portion extending into the first mounting groove and a fourth extension portion extending into the second mounting groove, the other of the two first sockets includes a third extension portion and a third socket arranged on the end surface of the third extension portion away from the bottom plate, the other of the two second sockets includes a fourth extension portion and a fourth socket arranged on the end surface of the fourth extension portion away from the bottom plate, the first socket is embedded in the third socket and can rotate, and the second socket is embedded in the fourth socket and can rotate.

[0018] In some exemplary embodiments, a first axial hole connected to the first mounting groove and a second axial hole connected to the second mounting groove are provided on a surface of one of the first sub-enclosure and the second sub-enclosure away from the accommodating groove, and the actuator includes a first drive shaft passed through the first axial hole and a second drive shaft passed through the second axial hole, a first mating structure is provided at one end of the first drive shaft adjacent to the first clamping shaft, a second mating structure engaged with the first mating structure is provided on the first clamping shaft adjacent to the first drive shaft, the first drive shaft is configured to be able to actuate the first roller after the first mating structure and the second mating structure are engaged, a third mating structure is provided at one end of the second drive shaft adjacent to the second clamping shaft, a fourth mating structure engaged with the third mating structure is provided on the second clamping shaft adjacent to the second drive shaft, and the second drive shaft is configured to actuate the second roller after the third mating structure and the fourth mating structure are engaged.

[0019] In some exemplary embodiments, a first actuator is provided at one end of the first drive shaft away from the first roller, and a second actuator is provided at one end of the second drive shaft away from the second roller. The first actuator is configured to drive the first drive shaft to actuate the first roller to rotate, and the second actuator is configured to drive the second drive shaft to actuate the second roller to rotate.

[0020] In some exemplary embodiments, a fifth mating structure corresponding to the position of the first installation groove and a sixth mating structure corresponding to the position of the second installation groove are provided on the surface of the other of the first sub-enclosure and the second sub-enclosure facing the accommodating groove, a seventh mating structure rotatably connected to the fifth mating structure is provided on the first clamping shaft adjacent to the fifth mating structure, and an eighth mating structure rotatably connected to the sixth mating structure is provided on the second clamping shaft adjacent to the sixth mating structure.

[0021] In some exemplary embodiments, a fifth slot is provided on the first roller, a sixth slot is provided on the second roller, a first clip cooperating with the fifth slot is provided at the first end of the elastic base membrane, and a second clip cooperating with the sixth slot is provided at the second end of the elastic base membrane.

[0022] In some exemplary embodiments, a first limiting groove is provided at the slot position of the fifth slot, and a second limiting groove is provided at the slot position of the sixth slot. In the second direction, the width of the fifth slot is greater than the width of the first limiting groove, the width of the first limiting groove is adapted to the width of the elastic base membrane, and is configured to limit the movement of the elastic base membrane in the second direction. The width of the sixth slot is greater than the width of the second limiting groove, the width of the second limiting groove is adapted to the width of the elastic base membrane, and is configured to limit the movement of the elastic base membrane in the second direction.

[0023] In some exemplary embodiments, the cell culture device also includes a cover plate, which is arranged on the outer side of the surrounding plate and covers the containing tank. The cover plate includes a top plate and a side plate arranged on the circumference of the top plate. The side plate is located on the side of the surrounding plate away from the containing tank. A liquid exchange port is arranged on the top plate, and the liquid exchange port corresponds to the culture chamber.

[0024] An embodiment of the present invention provides a cell culture device, which disposes an elastic basement membrane at the bottom of a culture chamber. The elastic basement membrane can be stretched under the drive of an actuator. During the stretching of the elastic basement membrane, the cells on the elastic basement membrane will also be stretched, thereby simulating the growth environment of cells in the body and more realistically reflecting the state of cell culture in the body.

[0025] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention can be realized and obtained by the structures particularly pointed out in the description, claims and drawings.

[0026] Other aspects will be apparent upon reading and understanding the drawings and detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The accompanying drawings are used to provide a further understanding of the technical solution of the present invention and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the technical solution of the present invention and do not constitute a limitation on the technical solution of the present invention.

[0028] Figure 1 is a perspective view of a cell culture device according to an exemplary embodiment of the present invention;

[0029] Figure 2 An exploded view of a cell culture device according to an exemplary embodiment of the present invention;

[0030] Figure 3a A structural diagram of a culture dish according to an exemplary embodiment of the present invention;

[0031] Figure 3b for Figure 3a Cross-section view at the middle AA position;

[0032] Figure 4 is a plan view of a first card slot of an exemplary embodiment of the present invention;

[0033] Figure 5 is a plan view of a second card slot of an exemplary embodiment of the present invention;

[0034] Figure 6 It is a structural diagram of the third card slot position of an exemplary embodiment of the present invention;

[0035] Figure 7 is a partial enlarged view of a first roller of an exemplary embodiment of the present invention;

[0036] Figure 8 for Figure 7 Cross-section view at the middle AA position;

[0037] Figure 9a A schematic diagram of an elastic basement membrane of a cell culture device according to an exemplary embodiment of the present invention in an unstretched state;

[0038] Figure 9b FIG. 1 is a schematic diagram of an elastic basement membrane of a cell culture device according to an exemplary embodiment of the present invention in a stretched state. DETAILED DESCRIPTION

[0039] In order to make the purpose, technical solution and advantages of the present invention more clear, the embodiments of the present invention will be described in detail with reference to the accompanying drawings. It should be noted that the embodiments and features in the embodiments of the present application can be combined with each other arbitrarily without conflict.

[0040] In the description of the present invention, it should be noted that the terms "upper", "lower", "one side", "the other side", "one end", "the other end", "side", "relative", "four corners", "periphery", "'mouth'-shaped structure" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred structure has a specific orientation, is constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0041] In the description of the embodiments of the present invention, unless otherwise clearly specified and limited, the terms "connection", "direct connection", "indirect connection", "fixed connection", "installation", and "assembly" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; the terms "installation", "connection", and "fixed connection" can be a direct connection, or an indirect connection through an intermediate medium, or the internal connection of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0042] The cell culture device provided in the embodiment of the present invention comprises:

[0043] A culture dish, comprising a bottom plate, a surrounding plate and at least one partition plate, wherein the surrounding plate is arranged along the circumference of the bottom plate, the bottom plate and the surrounding plate enclose a receiving groove, the partition plate is arranged in the receiving groove and defines at least one culture chamber, and a through hole communicating with the culture chamber is provided between the partition plate and the bottom plate;

[0044] The elastic basement membrane is penetrated in the penetration hole and covers the corresponding position of the bottom plate and the culture chamber;

[0045] The actuator is at least partially disposed in the accommodating groove and located between the enclosure plate and the partition plate, and is used for stretching the elastic base membrane.

[0046] An embodiment of the present invention provides a cell culture device, which arranges an elastic basement membrane at the bottom of a culture chamber. The elastic basement membrane can be stretched under the drive of an actuator. During the stretching of the elastic basement membrane, the cells attached to the elastic basement membrane will also be stretched synchronously, thereby simulating the growth environment of cells in the body and more realistically reflecting the state of cell culture in the body.

[0047] The technical solutions of the exemplary embodiments of the present invention are exemplarily described below with reference to the accompanying drawings.

[0048] Figure 1 is a perspective view of a cell culture device according to an exemplary embodiment of the present invention, Figure 2: is an exploded view of a cell culture device of an exemplary embodiment of the present invention. In some exemplary embodiments, the cell culture device 1 is used for in vitro cell culture, and provides a tensile force to the cells during the cell culture process to simulate the growth environment of the cells in vivo. Therefore, the cell culture device 1 mainly includes a culture dish 100, an elastic basement membrane 200 and an actuator 300. The culture dish 100 is used to provide a place and environment for in vitro cell culture. The culture dish 100 includes a bottom plate 110, a surrounding plate 120 and at least one partition 130, the surrounding plate 120 is arranged along the circumference of the bottom plate 110, the bottom plate 110 and the surrounding plate 120 form a receiving groove 140, the partition 130 is arranged in the receiving groove 140 and defines at least one culture chamber 150, and there is a through hole between the partition 130 and the bottom plate 110 that is connected to the culture chamber 150, and the culture dish 100 can be made of a resin material, such as polypropylene (PP), polyethylene (PE) or polymethyl methacrylate (PET). The elastic basement membrane 200 has certain tensile properties, and cells can be attached to the elastic basement membrane 200. The elastic basement membrane 200 can be made of polydimethylsiloxane (PDMS), which has biocompatibility and good elasticity, and can be used for cells to attach and grow thereon. The elastic basement membrane 200 is penetrated in the penetration hole and covers the corresponding position of the bottom plate 110 and the culture chamber 150. The actuator 300 is at least partially disposed in the receiving groove 140 and is located between the enclosure 120 and the partition 130. The actuator 300 is connected to the end of the elastic basement membrane 200 and is used to stretch the elastic basement membrane 200. When the actuator 300 stretches the elastic basement membrane 200, the cells attached to the elastic basement membrane 200 will be stretched synchronously, thereby simulating the effect of the tensile force generated during the vasoconstriction and vasodilation of the cells in the body. According to Hooke's law, when a solid material is subjected to force, the stress and strain of the material are in a linear relationship, that is, F = -k·x, where k is the elastic coefficient, which is only related to the material properties, x is the deformation length, and F is the elastic force. Therefore, when the actuator 300 stretches the elastic basement membrane 200 to a specific length, a specific elastic force can be generated, and the cells are subjected to a specific tensile force on the elastic basement membrane 200.

[0049] The present invention exemplarily provides a cell culture device 1, which disposes an elastic base membrane 200 at the bottom of a culture chamber 150. The elastic base membrane 200 can be stretched under the drive of an actuator 300. During the stretching of the elastic base membrane 200, cells attached to the elastic base membrane 200 will also be stretched synchronously, thereby simulating the growth environment of cells in the body and more realistically reflecting the state of cell culture in the body.

[0050] Figure 3a is a structural diagram of a culture dish 100 according to an exemplary embodiment of the present invention. Figure 3b for Figure 3a In some exemplary embodiments, as shown in FIG. Figure 3aand 3b As shown, the partition 130 includes a first partition 131, the first partition 131 includes a first sub-partition 132 extending along a first direction and a second sub-partition 133 and a third sub-partition 134 spaced apart along the first direction on the first sub-partition 132 and extending along a second direction, the first sub-partition 132 is arranged on the bottom plate 110, there is a gap between the second sub-partition 133 and the bottom plate 110, there is a gap between the third sub-partition 134 and the bottom plate 110, the through hole 160 includes a first through hole 161 between the second sub-partition 133 and the bottom plate 110 and a second through hole between the third sub-partition 134 and the bottom plate 110 (not shown in the drawings), the first direction intersects with the second direction, and the first direction is as shown in FIG. Figure 3a The X direction is shown, and the second direction is as Figure 3a In the Y direction shown, the intersection of the first direction and the second direction includes the first direction being perpendicular to the second direction, where the perpendicularity can be substantially perpendicular, and the angle between the first direction and the second direction can be 87°, 90°, or 93°. In the direction perpendicular to the bottom plate 110, the aperture of the perforated hole 160 is adapted to the thickness of the elastic base film 200, and in the second direction, the aperture of the perforated hole 160 is adapted to the width of the elastic base film 200. The adaptation here can be understood as: in the direction perpendicular to the bottom plate 110, the aperture of the perforated hole 160 is equal to or slightly larger than the thickness of the elastic base film 200, where the aperture of the perforated hole 160 is 1.01 to 1.2 times the thickness of the elastic base film 200. In the second direction, the aperture of the perforated hole 160 is equal to or slightly larger than the width of the elastic base film 200, where the aperture of the perforated hole 160 is 1.01 to 1.2 times the width of the elastic base film 200. The aforementioned penetration hole 160 may refer to a first penetration hole 161 and a second penetration hole.

[0051] In some exemplary embodiments, Figure 3a and 3bAs shown, the first partition 131 includes a plurality of partitions spaced apart along the second direction. The number of the elastic base membranes 200 can be the same as the number of the first partitions 131, and the positions correspond one to one. Two first partitions 131 adjacent to each other along the second direction can be defined as a first front partition 131a and a first rear partition 131b, wherein the second sub-partition 133 and the third sub-partition 134 of the first front partition 131a are adjacent to the first sub-partition 132 of the first rear partition 131b, wherein a first penetration channel 162 communicating with the first penetration hole 161 is formed between the second sub-partition 133 of the first front partition 131a and the first sub-partition 132 of the first rear partition 131b, and a second penetration channel 163 communicating with the second penetration hole (not shown in the drawings) is formed between the third sub-partition 134 of the first front partition 131a and the first sub-partition 132 of the first rear partition 131b. The elastic base membrane 200 may be penetrated to the first penetration hole 161 through the first penetration channel 162 , and penetrated to the second penetration hole through the second penetration channel 163 .

[0052] In some exemplary embodiments, Figure 3a As shown, the enclosure 120 includes a first sub-enclosure 121 and a second sub-enclosure 122 extending in the first direction and spaced apart in the second direction, and a third sub-enclosure 123 and a fourth sub-enclosure 124 extending in the second direction and spaced apart in the first direction. The first sub-enclosure 121, the second sub-enclosure 122, the third sub-enclosure 123 and the fourth sub-enclosure 124 surround a rectangular receiving groove 140. The third sub-enclosure 123 is adjacent to the second sub-partition 133, the fourth sub-enclosure 124 is adjacent to the third sub-partition 134, a first installation groove 141 communicating with the first penetration hole 161 is formed between the third sub-enclosure 123 and the second sub-partition 133, and a second installation groove 142 communicating with the second penetration hole (not shown in the drawings) is formed between the fourth sub-enclosure 124 and the third sub-partition 134.

[0053] In some exemplary embodiments, Figure 2 As shown, the actuator 300 includes a first roller 310 installed in the first mounting groove 141 and a second roller 320 installed in the second mounting groove 142, and the elastic base membrane 200 includes a first end 201 and a second end 202 spaced apart along a first direction, the first end 201 is connected to the first roller 310, and the second end 202 is connected to the second roller 320.

[0054] In some exemplary embodiments, Figure 2As shown, the first roller 310 includes a first shaft 311 and a first clamping shaft 312 disposed at both ends of the first shaft 311, two first clamping seats 171 are disposed in the first installation groove 141, the first shaft 311 is located between the two first clamping seats 171, and the first clamping shaft 312 is seated in the corresponding first clamping seat 171, and the second roller 320 includes a second shaft 321 and a second clamping shaft 322 disposed at both ends of the second shaft 321, two second clamping seats 172 are disposed in the second installation groove 142, the second shaft 321 is located between the two second clamping seats 172, and the second clamping shaft 322 is seated in the corresponding second clamping seat 172. The first shaft 311 and the second shaft 321 are both cylindrical, the first clamping shaft 312 and the second clamping shaft 322 are also cylindrical, the first clamping shaft 312 is coaxially disposed with the first shaft 311, and the second clamping shaft 322 is coaxially disposed with the second shaft 321. The structure of the first roller 310 is the same as that of the second roller 320. The first roller 310 and the second roller 320 may be made of resin material.

[0055] In some exemplary embodiments, Figure 3a As shown, the first sub-partition 132 of the first partition 131 adjacent to the first sub-enclosure 121 is connected to the first sub-enclosure 121, and extends into the first mounting groove 141 to form a first extension portion 135, and extends into the second mounting groove 142 to form a second extension portion 136. One of the two first sockets 171 includes the first extension portion 135 and a first slot 173 arranged on the end surface of the first extension portion 135 away from the bottom plate 110. One of the two second sockets 172 includes the second extension portion 136 and a second slot 174 arranged on the end surface of the second extension portion 136 away from the bottom plate 110. The first card shaft 312 is embedded in the first card groove 173 and can rotate, and the second card shaft 322 is embedded in the second card groove 174 and can rotate.

[0056] In some exemplary embodiments, Figure 3aAs shown, the partition 130 also includes a second partition 137 disposed between the first partition 131 and the second sub-enclosure 122, and the second partition 137 can be connected to the second sub-enclosure 122. The second partition 137 forms a third penetration channel 164 communicating with the first penetration hole 161 with the second sub-partition 133 of the adjacent first partition 131, and forms a fourth penetration channel 165 communicating with the second penetration hole with the third sub-partition 134 of the adjacent first partition 131. The elastic base membrane 200 can be penetrated to the first penetration hole through the third penetration channel 164, and penetrated to the second penetration hole through the fourth penetration channel 165. The second partition 137 includes a third extension portion 138 extending into the first mounting groove 141 and a fourth extension portion 139 extending into the second mounting groove 142. The other of the two first sockets 171 includes a third extension portion 138 and a third slot 175 arranged on the end surface of the third extension portion 138 away from the bottom plate 110. The other of the two second sockets 172 includes a fourth extension portion 139 and a fourth slot 176 arranged on the end surface of the fourth extension portion 139 away from the bottom plate 110. The first card shaft 312 is embedded in the third slot 175 and can rotate. The second card shaft 322 is embedded in the fourth slot 176 and can rotate.

[0057] Figure 4 is a plan view of a first card slot of an exemplary embodiment of the present invention, Figure 5 FIG. 1 is a plan view of the second card slot of an exemplary embodiment of the present invention. In some exemplary embodiments, the first card slot 173 and the third card slot 175 have the same structure. Taking the first card slot 173 as an example, Figure 4 As shown, the first clamping slot 173 includes a first guide portion 1771 and a first clamping portion 1772 connected to the first guide portion 1771. The first guide portion 1771 is used to guide the first clamping shaft 312 into the first clamping portion 1772. The first clamping portion 1772 is used to support the first clamping shaft 312 and limit the movement of the first clamping shaft 312 in a direction perpendicular to the bottom plate 110. The second clamping slot 174 and the fourth clamping slot 176 have the same structure. Taking the second clamping slot 174 as an example, the second clamping slot 174 includes a second guide portion 1781 and a second clamping portion 1782 connected to the second guide portion 1781. The second guide portion 1781 is used to guide the second clamping shaft 322 into the second clamping portion 1782. The second clamping portion 1782 is used to support the second clamping shaft 322 and limit the movement of the second clamping shaft 322 in a direction perpendicular to the bottom plate 110. In this example, the first locking portion 1772 can limit the first locking shaft 312 from escaping from the first locking slot 173 and the third locking slot 175 during rotation, and the second locking portion 1782 can limit the second locking shaft 322 from escaping from the second locking slot 174 and the fourth locking slot 176 during rotation.

[0058] In some exemplary embodiments, Figure 4 and Figure 5As shown, the first guide portion 1771 and the second guide portion 1781 both include a guide center line perpendicular to the base plate 110, the first clamping portion 1772 and the second clamping portion 1782 are circular areas, the width of the first guide portion 1771 in the first direction is smaller than the diameter of the first clamping portion 1772, and the center of the first clamping portion 1772 is not on the straight line where the guide center line of the first guide portion 1771 is located, the width of the second guide portion 1781 in the first direction is smaller than the diameter of the second clamping portion 1782, and the center of the second clamping portion 1782 is not on the straight line where the guide center line of the second guide portion 1781 is located. Among them, the center of the first positioning portion 1772 of the first slot 173 can be closer to the second sub-partition 133 relative to the guide center line of the first guide portion 1771 of the first slot 173, the center of the first positioning portion 1772 of the third slot 175 can be closer to the second sub-partition 133 relative to the guide center line of the first guide portion 1771 of the third slot 175, the center of the second positioning portion 1782 of the second slot 174 can be closer to the third sub-partition 134 relative to the guide center line of the second guide portion 1781 of the second slot 174, and the center of the second positioning portion 1782 of the fourth slot 176 can be closer to the third sub-partition 134 relative to the guide center line of the second guide portion 1781 of the fourth slot 176.

[0059] Figure 6 This is a structural diagram of the third card slot position of an exemplary embodiment of the present invention. Figure 7 FIG. 1 is a partial enlarged view of the first roller of an exemplary embodiment of the present invention. In some exemplary embodiments, as Figure 3a As shown, a first shaft hole communicating with the first mounting groove 141 and a second shaft hole communicating with the second mounting groove 142 are provided on a surface of one of the first sub-enclosure 121 and the second sub-enclosure 122 away from the accommodating groove 140, and the actuator 300 includes a first drive shaft 330 passing through the first shaft hole and a second drive shaft 340 passing through the second shaft hole. Figure 6 and Figure 7As shown, a first matching structure 331 is provided at one end of the first driving shaft 330 adjacent to the first clamping shaft 312, a second matching structure 313 engaged with the first matching structure 331 is provided on the first clamping shaft 312 adjacent to the first driving shaft 330, the first driving shaft 330 is configured to be able to actuate the first roller 310 after the first matching structure 331 and the second matching structure 313 are engaged, a third matching structure (not shown in the drawings) is provided at one end of the second driving shaft 340 adjacent to the second clamping shaft, a fourth matching structure (not shown in the drawings) engaged with the third matching structure is provided on the second clamping shaft 322 adjacent to the second driving shaft 340, and the second driving shaft 340 is configured to actuate the second roller 320 after the third matching structure and the fourth matching structure are engaged. In one example, the first matching structure 331 can be one of a cross-shaped tooth or a cross-shaped groove engaged with the cross-shaped tooth, the second matching structure 313 can be the other of a cross-shaped tooth or a cross-shaped groove engaged with the cross-shaped tooth, the third matching structure can be one of a cross-shaped tooth or a cross-shaped groove engaged with the cross-shaped tooth, and the fourth matching structure can be the other of a cross-shaped tooth or a cross-shaped groove engaged with the cross-shaped tooth. Of course, the cross-shaped tooth can also be replaced with a straight tooth, a triangular tooth, or a hexagonal tooth, etc., and the cross-shaped groove can be replaced with a shape that matches the tooth.

[0060] In some exemplary embodiments, Figure 6 As shown, a first sealing ring 351 is provided on the first driving shaft 330, and the first sealing ring 351 is used to seal the side of the first shaft hole close to the first mounting groove 141. A second sealing ring is provided on the second driving shaft, and the second sealing ring is used to seal the side of the second shaft hole close to the second mounting groove. The first sealing ring 351 and the second sealing ring are both O-rings. The first sealing ring 351 and the second sealing ring can prevent bacteria and microorganisms from entering the culture chamber 150 from the first shaft hole and the second shaft hole, thereby ensuring the sterile environment of the culture chamber 150 and preventing cells from being contaminated.

[0061] In some exemplary embodiments, Figure 3a As shown, a first actuator 332 may be provided at one end of the first drive shaft 330 away from the first roller 310, and a second actuator 342 may be provided at one end of the second drive shaft 340 away from the second roller 320. The first actuator 332 may drive the first drive shaft 330 to actuate the first roller 310 to rotate, and the second actuator 342 may drive the second drive shaft 340 to actuate the second roller 320 to rotate. In one example, both the first actuator 332 and the second actuator 342 may be actuator handles, which manually control the rotation of the first roller 310 and the second roller 320. In another example, both the first actuator 332 and the second actuator 342 may be motors or cylinders, which are combined with a controller to automatically control the rotation of the first roller 310 and the second roller 320.

[0062] In some exemplary embodiments, Figure 4 and Figure 5 As shown, the other of the first sub-enclosure 121 and the second sub-enclosure 122 is provided with a fifth matching structure 125 corresponding to the position of the first mounting groove 141 and a sixth matching structure 126 corresponding to the position of the second mounting groove 142 on the surface facing the accommodating groove 140, the first clamping shaft 312 adjacent to the fifth matching structure 125 is provided with a seventh matching structure (not shown in the drawings) rotatably connected to the fifth matching structure 125, and the second clamping shaft 322 adjacent to the sixth matching structure 126 is provided with an eighth matching structure (not shown in the drawings) rotatably connected to the sixth matching structure 126. In one example, the fifth matching structure 125 can be one of the plug shaft and the socket, the seventh matching structure can be the other of the plug shaft or the socket, the sixth matching structure 126 can be one of the plug shaft and the socket, and the eighth matching structure can be the other of the plug shaft or the socket.

[0063] In some exemplary embodiments, the second matching structure 313 and the seventh matching structure have the same structure, which can be a cross slot, a straight tooth, a triangular tooth or a hexagonal tooth, etc. The fourth matching structure and the eighth matching structure have the same structure, which can be a cross slot, a straight tooth, a triangular tooth or a hexagonal tooth, etc.

[0064] In some exemplary embodiments, Figure 2 As shown, a fifth slot 314 is provided on the first roller 310, a sixth slot (not shown in the drawings) is provided on the second roller 320, a first clip 210 matching the fifth slot 314 is provided on the first end 201 of the elastic base membrane 200, and a second clip 220 matching the sixth slot is provided on the second end 202 of the elastic base membrane 200. The fifth slot 314 can be multiple and spaced along the second direction, the sixth slot can be multiple and spaced along the second direction, and the fifth slot 314 and the sixth slot are the same in number as the first partition 131. In this example, the second direction can be the extending direction of the first roller 310 and the second roller 320. The first clip 210 and the fifth slot 314 are detachably connected, and the second clip 220 and the sixth slot are detachably connected, so as to facilitate the replacement and removal of the elastic base membrane 200 for other experiments. The first clip 210 and the second clip 220 can both be straight clips, the fifth slot 314 is adapted to the first clip 210 , and the sixth slot is adapted to the second clip 220 . In the second direction, the lengths of the first clip 210 and the second clip 220 are both greater than the width of the elastic base membrane 200 .

[0065] Figure 8 for Figure 7 In some exemplary embodiments, as shown in FIG. Figure 7and Figure 8 As shown, a first limiting groove 315 is provided at the slot position of the fifth slot 314, and a second limiting groove is provided at the slot position of the sixth slot. In the second direction, the width of the fifth slot 314 is greater than the width of the first limiting groove 315, the width of the first limiting groove 315 is adapted to the width of the elastic base membrane 200, and is configured to limit the movement of the elastic base membrane 200 in the second direction. The width of the sixth slot is greater than the width of the second limiting groove, the width of the second limiting groove is adapted to the width of the elastic base membrane 200, and is configured to limit the movement of the elastic base membrane 200 in the second direction.

[0066] In some exemplary embodiments, Figure 2 As shown, the cell culture device 1 further includes a cover plate 400, which is disposed on the outer side of the enclosing plate 120 and covers the containing tank 140. The cover plate 400 includes a top plate 410 and a side plate 420 disposed on the circumference of the top plate 410, and the side plate 420 is located on a side of the enclosing plate 120 away from the containing tank 140. The cover plate 400 can be made of a transparent material to facilitate observation of the state of the cells in the culture chamber.

[0067] In some exemplary embodiments, Figure 2 As shown, the periphery of the bottom plate 110 protrudes out of the surrounding plate 120 in a direction away from the receiving groove 140 , and the side plate 420 abuts against the bottom plate 110 .

[0068] In some exemplary embodiments, Figure 2 As shown, the side plate 420 is provided with a first avoidance groove 421 for avoiding the first driving shaft 330 and a second avoidance groove 422 for avoiding the second driving shaft 340 .

[0069] In some exemplary embodiments, Figure 2 As shown, a liquid exchange port 430 is provided on the top plate 410, and the liquid exchange port 430 corresponds to the culture chamber 150. The liquid exchange port 430 includes a liquid inlet 431 and a liquid outlet 432. The liquid inlet 431 can be connected to the outlet of the liquid supply pump through a liquid supply pipeline, and the liquid outlet 432 can be connected to the inlet of the liquid discharge pump through a liquid discharge pipeline, and then it can be expanded into a small cell workstation to complete simple cell liquid replacement, drug screening and other tasks. In order to ensure the sterile environment in the culture chamber 150, a sealing ring can be provided at the liquid exchange port 430, and the sealing ring can seal the gap between the top plate 410 and the pipeline (liquid discharge pipeline and liquid supply pipeline).

[0070] In some exemplary embodiments, Figure 2 and 3aAs shown, the enclosure 120 is provided with a first anti-reverse structure 127, and the cover plate 400 is provided with a second anti-reverse structure 440 that cooperates with the first anti-reverse structure 127. The first anti-reverse structure 127 and the second anti-reverse structure 440 can prevent the first drive shaft 330 and the second drive shaft 340 from being damaged due to the reverse installation of the cover plate 400 and the culture dish 100, resulting in the first avoidance groove 421 not corresponding to the first drive shaft 330 and the second avoidance groove 422 not corresponding to the second drive shaft 340. The first anti-reverse structure 127 can be a first wedge angle provided on the first enclosure 120, and the second anti-reverse structure 440 can be a second wedge angle provided on the side plate 420.

[0071] In some exemplary embodiments, Figure 3a As shown, the distance between the surface of the partition 130 away from the bottom plate 110 and the bottom plate 110 is greater than the distance between the surface of the enclosure 120 away from the bottom plate 110 and the bottom plate 110. In one example, the cell culture device 1 further includes a first flange 1231 disposed on the third sub-enclosure 123 and a second flange 1241 disposed on the fourth sub-enclosure 124, the first flange 1231 and the second flange 1241 being disposed along the edge of the containing groove 140, and the distance between the surface of the first flange 1231 and the second flange 1241 away from the bottom plate 110 and the bottom plate 110 is equal to the distance between the surface of the partition 130 away from the bottom plate 110 and the bottom plate 110.

[0072] The technical solution of the cell culture device 1 according to the embodiment of the present invention is explained below by taking the working principle of the cell culture device 1 as an example.

[0073] In some exemplary embodiments, the number and position of the elastic base membrane 200 correspond to the first partition 131, the elastic base membrane 200 is penetrated into the corresponding first penetration hole 161 through the corresponding first penetration channel 162, the elastic base membrane 200 is penetrated into the corresponding second penetration hole through the corresponding second penetration channel 163, the elastic base membrane 200 covers the bottom of the culture chamber 150, the first clip 210 of the first end 201 of the elastic base membrane 200 is clamped into the fifth slot 314 corresponding to the first roller 310, and the second clip 220 of the second end 202 of the elastic base membrane 200 is clamped into the sixth slot corresponding to the second roller 320, thereby fixing the elastic base membrane 200. Then, the first clamping shaft 312 of the first roller 310 is clamped into the first clamping portion 1772 of the first clamping seat 171 through the first guide portion 1771 and is limited by the first clamping portion 1772 , and the second clamping shaft 322 of the second roller 320 is clamped into the second clamping portion 1782 of the second clamping seat 172 through the second guide portion 1781 and is limited by the second clamping portion 1782 . The first matching structure 331 on the first driving shaft 330 is clamped into the second matching structure 313 on the first clamping shaft 312 adjacent to the first driving shaft 330, the fifth matching structure 125 is rotatably connected to the seventh matching structure on the first clamping shaft 312 away from the first driving shaft 330, the third matching portion on the second driving shaft 340 is clamped into the fourth matching portion on the second clamping shaft 322 adjacent to the second driving shaft 340, the sixth matching structure 126 is rotatably connected to the eighth matching structure on the second clamping shaft 322 away from the second driving shaft 340, so that the first driving shaft 330 can actuate the first roller 310, the second driving shaft 340 can actuate the second roller 320, and when the first driving shaft 330 and the second driving shaft 340 rotate in opposite directions, the elastic base membrane 200 can be stretched or relaxed. During the installation process, in order to smoothly install the first roller 310 into the first installation groove 141 and the second roller 320 into the second installation groove 142, the first sub-enclosure 121 and the third sub-enclosure 122 can be pulled outward when installing the first roller 310 and the second roller 320, so that the first sub-enclosure 121 and the third sub-enclosure 122 are elastically deformed to expand the space of the first installation groove 141 and the second installation groove 142. Cover the cover plate 400, the first avoidance groove 421 on the cover plate 400 corresponds to the first drive shaft 330, and the second avoidance groove 422 on the cover plate 400 corresponds to the second drive shaft 340. Finally, insert the liquid supply pipeline into the liquid inlet 431 of the cover plate 400, insert the liquid discharge pipeline into the liquid outlet 432 of the cover plate 400, and the other ends of the liquid supply pipeline and the liquid discharge pipeline are connected to the corresponding pumps.

[0074] A certain amount of substances that promote cell adhesion, such as poly-lysine (PLL) or collagen, are pumped into the culture chamber 150. Then a certain amount of phosphate buffered saline (PBS) solution is pumped in to clean the residual liquid, and a cell suspension with a certain cell density is pumped in. The suspension is left to stand for more than 6 hours until the cells are attached to the elastic basement membrane 200, and then conventional cell culture can be performed.

[0075] Finally, according to the experimental requirements and in combination with Hu Ke's law, the first roller 310 is actuated to rotate by the first actuator 332, and the second roller 320 is actuated to rotate by the second actuator 342, so that the elastic basement membrane 200 is stretched to a predetermined length, thereby achieving a stretching effect on the cells, so as to simulate the growth environment of the cells in the body, for example Figure 9a and 9b As shown, the first actuator 332 actuates the first roller 310 to rotate clockwise, and the second actuator 342 actuates the second roller 320 to rotate counterclockwise, the elastic basement membrane 200 is stretched, and the cells attached to the elastic basement membrane 200 are stretched. Then, the first actuator 332 actuates the first roller 310 to rotate counterclockwise, and the second actuator 342 actuates the second roller 320 to rotate clockwise, the elastic basement membrane 200 relaxes from the stretched state to the original state, and the cells attached to the elastic basement membrane 200 also return to the static state. The above actions are repeated, so that the cells are in a dynamic environment.

[0076] It can be seen from the working principle of the cell culture device 1 of the exemplary embodiment of the present invention that the first actuator 332 and the second actuator 342 can actuate the first roller 310 and the second roller 320 to rotate in opposite directions. During the rotation of the first roller 310 and the second roller 320, the elastic basement membrane 200 can be stretched or relaxed, and the cells attached to the elastic basement membrane 200 can move synchronously with the elastic basement membrane 200, thereby realizing cell culture in a dynamic environment, thereby more realistically reflecting the state of cell culture in the body.

[0077] Although the embodiments disclosed in the present invention are as above, the contents described are only embodiments adopted to facilitate understanding of the present invention and are not intended to limit the present invention. Any technician in the field to which the present invention belongs can make any modifications and changes in the form and details of implementation without departing from the spirit and scope disclosed in the present invention, but the patent protection scope of the present invention shall still be defined by the attached claims.

Claims

1. A cell culture device, characterized in that: include: A culture dish, comprising a bottom plate, a surrounding plate and at least one partition plate, wherein the surrounding plate is arranged along the circumference of the bottom plate, the bottom plate and the surrounding plate form a receiving groove, the partition plate is arranged in the receiving groove and defines at least one culture chamber, and a through hole is provided between the partition plate and the bottom plate and communicated with the culture chamber; An elastic basement membrane is penetrated through the penetration hole and covers the position of the bottom plate corresponding to the culture chamber; an actuator, at least partially disposed in the receiving groove and located between the enclosure plate and the partition plate, connected to the end of the elastic base membrane, and used to stretch the elastic base membrane in a direction parallel to the bottom plate; The partition includes a first partition, the first partition includes a first sub-partition extending along a first direction, and a second sub-partition and a third sub-partition arranged on the first sub-partition at intervals along the first direction and extending along a second direction, the first sub-partition is arranged on the bottom plate, there is a gap between the second sub-partition and the bottom plate, there is a gap between the third sub-partition and the bottom plate, the through holes include a first through hole between the second sub-partition and the bottom plate, and a second through hole between the third sub-partition and the bottom plate, and the first direction and the second direction intersect; The first partitions include a plurality of partitions spaced apart along the second direction; the number of the elastic base membranes is the same as the number of the first partitions, and the positions thereof correspond one to one; The enclosure includes a first sub-enclosure and a second sub-enclosure extending in the first direction and spaced apart in the second direction, and a third sub-enclosure and a fourth sub-enclosure extending in the second direction and spaced apart in the first direction, the third sub-enclosure is adjacent to the second sub-enclosure, the fourth sub-enclosure is adjacent to the third sub-enclosure, a first mounting groove communicating with the first through-hole is formed between the third sub-enclosure and the second sub-enclosure, and a second mounting groove communicating with the second through-hole is formed between the fourth sub-enclosure and the third sub-enclosure; The actuator includes a first roller installed in the first mounting groove and a second roller installed in the second mounting groove. The elastic base membrane includes a first end and a second end spaced apart along a first direction. The first end is connected to the first roller, and the second end is connected to the second roller.

2. The cell culture device according to claim 1, characterized in that: In a direction perpendicular to the bottom plate, the diameter of the through hole is adapted to the thickness of the elastic base membrane, and in the second direction, the diameter of the through hole is adapted to the width of the elastic base membrane.

3. The cell culture device according to claim 1, characterized in that: The first roller includes a first shaft body and a first clamping shaft arranged at both ends of the first shaft body, two first clamping seats are arranged in the first installation groove, the first shaft body is located between the two first clamping seats, and the first clamping shaft is seated in the corresponding first clamping seat, the second roller includes a second shaft body and a second clamping shaft arranged at both ends of the second shaft body, two second clamping seats are arranged in the second installation groove, the second shaft body is located between the two second clamping seats, and the second clamping shaft is seated in the corresponding second clamping seat.

4. The cell culture device according to claim 3, characterized in that: The first sub-partition of the first partition adjacent to the first sub-enclosure extends into the first mounting groove to form a first extension portion, and extends into the second mounting groove to form a second extension portion, one of the two first sockets includes the first extension portion and a first socket provided on the end surface of the first extension portion away from the bottom plate, one of the two second sockets includes the second extension portion and a second socket provided on the end surface of the second extension portion away from the bottom plate, the first socket shaft is embedded in the first socket shaft and can rotate, and the second socket shaft is embedded in the second socket shaft and can rotate.

5. The cell culture device according to claim 4, characterized in that: The partition also includes a second partition arranged between the first partition and the second sub-enclosure, the second partition includes a third extension portion extending into the first mounting groove and a fourth extension portion extending into the second mounting groove, the other of the two first sockets includes the third extension portion and a third slot arranged on the end surface of the third extension portion away from the bottom plate, the other of the two second sockets includes the fourth extension portion and a fourth slot arranged on the end surface of the fourth extension portion away from the bottom plate, the first card shaft is embedded in the third card slot and can rotate, and the second card shaft is embedded in the fourth card slot and can rotate.

6. The cell culture device according to claim 3, characterized in that: The first and second sub-enclosures are provided with a first axial hole connected to the first mounting groove and a second axial hole connected to the second mounting groove on a surface of one of the first sub-enclosure and the second sub-enclosure away from the accommodating groove, and the actuator includes a first driving shaft passed through the first axial hole and a second driving shaft passed through the second axial hole, a first matching structure is provided at one end of the first driving shaft adjacent to the first clamping shaft, a second matching structure engaged with the first matching structure is provided on the first clamping shaft adjacent to the first driving shaft, the first driving shaft is configured to be able to actuate the first roller after the first matching structure and the second matching structure are engaged, a third matching structure is provided at one end of the second driving shaft adjacent to the second clamping shaft, a fourth matching structure engaged with the third matching structure is provided on the second clamping shaft adjacent to the second driving shaft, and the second driving shaft is configured to actuate the second roller after the third matching structure and the fourth matching structure are engaged.

7. The cell culture device according to claim 6, characterized in that: A first actuator is provided at one end of the first drive shaft away from the first roller, and a second actuator is provided at one end of the second drive shaft away from the second roller. The first actuator is configured to drive the first drive shaft to actuate the first roller to rotate, and the second actuator is configured to drive the second drive shaft to actuate the second roller to rotate.

8. The cell culture device according to claim 6, characterized in that: A fifth matching structure corresponding to the position of the first installation groove and a sixth matching structure corresponding to the position of the second installation groove are arranged on the surface of the other of the first sub-enclosure plate and the second sub-enclosure plate facing the accommodating groove, a seventh matching structure rotatably connected to the fifth matching structure is arranged on the first clamping shaft adjacent to the fifth matching structure, and an eighth matching structure rotatably connected to the sixth matching structure is arranged on the second clamping shaft adjacent to the sixth matching structure.

9. The cell culture device according to claim 1, characterized in that: The first roller is provided with a fifth card slot, the second roller is provided with a sixth card slot, the first end of the elastic base membrane is provided with a first card clip that cooperates with the fifth card slot, and the second end of the elastic base membrane is provided with a second card clip that cooperates with the sixth card slot.

10. The cell culture device according to claim 9, characterized in that: A first limiting groove is provided at the slot position of the fifth slot, and a second limiting groove is provided at the slot position of the sixth slot. In the second direction, the width of the fifth slot is greater than the width of the first limiting groove, the width of the first limiting groove is adapted to the width of the elastic base membrane, and is configured to limit the movement of the elastic base membrane in the second direction. The width of the sixth slot is greater than the width of the second limiting groove, the width of the second limiting groove is adapted to the width of the elastic base membrane, and is configured to limit the movement of the elastic base membrane in the second direction.

11. The cell culture device according to claim 1, characterized in that: The cell culture device also includes a cover plate, which is arranged on the outer side of the surrounding plate and covers the containing tank. The cover plate includes a top plate and a side plate arranged on the circumference of the top plate, and the side plate is located on the side of the surrounding plate away from the containing tank. A liquid exchange port is arranged on the top plate, and the liquid exchange port corresponds to the culture chamber.

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