CN114381372A

CN114381372A - Cell culture device

Info

Publication number: CN114381372A
Application number: CN202011132449.5A
Authority: CN
Inventors: 范蓓媛; 徐为峰; 王嘉鹏; 丁丁
Original assignee: BOE Technology Group Co Ltd
Current assignee: BOE Technology Group Co Ltd
Priority date: 2020-10-21
Filing date: 2020-10-21
Publication date: 2022-04-22

Abstract

This paper discloses a cell culture device, relates to the biotechnology field, includes: the culture dish comprises a bottom plate, enclosing plates and at least one partition plate, wherein the enclosing plates are arranged along the circumferential direction of the bottom plate, the bottom plate and the enclosing plates enclose an accommodating groove, the partition plate is arranged in the accommodating groove and limits at least one culture chamber, and a through hole communicated with the culture chamber is formed between the partition plate and the bottom plate; the elastic basement membrane penetrates through the through holes and covers the position, corresponding to the culture chamber, of the bottom plate; and the actuating mechanism is at least partially arranged in the accommodating groove, positioned between the enclosing plate and the partition plate and used for stretching the elastic base film. The elastic basement membrane is arranged at the bottom of the culture chamber and can be stretched under the driving of the actuating mechanism, and cells on the elastic basement membrane can be stretched synchronously in the stretching process of the elastic basement membrane, so that the growth environment of the cells in vivo can be simulated, and the state of the cells cultured in vivo is really reflected.

Description

Cell culture device

Technical Field

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

Background

Cell culture is the basic unit operation of biological research, and the micro-fluidic chip builds a two-dimensional or three-dimensional network structure at a micron level or even a nanometer level by relying on a micro-processing technology, and can be integrated with functional units such as a micro pump, a micro valve, a micro reactor and the like. Microfluidic cell culture has the characteristics of reducing sample amount, customizing and researching various microenvironments in the same culture device, and the like.

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 of hydrostatic pressure, shear force generated during blood flow and tensile force generated during vasomotor process, however, in the conventional in vitro cell culture, the cells are in a relatively static environment, which is very different from the in vivo environment, and the in vitro cell culture cannot truly reflect the state of the cells in vivo.

Disclosure of Invention

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.

The embodiment of the invention provides a cell culture device which can simulate the growth environment of cells in vivo and truly reflect the state of the cells cultured in vivo.

The cell culture apparatus provided by the embodiment of the invention comprises:

the culture dish comprises a bottom plate, enclosing plates and at least one partition plate, wherein the enclosing plates are arranged along the circumferential direction of the bottom plate, the bottom plate and the enclosing plates enclose an accommodating groove, the partition plate is arranged in the accommodating groove and limits at least one culture chamber, and a through hole communicated with the culture chamber is formed between the partition plate and the bottom plate;

the elastic basement membrane penetrates through the through holes and covers the position, corresponding to the culture chamber, of the bottom plate;

and the actuating mechanism is at least partially arranged in the accommodating groove, positioned between the enclosing plate and the partition plate and used for stretching the elastic base film.

In some exemplary embodiments, the partition plate includes a first partition plate including a first sub-partition plate extending in a first direction, and a second sub-partition plate and a third sub-partition plate spaced apart from each other in the first direction and extending in a second direction, the first sub-partition plate being disposed on the bottom plate, the second sub-partition plate having a space from the bottom plate, the third sub-partition plate having a space from the bottom plate, the through holes including a first through hole between the second sub-partition plate and the bottom plate and a second through hole between the third sub-partition plate and the bottom plate, the first direction intersecting the second direction.

In some exemplary embodiments, the aperture of the through-holes is adapted to the thickness of the elastic base film in a direction perpendicular to the base plate, and the aperture of the through-holes is adapted to the width of the elastic base film in the second direction.

In some exemplary embodiments, the first barrier includes a plurality of partitions disposed at intervals in the second direction.

In some exemplary embodiments, the enclosing plates include a first sub enclosing plate and a second sub enclosing plate which extend along the first direction and are arranged at intervals along the second direction, and a third sub enclosing plate and a fourth sub enclosing plate which extend along the second direction and are arranged at intervals along the first direction, the third sub enclosing plate is adjacent to the second sub partition plate, the fourth sub enclosing plate is adjacent to the third sub partition plate, a first installation groove communicated with the first through hole is formed between the third sub enclosing plate and the second sub partition plate, and a second installation groove communicated with the second through hole is formed between the fourth sub enclosing plate and the third sub partition plate.

In some exemplary embodiments, the actuator includes a first roller installed in the first installation groove and a second roller installed in the second installation groove, and the elastic base film includes a first end and a second end spaced apart in the first direction, the first end being connected to the first roller, and the second end being connected to the second roller.

In some exemplary embodiments, the first roller includes a first shaft body and first clamping shafts disposed at two ends of the first shaft body, two first clamping seats are disposed in the first mounting groove, the first shaft body is disposed between the two first clamping seats, the first clamping shafts are seated in the corresponding first clamping seats, the second roller includes a second shaft body and second clamping shafts disposed at two ends of the second shaft body, two second clamping seats are disposed in the second mounting groove, the second shaft body is disposed between the two second clamping seats, and the second clamping shafts are seated in the corresponding second clamping seats.

In some exemplary embodiments, a first sub-partition plate of a first partition plate adjacent to the first sub-enclosure plate extends into the first mounting groove to form a first extending portion, and extends into the second mounting groove to form a second extending portion, one of the two first clamping seats comprises the first extending portion and a first clamping groove arranged on an end face, far away from the bottom plate, of the first extending portion, one of the two second clamping seats comprises the second extending portion and a second clamping groove arranged on an end face, far away from the bottom plate, of the second extending portion, the first clamping shaft is embedded in the first clamping groove and can rotate, and the second clamping shaft is embedded in the second clamping groove and can rotate.

In some exemplary embodiments, the partition board further includes a second partition board disposed between the first partition board and the second sub enclosure board, the second partition board includes a third extending portion extending into the first mounting groove and a fourth extending portion extending into the second mounting groove, another of the two first clamping seats includes a third extending portion and a third clamping groove disposed on an end surface of the third extending portion away from the bottom plate, another of the two second clamping seats includes a fourth extending portion and a fourth clamping groove disposed on an end surface of the fourth extending portion away from the bottom plate, the first clamping shaft is embedded in the third clamping groove and can rotate, and the second clamping shaft is embedded in the fourth clamping groove and can rotate.

In some exemplary embodiments, a first shaft hole communicated with the first mounting groove and a second shaft hole communicated with the second mounting groove are formed in a surface, away from the receiving groove, of one of the first sub surrounding plate and the second sub surrounding plate, the actuator includes a first driving shaft penetrating through the first shaft hole and a second driving shaft penetrating through the second shaft hole, a first matching structure is arranged at one end, adjacent to the first clamping shaft, of the first driving shaft, a second matching structure clamped with the first matching structure is arranged on the first clamping shaft, the first driving shaft is arranged to actuate the first roller after the first matching structure and the second matching structure are clamped, a third matching structure is arranged at one end, adjacent to the second clamping shaft, of the second driving shaft, a fourth matching structure clamped with the third matching structure is arranged on the second clamping shaft, and the second driving shaft is arranged to actuate the second roller after the third matching structure and the fourth matching structure are clamped.

In some exemplary embodiments, an end of the first drive shaft remote from the first roller is provided with a first actuator, an end of the second drive shaft remote from the second roller is provided with a second actuator, 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.

In some exemplary embodiments, a fifth matching structure corresponding to the first mounting groove and a sixth matching structure corresponding to the second mounting groove are arranged on the surface of the other of the first sub enclosing plate and the second sub enclosing plate facing the receiving groove, a seventh matching structure rotatably connected with the fifth matching structure is arranged on the first clamping shaft adjacent to the fifth matching structure, and an eighth matching structure rotatably connected with the sixth matching structure is arranged on the second clamping shaft adjacent to the sixth matching structure.

In some exemplary embodiments, a fifth locking groove is disposed on the first roller, a sixth locking groove is disposed on the second roller, a first end of the elastic basement membrane is provided with a first locking clip cooperating with the fifth locking groove, and a second end of the elastic basement membrane is provided with a second locking clip cooperating with the sixth locking groove.

In some exemplary embodiments, a notch position of the fifth card slot is provided with a first limiting groove, a notch position of the sixth card slot is provided with a second limiting groove, in the second direction, a width of the fifth card slot is greater than a width of the first limiting groove, the width of the first limiting groove is adapted to the width of the elastic base film and is configured to limit the movement of the elastic base film in the second direction, the width of the sixth card slot is greater than a width of the second limiting groove, the width of the second limiting groove is adapted to the width of the elastic base film and is configured to limit the movement of the elastic base film in the second direction.

In some exemplary embodiments, the cell culture apparatus further includes a cover plate covering the outer side of the enclosing plate and covering the containing groove, the cover plate includes a top plate and a side plate disposed in the circumferential direction of the top plate, the side plate is located on the side of the enclosing plate away from the containing groove, the top plate is provided with a liquid exchange port, and the liquid exchange port corresponds to the culture chamber.

The embodiment of the invention provides a cell culture device, wherein the bottom of a culture chamber is provided with an elastic basement membrane, the elastic basement membrane can be stretched under the driving of an actuating mechanism, and cells on the elastic basement membrane can be stretched in the stretching process of the elastic basement membrane, so that the growth environment of the cells in vivo can be simulated, and the state of the cells cultured in vivo can be truly reflected.

Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.

Other aspects will be apparent upon reading and understanding the attached drawings and detailed description.

Drawings

The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the example serve to explain the principles of the invention and not to limit the invention.

FIG. 1 is a perspective view of a cell culture apparatus according to an exemplary embodiment of the present invention;

FIG. 2 is an exploded view of a cell culture apparatus according to an exemplary embodiment of the present invention;

FIG. 3a is a block diagram of a culture dish according to an exemplary embodiment of the present invention;

FIG. 3b is a cross-sectional view taken at the location A-A in FIG. 3 a;

FIG. 4 is a plan view of a first card slot in accordance with an exemplary embodiment of the present invention;

FIG. 5 is a plan view of a second card slot in accordance with an exemplary embodiment of the present invention;

FIG. 6 is a block diagram of a third card slot location in accordance with an exemplary embodiment of the present invention;

FIG. 7 is an enlarged partial view of a first roller in accordance with an exemplary embodiment of the present invention;

FIG. 8 is a cross-sectional view taken at the location A-A in FIG. 7;

FIG. 9a is a schematic view of an elastic basement membrane of a cell culture apparatus according to an exemplary embodiment of the invention in an unstretched state;

FIG. 9b is a schematic view of the elastic basement membrane of the cell culture apparatus of an exemplary embodiment of the invention in a stretched state.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be noted that the embodiments and features of the embodiments in the present application may be arbitrarily combined with each other without conflict.

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", "opposite", "four corners", "periphery", "mouth" structure ", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of describing the present invention and simplifying the description, but do not indicate or imply that the structures referred to have specific orientations, are configured and operated in specific orientations, and thus, are not to be construed as limiting the present invention.

In the description of the embodiments of the present invention, unless otherwise explicitly specified or limited, the terms "connected," "directly connected," "indirectly connected," "fixedly connected," "mounted," and "assembled" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; the terms "mounted," "connected," and "fixedly connected" may be directly connected or indirectly connected through intervening media, or may be connected through two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

The embodiment of the invention provides a cell culture device, wherein the bottom of a culture chamber is provided with an elastic basement membrane, the elastic basement membrane can be stretched under the driving of an actuating mechanism, and cells attached to the elastic basement membrane can be simultaneously stretched in the stretching process of the elastic basement membrane, so that the growth environment of the cells in vivo can be simulated, and the state of the cells cultured in vivo can be truly reflected.

The technical solution of the exemplary embodiment of the present invention is exemplarily described below with reference to the accompanying drawings.

FIG. 1 is a perspective view of a cell culture apparatus according to an exemplary embodiment of the present invention, and FIG. 2 is an exploded view of the cell culture apparatus according to the exemplary embodiment of the present invention. In some exemplary embodiments, the cell culture apparatus 1 is used for in vitro culture of cells and provides a tensile force to the cells during cell culture, simulating the growth environment of the cells in vivo. Therefore, the cell culture apparatus 1 mainly includes a culture dish 100, an elastic base 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 comprises a bottom plate 110, a surrounding plate 120 and at least one partition 130, wherein the surrounding plate 120 is arranged along the circumference of the bottom plate 110, the bottom plate 110 and the surrounding plate 120 define a containing groove 140, the partition 130 is arranged in the containing groove 140 and defines at least one culture chamber 150, a through hole communicated with the culture chamber 150 is formed between the partition 130 and the bottom plate 110, 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 a certain tensile property, cells can be attached to the elastic basement membrane 200, and the elastic basement membrane 200 can be made of Polydimethylsiloxane (PDMS), which has biocompatibility and good elasticity and can be used for the cells to attach and grow on. The elastic base membrane 200 is perforated and covers the bottom plate 110 at a position corresponding to the culture chamber 150. The actuator 300 is at least partially disposed in the receiving groove 140 between the enclosing plate 120 and the partition 130, and the actuator 300 is connected to an end of the elastic base film 200 and serves to stretch the elastic base film 200. When the actuator 300 stretches the elastic basement membrane 200, the cells attached to the elastic basement membrane 200 will stretch simultaneously, thereby simulating the stretching force generated during the vasomotor process of the cells in vivo. According to hooke's law, after a solid material is stressed, the stress and strain of the material are in a linear relationship, i.e., F ═ -k · x, where k is an elastic coefficient and is only related to the material properties, x is a deformation length, and F is an elastic force, therefore, the actuator 300 stretches the elastic basement membrane 200 to a specific length, so as to generate a specific elastic force, and the cells are subjected to a specific stretching force on the elastic basement membrane 200.

The invention exemplarily provides a cell culture apparatus 1, by arranging an elastic basement membrane 200 at the bottom of a culture chamber 150, the elastic basement membrane 200 can be stretched under the driving of an actuating mechanism 300, and cells attached to the elastic basement membrane 200 can be simultaneously stretched in the process of stretching the elastic basement membrane 200, so that the growth environment of the cells in vivo can be simulated, and the state of the cells cultured in vivo can be truly reflected.

FIG. 3a is a block diagram of a culture dish 100 according to an exemplary embodiment of the present invention, and FIG. 3b is a cross-sectional view taken at a position A-A in FIG. 3 a. In some exemplary embodiments, as shown in fig. 3a and 3b, the barrier 130 includes a first barrier 131, the first barrier 131 includes a first sub-barrier 132 extending in a first direction, and a second sub-barrier 133 and a third sub-barrier 134 spaced apart from each other in the first direction on the first sub-barrier 132 and extending in a second direction, the first sub-barrier 132 is disposed on the bottom plate 110, the second sub-barrier 133 is spaced apart from the bottom plate 110, the third sub-barrier 134 is spaced apart from the bottom plate 110, the through holes 160 include a first through hole 161 between the second sub-barrier 133 and the bottom plate 110 and a second through hole (not shown in the drawings) between the third sub-barrier 134 and the bottom plate 110, the first direction and the second direction intersect, the first direction is an X direction as shown in fig. 3a, the second direction is a Y direction as shown in fig. 3a, the first direction and the second direction intersect including the first direction and the second direction are perpendicular, the perpendicular may here be substantially perpendicular, and the angle between the first direction and the second direction may be 87 °, 90 ° or 93 °. The aperture of the through hole 160 is adapted to the thickness of the elastic base film 200 in a direction perpendicular to the base plate 110, and the aperture of the through hole 160 is adapted to the width of the elastic base film 200 in the second direction. The adaptation here can be understood as: the aperture diameter of the through-holes 160 is equal to or slightly larger than the thickness of the elastic base film 200 in a direction perpendicular to the base plate 110, and slightly larger here may be that the aperture diameter of the through-holes 160 is 1.01 to 1.2 times the thickness of the elastic base film 200. In the second direction, the aperture diameter of the through-holes 160 is equal to or slightly larger than the width of the elastic base film 200, where slightly larger may be that the aperture diameter of the through-holes 160 is 1.01 to 1.2 times the width of the elastic base film 200. The through holes 160 may be referred to as a first through hole 161 and a second through hole.

In some exemplary embodiments, as shown in fig. 3a and 3b, the first barrier 131 includes a plurality disposed at intervals in the second direction. The number of the elastic base film 200 may be the same as that of the first partitions 131 and the positions correspond one to one. The two first partition plates 131 adjacent in the second direction may be defined as a first front partition plate 131a and a first rear partition plate 131b, the second sub-partition plate 133 and the third sub-partition plate 134 of the first front partition plate 131a are adjacent to the first sub-partition plate 132 of the first rear partition plate 131b, wherein a first through passage 162 communicating with the first through hole 161 is formed between the second sub-partition plate 133 of the first front partition plate 131a and the first sub-partition plate 132 of the first rear partition plate 131b, and a second through passage 163 communicating with the second through hole (not shown in the drawings) is formed between the third sub-partition plate 134 of the first front partition plate 131a and the first sub-partition plate 132 of the first rear partition plate 131 b. The elastic base film 200 may be formed through the first through hole 161 via the first through channel 162 and the second through hole via the second through channel 163.

In some exemplary embodiments, as shown in fig. 3a, the enclosing plate 120 includes a first sub enclosing plate 121 and a second sub enclosing plate 122 extending along the first direction and spaced apart along the second direction, and a third sub enclosing plate 123 and a fourth sub enclosing plate 124 extending along the second direction and spaced apart along the first direction, and the first sub enclosing plate 121, the second sub enclosing plate 122, the third sub enclosing plate 123 and the fourth sub enclosing plate 124 enclose a rectangular accommodating groove 140. The third sub surrounding plate 123 is adjacent to the second sub partition 133, the fourth sub surrounding plate 124 is adjacent to the third sub partition 134, a first mounting groove 141 communicated with the first through hole 161 is formed between the third sub surrounding plate 123 and the second sub partition 133, and a second mounting groove 142 communicated with the second through hole (not shown in the figure) is formed between the fourth sub surrounding plate 124 and the third sub partition 134.

In some exemplary embodiments, as shown in fig. 2, 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 film 200 includes a first end 201 and a second end 202 spaced apart in a first direction, the first end 201 being connected to the first roller 310, and the second end 202 being connected to the second roller 320.

In some exemplary embodiments, as shown in fig. 2, the first roller 310 includes a first shaft 311 and first clamping shafts 312 disposed at two ends of the first shaft 311, two first clamping seats 171 are disposed in the first mounting groove 141, the first shaft 311 is located between the two first clamping seats 171, the first clamping shafts 312 are seated in the corresponding first clamping seats 171, the second roller 320 includes a second shaft 321 and second clamping shafts 322 disposed at two ends of the second shaft 321, two second clamping seats 172 are disposed in the second mounting groove 142, the second shaft 321 is located between the two second clamping seats 172, and the second clamping shafts 322 are seated in the corresponding second clamping seats 172. The first shaft body 311 and the second shaft body 321 are both cylindrical, the first clamping shaft 312 and the second clamping shaft 322 are also cylindrical, the first clamping shaft 312 and the first shaft body 311 are coaxially arranged, and the second clamping shaft 322 and the second shaft body 321 are coaxially arranged. 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 a resin material.

In some exemplary embodiments, as shown in fig. 3a, the first sub dividing plate 132 of the first dividing plate 131 adjacent to the first sub surrounding plate 121 is connected to the first sub surrounding plate 121, and extends into the first installation groove 141 to form a first extending portion 135, and extends into the second installation groove 142 to form a second extending portion 136, one of the two first clamping seats 171 includes the first extending portion 135 and a first clamping groove 173 disposed on an end surface of the first extending portion 135 away from the bottom plate 110, one of the two second clamping seats 172 includes the second extending portion 136 and a second clamping groove 174 disposed on an end surface of the second extending portion 136 away from the bottom plate 110, the first clamping shaft 312 is embedded in the first clamping groove 173 and can rotate, and the second clamping shaft 322 is embedded in the second clamping groove 174 and can rotate.

In some exemplary embodiments, as shown in fig. 3a, the partition 130 further includes a second partition 137 disposed between the first partition 131 and the second sub-enclosing plate 122, and the second partition 137 may be connected to the second sub-enclosing plate 122. The second partition plate 137 and the second sub-partition plate 133 of the adjacent first partition plate 131 form a third through passage 164 communicating with the first through hole 161, and the third sub-partition plate 134 of the adjacent first partition plate 131 forms a fourth through passage 165 communicating with the second through hole. The elastic base film 200 may be formed through the third through-channel 164 to the first through-hole and through the fourth through-channel 165 to the second through-hole. The second partition 137 includes a third extending portion 138 extending into the first mounting groove 141 and a fourth extending portion 139 extending into the second mounting groove 142, the other of the two first clamping seats 171 includes the third extending portion 138 and a third clamping groove 175 disposed on an end surface of the third extending portion 138 away from the bottom plate 110, the other of the two second clamping seats 172 includes the fourth extending portion 139 and a fourth clamping groove 176 disposed on an end surface of the fourth extending portion 139 away from the bottom plate 110, the first clamping shaft 312 is embedded in the third clamping groove 175 and can rotate, and the second clamping shaft 322 is embedded in the fourth clamping groove 176 and can rotate.

Fig. 4 is a plan view of a first card slot in accordance with an exemplary embodiment of the present invention, and fig. 5 is a plan view of a second card slot in accordance with an exemplary embodiment of the present invention. In some exemplary embodiments, the first latch recess 173 and the third latch recess 175 are identical in structure, and taking the first latch recess 173 as an example, as shown in fig. 4, the first latch recess 173 includes a first guide portion 1771 and a first latch portion 1772 communicating with the first guide portion 1771, the first guide portion 1771 is used for guiding the first latch shaft 312 into the first latch portion 1772, and the first latch portion 1772 is used for supporting the first latch shaft 312 and limiting movement of the first latch shaft 312 in a direction perpendicular to the bottom plate 110. The second card slot 174 and the fourth card slot 176 have the same structure, and taking the second card slot 174 as an example, the second card slot 174 includes a second guiding portion 1781 and a second blocking portion 1782 communicated with the second guiding portion 1781, the second guiding portion 1781 is used for guiding the second card shaft 322 into the second blocking portion 1782, and the second blocking portion 1782 is used for supporting the second card shaft 322 and limiting the movement of the second card shaft 322 in the direction perpendicular to the bottom plate 110. In this example, the first detent 1772 may limit the first latch shaft 312 from being disengaged from the first and third latch recesses 173 and 175 during rotation, and the second detent 1782 may limit the second latch shaft 322 from being disengaged from the second and fourth latch recesses 174 and 176 during rotation.

In some exemplary embodiments, as shown in fig. 4 and 5, each of the first and

second guiding portions

1771 and 1781 includes a guiding center line perpendicular to the bottom plate 110, the first and

second detent portions

1772 and 1782 are circular areas, a width of the first guiding portion 1771 in the first direction is smaller than a diameter of the first detent portion 1772, a center of the first detent portion 1772 is not on a straight line of the guiding center line of the first guiding portion 1771, a width of the second guiding portion 1781 in the first direction is smaller than a diameter of the second detent portion 1782, and a center of the second detent portion 1782 is not on a straight line of the guiding center line of the second guiding portion 1781. The center of the first latching portion 1772 of the first latching recess 173 may be closer to the second sub-spacer 133 than the guiding center line of the first guiding portion 1771 of the first latching recess 173, the center of the first latching portion 1772 of the third latching recess 175 may be closer to the second sub-spacer 133 than the guiding center line of the first guiding portion 1771 of the third latching recess 175, the center of the second latching portion 1782 of the second latching recess 174 may be closer to the third sub-spacer 134 than the guiding center line of the second guiding portion 1781 of the second latching recess 174, and the center of the second latching portion 1782 of the fourth latching recess 176 may be closer to the third sub-spacer 134 than the guiding center line of the second guiding portion 1781 of the fourth latching recess 176.

Fig. 6 is a structural view of a position of a third slot according to an exemplary embodiment of the present invention, and fig. 7 is a partially enlarged view of a first roller according to an exemplary embodiment of the present invention. In some exemplary embodiments, as shown in fig. 3a, a surface of one of the first and second

sub enclosing plates

121 and 122, which is away from the receiving groove 140, is provided with a first shaft hole communicating with the first receiving groove 141 and a second shaft hole communicating with the second receiving groove 142, and the actuator 300 includes a first driving shaft 330 penetrating the first shaft hole and a second driving shaft 340 penetrating the second shaft hole. As shown in fig. 6 and 7, a first mating structure 331 is disposed at an end of the first driving shaft 330 adjacent to the first locking shaft 312, a second mating structure 313 engaged with the first mating structure 331 is disposed on the first locking shaft 312 adjacent to the first driving shaft 330, the first driving shaft 330 is configured to actuate the first roller 310 after the first mating structure 331 and the second mating structure 313 are engaged, a third mating structure (not shown in the drawings) is disposed at an end of the second driving shaft 340 adjacent to the second locking shaft, a fourth mating structure (not shown in the drawings) engaged with the third mating structure is disposed on the second locking 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 mating structure and the fourth mating structure are engaged. In an example, the first mating structure 331 may be one of a cross latch or a cross slot engaged with the cross latch, the second mating structure 313 may be the other of the cross latch or the cross slot engaged with the cross latch, the third mating structure may be one of the cross latch or the cross slot engaged with the cross latch, and the fourth mating structure may be the other of the cross latch or the cross slot engaged with the cross latch. Of course, the cross latch can be replaced by a straight latch, a triangular latch or a hexagonal latch, and the cross slot can be replaced by a shape matched with the latch.

In some exemplary embodiments, as shown in fig. 6, a first sealing ring 351 is disposed on the first driving shaft 330, and the first sealing ring 351 is used for sealing a side of the first shaft hole close to the first mounting groove 141. And a second sealing ring is arranged on the second driving shaft and used for sealing one side of the second shaft hole close to the second mounting groove. The first seal ring 351 and the second seal ring are both O-ring seals. The first sealing ring 351 and the second sealing ring can prevent bacteria, microorganisms and the like from entering the culture chamber 150 from the first shaft hole and the second shaft hole, so that the sterile environment of the culture chamber 150 is ensured, and the cells are prevented from being polluted.

In some exemplary embodiments, as shown in fig. 3a, an end of the first driving shaft 330 remote from the first roller 310 may be provided with a first actuating member 332, an end of the second driving shaft 340 remote from the second roller 320 may be provided with a second actuating member 342, the first actuating member 332 may drive the first driving shaft 330 to actuate the first roller 310 to rotate, and the second actuating member 342 may drive the second driving shaft 340 to actuate the second roller 320 to rotate. In one example, the first actuator 332 and the second actuator 342 can each be an actuation handle that manually controls the rotation of the first roller 310 and the second roller 320. In another example, the first actuator 332 and the second actuator 342 can be motors or cylinders, and in combination with a controller, the first roller 310 and the second roller 320 are automatically controlled to rotate.

In some exemplary embodiments, as shown in fig. 4 and 5, a fifth mating structure 125 corresponding to a position of the first mounting groove 141 and a sixth mating structure 126 corresponding to a position of the second mounting groove 142 are disposed on a surface of the other of the first and second

sub enclosing plates

121 and 122 facing the receiving groove 140, a seventh mating structure (not shown in the drawings) rotatably connected to the fifth mating structure 125 is disposed on the first latch shaft 312 adjacent to the fifth mating structure 125, and an eighth mating structure (not shown in the drawings) rotatably connected to the sixth mating structure 126 is disposed on the second latch shaft 322 adjacent to the sixth mating structure 126. In an example, the fifth mating structure 125 may be one of a male shaft and a female receptacle, the seventh mating structure may be the other of a male shaft or a female receptacle, the sixth mating structure 126 may be one of a male shaft and a female receptacle, and the eighth mating structure may be the other of a male shaft or a female receptacle.

In some exemplary embodiments, the second and seventh mating structures 313 and the seventh mating structures have the same structure, and may be both cross slots, straight latches, triangular latches, hexagonal latches, and the like, and the fourth and eighth mating structures have the same structure, and may be both cross slots, straight latches, triangular latches, hexagonal latches, and the like.

In some exemplary embodiments, as shown in fig. 2, the first roller 310 is provided with a fifth locking groove 314, the second roller 320 is provided with a sixth locking groove (not shown in the drawings), the first end 201 of the elastic base film 200 is provided with a first clip 210 that mates with the fifth locking groove 314, and the second end 202 of the elastic base film 200 is provided with a second clip 220 that mates with the sixth locking groove. The fifth locking grooves 314 may be plural and spaced apart in the second direction, the sixth locking grooves may be plural and spaced apart in the second direction, and the number of the fifth locking grooves 314 and the sixth locking grooves is the same as that of the first barrier 131. In this example, the second direction may be an extending direction of the first and

second rollers

310 and 320. The first clip 210 and the fifth clip groove 314 are detachably connected, and the second clip 220 and the sixth clip groove are detachably connected, so that the elastic basement membrane 200 can be conveniently replaced and removed for other experiments. The first clip 210 and the second clip 220 may be both a linear clip, the fifth card slot 314 is adapted to the first clip 210, the sixth card slot is adapted to the second clip 220, and 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 film 200.

Fig. 8 is a cross-sectional view taken at the position a-a in fig. 7. In some exemplary embodiments, as shown in fig. 7 and 8, a notch position of the fifth card slot 314 is provided with a first limiting groove 315, a notch position of the sixth card slot is provided with a second limiting groove, a width of the fifth card slot 314 is greater than a width of the first limiting groove 315 in the second direction, the width of the first limiting groove 315 is adapted to the width of the elastic base film 200 and is configured to limit the movement of the elastic base film 200 in the second direction, the width of the sixth card slot is greater than the width of the second limiting groove, and the width of the second limiting groove is adapted to the width of the elastic base film 200 and is configured to limit the movement of the elastic base film 200 in the second direction.

In some exemplary embodiments, as shown in fig. 2, the cell culture apparatus 1 further includes a cover plate 400, and the cover plate 400 covers the outer side of the enclosing plate 120 and covers the receiving groove 140. The cover plate 400 includes a top plate 410 and a side plate 420 disposed on a circumference of the top plate 410, and the side plate 420 is located on a side of the enclosing plate 120 away from the receiving groove 140. The cover plate 400 may be made of a transparent material to facilitate observation of the state of cells in the culture chamber.

In some exemplary embodiments, as shown in fig. 2, the periphery of the bottom plate 110 protrudes out of the enclosing plate 120 along a direction away from the receiving groove 140, and the side plate 420 abuts against the bottom plate 110.

In some exemplary embodiments, as shown in fig. 2, the side plate 420 is provided with a first escape groove 421 for escaping the first driving shaft 330 and a second escape groove 422 for escaping the second driving shaft 340.

In some exemplary embodiments, as shown in FIG. 2, 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 fluid exchange port 430 includes an inlet port 431 and an outlet port 432. The liquid inlet 431 can be communicated with the outlet of the liquid supply pump through a liquid supply pipeline, and the liquid outlet 432 can be communicated with the inlet of the liquid discharge pump through a liquid discharge pipeline, so that a small cell workstation can be expanded, and simple cell liquid changing, drug screening and other works can be completed. In order to ensure a sterile environment in the culture chamber 150, the liquid exchange port 430 may be provided with a sealing ring that seals a gap between the top plate 410 and the pipes (the drain pipe and the supply pipe).

In some exemplary embodiments, as shown in FIGS. 2 and 3a, a first anti-reflection structure 127 is provided on the shroud 120 and a second anti-reflection structure 440 is provided on the cover 400 that cooperates with the first anti-reflection structure 127. The first and second

anti-reflection structures

127 and 440 can prevent the cover plate 400 and the culture dish 100 from being reversely mounted, so that the first and second avoiding

grooves

421 and 340 do not correspond to the first driving shaft 330 and the second and second avoiding

grooves

422 and 340, respectively, and the first and

second driving shafts

330 and 340 are damaged. The first anti-wind back structure 127 may be a first wedge angle disposed on the first shroud 120 and the second anti-wind back structure 440 may be a second wedge angle disposed on the side plate 420.

In some exemplary embodiments, as shown in FIG. 3a, the distance between the surface of the partition 130 remote from the base plate 110 and the base plate 110 is greater than the distance between the surface of the enclosure 120 remote from the base plate 110 and the base plate 110. In an example, the cell culture apparatus 1 further includes a first convex edge 1231 disposed on the third sub-enclosing plate 123 and a second convex edge 1241 disposed on the fourth sub-enclosing plate 124, the first convex edge 1231 and the second convex edge 1241 are disposed along the edge of the accommodating groove 140, and a distance between a surface of the first convex edge 1231 and the second convex edge 1241 away from the bottom plate 110 and the bottom plate 110 is equal to a distance between a surface of the partition plate 130 away from the bottom plate 110 and the bottom plate 110.

The following describes the technical means of the cell culture apparatus 1 according to the embodiment of the present invention by exemplifying the operation principle of the cell culture apparatus 1.

In some exemplary embodiments, the elastic base membrane 200 corresponds to the number and position of the first partition plates 131, the elastic base membrane 200 is inserted into the corresponding first through hole 161 through the corresponding first through channel 162, the elastic base membrane 200 is inserted into the corresponding second through hole through the corresponding second through channel 163, the elastic base membrane 200 covers the bottom of the culture chamber 150, the first clip 210 at the first end 201 of the elastic base membrane 200 is inserted into the fifth slot 314 corresponding to the first roller 310, and the second clip 220 at the second end 202 of the elastic base membrane 200 is inserted into the sixth slot corresponding to the second roller 320, so as to fix the elastic base membrane 200. Then, the first shaft 312 of the first roller 310 is engaged into the first engaging portion 1772 of the first engaging seat 171 through the first guiding portion 1771 and is retained by the first engaging portion 1772, and the second shaft 322 of the second roller 320 is engaged into the second engaging portion 1782 of the second engaging seat 172 through the second guiding portion 1781 and is retained by the second engaging portion 1782. The first engaging structure 331 of the first driving shaft 330 is engaged with the second engaging structure 313 of the first engaging shaft 312 adjacent to the first driving shaft 330, the fifth engaging structure 125 is rotatably connected with the seventh engaging structure of the first engaging shaft 312 away from the first driving shaft 330, the third engaging portion of the second driving shaft 340 is engaged with the fourth engaging portion of the second engaging shaft 322 adjacent to the second driving shaft 340, the sixth engaging structure 126 is rotatably connected with the eighth engaging structure of the second engaging shaft 322 away from the second driving shaft 340, and the first driving shaft 330 can actuate the first roller 310, the second driving shaft 340 can actuate the second roller 320, and the elastic base film 200 can be stretched or relaxed when the first driving shaft 330 and the second driving shaft 340 rotate in opposite directions. In 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-enclosing plate 121 and the third sub-enclosing plate 122 may be pulled outwards when the first roller 310 and the second roller 320 are installed, so that the first sub-enclosing plate 121 and the third sub-enclosing plate 122 are elastically deformed to expand the space of the first installation groove 141 and the second installation groove 142. The cover plate 400 is covered, the first avoiding groove 421 of the cover plate 400 corresponds to the first driving shaft 330, and the second avoiding groove 422 of the cover plate 400 corresponds to the second driving shaft 340. Finally, a liquid supply pipe is inserted into the liquid inlet 431 of the cover plate 400, a liquid discharge pipe is inserted into the liquid outlet 432 of the cover plate 400, and the other ends of the liquid supply pipe and the liquid discharge pipe are connected with corresponding pumps.

An amount of a cell adhesion promoting substance, such as Polylysine (PLL) or collagen, is pumped into the culture chamber 150. Pumping a certain amount of Phosphate Buffered Saline (PBS) solution to clean residual liquid, pumping cell suspension with a certain cell density, standing for more than 6 hours until the cells are attached to the elastic basement membrane 200, and then performing conventional cell culture.

Finally, according to the experimental requirements, in combination with hooke's law, the first roller 310 is driven to rotate by the first actuator 332, the second roller 320 is driven to rotate by the second actuator 342, so as to stretch the elastic basement membrane 200 by a predetermined length, thereby achieving the stretching effect on the cells to simulate the growth environment of the cells in the body, for example, as shown in fig. 9a and 9b, the first actuator 332 drives the first roller 310 to rotate along the clockwise direction, the second actuator 342 drives the second roller 320 to rotate along the counterclockwise direction, the elastic basement membrane 200 is stretched, the cells attached to the elastic basement membrane 200 are stretched, then the first actuator 332 drives the first roller 310 to rotate along the counterclockwise direction, when the second actuator 342 drives the second roller 320 to rotate clockwise, the elastic basement membrane 200 relaxes from the stretched state to the original state, the cells attached to the elastic basement membrane 200 also return to the static state, the above actions are repeated to make the cells in a dynamic environment.

It can be seen from the operation principle of the cell culture apparatus 1 according to the exemplary embodiment of the 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, and the elastic basement membrane 200 can stretch or relax during the rotation of the first roller 310 and the second roller 320, so that the cells attached to the elastic basement membrane 200 can synchronously move with the elastic basement membrane 200, and the cells can be cultured in a dynamic environment, thereby truly reflecting the state of the cells cultured in vivo.

Although the embodiments of the present invention have been described above, the above description is only for the convenience of understanding the present invention, and is not intended to limit the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. A cell culture apparatus, comprising:

the culture dish comprises a bottom plate, a surrounding plate and at least one partition plate, wherein the surrounding plate is arranged along the circumferential direction of the bottom plate, the bottom plate and the surrounding plate form a containing groove, the partition plate is arranged in the containing groove and limits at least one culture chamber, and a penetrating hole communicated with the culture chamber is formed between the partition plate and the bottom plate;

the elastic basement membrane penetrates through the penetrating hole and covers the position, corresponding to the culture chamber, of the bottom plate;

and the actuating mechanism is at least partially arranged in the accommodating groove, is positioned between the enclosing plate and the partition plate and is used for stretching the elastic base film.

2. The cell culture apparatus of claim 1, wherein: the partition board comprises a first partition board, the first partition board comprises a first sub-partition board extending along a first direction and a second sub-partition board and a third sub-partition board, the first sub-partition board is arranged on the first sub-partition board at intervals along the first direction, the second sub-partition board and the bottom board are arranged at intervals, intervals are arranged between the second sub-partition board and the bottom board, intervals are arranged between the third sub-partition board and the bottom board, the through holes comprise first through holes between the second sub-partition board and the bottom board and second through holes between the third sub-partition board and the bottom board, and the first direction and the second direction are intersected.

3. The cell culture apparatus of claim 2, wherein: in the direction perpendicular to the bottom plate, the aperture of the through hole is matched with the thickness of the elastic basement membrane, and in the second direction, the aperture of the through hole is matched with the width of the elastic basement membrane.

4. The cell culture apparatus of claim 2, wherein: the first partition plate includes a plurality of partition plates arranged at intervals in the second direction.

5. The cell culture apparatus according to any one of claims 2 to 4, wherein: the coaming includes the edge first direction extends and follows first sub-coaming and the second sub-coaming that the second direction interval set up and follow the second direction extends and follows third sub-coaming and the fourth sub-coaming that the first direction interval set up, the third sub-coaming with the second sub-baffle is adjacent, the fourth sub-coaming with the third sub-baffle is adjacent, the third sub-coaming with form between the second sub-baffle with the first mounting groove of first through-hole intercommunication, the fourth sub-coaming with form between the third sub-baffle with the second mounting groove of second through-hole intercommunication.

6. The cell culture apparatus of claim 5, wherein: actuating mechanism including install in first roller bearing in the first mounting groove with install in the second roller bearing in the second mounting groove, the elasticity basement membrane includes along first direction interval set up first end and second end, first end with first roller bearing is connected, the second end with the second roller bearing is connected.

7. The cell culture apparatus of claim 6, wherein: the first roller comprises a first shaft body and first clamping shafts arranged at two ends of the first shaft body, two first clamping seats are arranged in the first mounting groove, the first shaft body is arranged between the first clamping seats, the first clamping shafts are arranged in the corresponding first clamping seats, the second roller comprises a second shaft body and second clamping shafts arranged at two ends of the second shaft body, two second clamping seats are arranged in the second mounting groove, the second shaft body is arranged between the second clamping seats, and the second clamping shafts are arranged in the corresponding second clamping seats.

8. The cell culture apparatus of claim 7, wherein: the first sub-baffle plate of the first baffle plate adjacent to the first sub-enclosing plate extends into the first mounting groove to form a first extending portion, and extends into the second mounting groove to form a second extending portion, one of the two first clamping seats comprises the first extending portion and a first clamping groove arranged on the end face, far away from the bottom plate, of the first extending portion, one of the two second clamping seats comprises the second extending portion and a second clamping groove arranged on the end face, far away from the bottom plate, of the second extending portion, the first clamping shaft is embedded into the first clamping groove and can rotate, and the second clamping shaft is embedded into the second clamping groove and can rotate.

9. The cell culture apparatus of claim 8, wherein: the baffle is characterized by further comprising a second baffle arranged between the first baffle and the second sub-enclosing plate, the second baffle comprises a third extending part extending into the first mounting groove and a fourth extending part extending into the second mounting groove, the other of the first clamping seats comprises the third extending part and a third clamping groove arranged on the end face of the bottom plate far away from the third extending part, the other of the second clamping seats comprises the fourth extending part and a fourth clamping groove arranged on the end face of the bottom plate far away from the fourth extending part, the first clamping shaft is embedded into the third clamping groove and can rotate, and the second clamping shaft is embedded into the fourth clamping groove and can rotate.

10. The cell culture apparatus of claim 7, wherein: a first shaft hole communicated with the first mounting groove and a second shaft hole communicated with the second mounting groove are formed in the surface, far away from the accommodating groove, of one of the first sub surrounding plate and the second sub surrounding plate, the actuating mechanism comprises a first driving shaft penetrating through the first shaft hole and a second driving shaft penetrating through the second shaft hole, a first matching structure is arranged at one end, close to the first clamping shaft, of the first driving shaft, a second matching structure clamped with the first matching structure is arranged on the first clamping shaft, close to the first driving shaft, of the first driving shaft, the first driving shaft is arranged to be capable of actuating the first rolling shaft after the first matching structure and the second matching structure are clamped, a third matching structure is arranged at one end, close to the second clamping shaft, of the second driving shaft, and a fourth matching structure clamped with the third matching structure is arranged on the second clamping shaft, close to the second driving shaft, the second driving shaft is used for actuating the second roller after the third matching structure and the fourth matching structure are clamped.

11. The cell culture apparatus of claim 10, wherein: the first driving shaft is provided with a first driving piece at one end far away from the first roller, the second driving shaft is provided with a second driving piece at one end far away from the second roller, the first driving piece is used for driving the first driving shaft to drive the first roller to rotate, and the second driving piece is used for driving the second driving shaft to drive the second roller to rotate.

12. The cell culture apparatus of claim 10, wherein: the surface of the other one of the first sub enclosing plate and the second sub enclosing plate facing the accommodating groove is provided with a fifth matching structure corresponding to the first mounting groove and a sixth matching structure corresponding to the second mounting groove, a seventh matching structure rotatably connected with the fifth matching structure is arranged on a first clamping shaft adjacent to the fifth matching structure, and an eighth matching structure rotatably connected with the sixth matching structure is arranged on a second clamping shaft adjacent to the sixth matching structure.

13. The cell culture apparatus of claim 6, wherein: the first roller bearing is provided with a fifth clamping groove, the second roller bearing is provided with a sixth clamping groove, the first end of the elastic basement membrane is provided with a first clamping matched with the fifth clamping groove, and the second end of the elastic basement membrane is provided with a second clamping matched with the sixth clamping groove.

14. The cell culture apparatus of claim 13, wherein: the notch position of fifth draw-in groove is provided with first spacing groove, the notch position of sixth draw-in groove is provided with the second spacing groove on the second direction, the width of fifth draw-in groove is greater than the width of first spacing groove, the width of first spacing groove with the width adaptation of elasticity basement membrane to set up to the restriction the removal of elasticity basement membrane in the second direction, the width of sixth draw-in groove is greater than the width of second spacing groove, the width of second spacing groove with the width adaptation of elasticity basement membrane, and set up to the restriction the elasticity basement membrane is in the ascending removal of second direction.

15. The cell culture apparatus of claim 1, wherein: the cell culture device still includes the apron, the apron cover is located the outside of bounding wall covers the storage tank, the apron include the roof with set up in roof ascending curb plate in week, the curb plate is located the bounding wall is kept away from one side of storage tank, be provided with the liquid exchange mouth on the roof, the liquid exchange mouth with cultivate the room and correspond.