Automated cell culture system and automated cell culture method based on same

Abstract

A method for culturing cells through a cell culture supply device according to an embodiment of the present application comprises the steps of: seeding cells into a cartridge; coupling the cartridge to a platform; injecting a culture medium into the cartridge coupled to the platform; and circulating the culture medium in the platform, wherein the step of seeding cells in the cartridge comprises the steps of: coupling a middle layer included in the cartridge to a seeding kit; and seeding cells into different areas in the middle layer.

Description

Cell culture automation system and cell culture automation method based thereon

[0001] The present invention relates to a culture solution supply device capable of supplying a culture solution to a cell culture chip, and more specifically, to an automated cell culture process based on the culture solution supply device.

[0002]

[0003] Cell culture systems play an essential role in life science research and new drug development, artificially culturing cells to enable a variety of biological experiments. These systems encompass a series of processes, including cell seeding, cell culture, cell analysis, and monitoring, and can help cells grow and differentiate under conditions similar to the biological environment. While traditional cell culture methods require researchers to manually perform multiple steps, systems are being developed that automate these processes, significantly improving efficiency and accuracy.

[0004] Automation technology in cell culture systems plays a crucial role, helping researchers efficiently conduct large-scale experiments while maintaining consistent experimental conditions. In particular, automated processes can reduce research time and costs and significantly improve the reliability of experimental data.

[0005] However, conventional cell culture systems rely on manual labor by researchers, making them prone to human error during the experiment. This can lead to uneven cell culture, inconsistent experimental results, and reduced reproducibility. To address these issues, the development of technologies that automate the entire cell seeding, culture, analysis, and monitoring process is essential. In particular, it is necessary to establish product usage processes that minimize human error, thereby enhancing experimental accuracy and efficiency.

[0006]

[0007] One object of the present invention is to provide a culture medium supply device capable of actively controlling the culture medium supply speed and simultaneously supplying different culture mediums to multiple chips at an accurate speed and in an accurate amount for each of two types of cells co-cultured in a cell culture chip.

[0008] One object of the present invention is to provide a device capable of implementing physiological conditions identical to those of human organs by supplying two types of culture media at an accurate flow rate without a tube, and capable of simultaneously supplying culture media to multiple chips in one layer and multiple chips in multiple layers using a single actuator.

[0009] One object of the present invention is to provide a cell culture system and to provide an automated cell culture process based on the cell culture system.

[0010] The problems to be solved by the present invention are not limited to the problems described above, and problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention pertains from this specification and the attached drawings.

[0011]

[0012] According to one embodiment, a culture medium supply device for a cell culture chip includes a base having a rotatably driven ring-shaped ring gear, a top cover provided on an upper portion of the base, on which a cell culture chip is mounted at an upper edge, a culture medium reservoir provided on an upper surface, and a plurality of channels formed on a rear surface, and a membrane attached to a rear surface of the top cover, on which a channel-shaped driving pressure transmission unit is formed, and the ring gear includes a plurality of driving pressure units that protrude upward and pressurize and deform the driving pressure transmission unit so that the driving pressure transmission unit performs a pumping operation.

[0013] According to one embodiment, a method of culturing cells using a cell culture supply device comprises the steps of seeding cells in a cartridge, coupling the cartridge to a platform, injecting a culture medium into the cartridge coupled to the platform, and circulating the culture medium within the platform, wherein the step of seeding cells in the cartridge comprises the steps of coupling a middle layer included in the cartridge to a seeding kit, and seeding cells in different regions of the middle layer.

[0014] The solutions to the problems of the present invention are not limited to the solutions described above, and solutions that are not mentioned can be clearly understood by a person having ordinary skill in the art to which the present invention pertains from this specification and the attached drawings.

[0015]

[0016] According to one embodiment, a culture medium is supplied to a cell culture chip, and different culture mediums can be supplied simultaneously to two or more cells in the cell culture chip by means of a ring gear of a cell solution supply device and two or more driving transmission units of a membrane that is pressurized by the ring gear and performs a pumping operation.

[0017] According to one embodiment, in order to supply two or more culture media at different rates within a cell culture chip, the internal cross-sectional areas of two driving transmission units are formed differently, thereby supplying culture media to two or more cells within the cell culture chip at different rates.

[0018] According to one embodiment, a cell culture chip, a chip holder, and a new cell culture device are connected without requiring separate tubing work, thereby solving the problem of the supply amount of culture medium differing from the setting according to the tubing length, and initial setting can be made easy.

[0019] In one embodiment, since the culture solution is supplied in a circulating structure, it is possible to implement the structure and function of the human body, thereby enabling in-vitro physiological and pathological simulations similar to the human body, thereby increasing the accuracy of drug testing.

[0020] According to one embodiment, the tubingless structure prevents bubbles from forming in the chip, reduces dead volume to reduce the amount of culture medium used, enables large-scale processing with a structure that allows an increase in the number of chips, and prevents contamination of the culture medium in the chip by supplying it without direct contact with the culture medium.

[0021] In one embodiment, by providing an automated process, the accuracy and efficiency of the experiment can be greatly improved, and the consistency and reproducibility of the experimental results can be increased by minimizing human error.

[0022] The effects of the present invention are not limited to the effects described above, and effects not mentioned can be clearly understood by a person having ordinary skill in the art to which the present invention pertains from this specification and the attached drawings.

[0023]

[0024] Figure 1 is a drawing for explaining the cell culture system of the present invention.

[0025] Fig. 2A is a drawing for explaining the combined structure of the cell culture chip, chip holder, and channel plate shown in Fig. 1, and Fig. 2B is a drawing for explaining the completed combined form in Fig. 2A.

[0026] Figure 3 is a drawing for explaining the cell culture chip shown in Figures 2A and 2B.

[0027] Figure 4 is a drawing for explaining an example of co-culture in which two different cells are placed in the cell culture unit illustrated in Figure 3.

[0028] FIG. 5A is an exploded perspective view showing another embodiment of the channel plate in FIGS. 2A and 2B, and FIG. 5B is a drawing showing the combined state of FIG. 5A.

[0029] Figure 6 is a perspective view of the culture solution supply device illustrated in Figure 1.

[0030] Fig. 7 is a drawing for explaining the gear part shown in Fig. 6.

[0031] Fig. 8 is a drawing for explaining the state in which the gear part shown in Fig. 7 is mounted.

[0032] Figure 9A is a drawing for explaining the upper surface of the cell culture system shown in Figure 1, and Figure 9B is a drawing for showing the state in which the chip holder cover of the upper part of the cell culture system is open.

[0033] Figure 10 is a drawing for explaining the channel shown in Figure 9B.

[0034] Figure 11 is a drawing for explaining a driving pressure transmission unit formed on a channel plate.

[0035] Fig. 12 is a cross-sectional view of the driving pressure transmission unit illustrated in Fig. 11.

[0036] Figure 13 is a drawing for explaining the joint structure of a channel plate and an elastomer channel.

[0037] Figures 14A to 14C are drawings for explaining the pumping operation for supplying and recovering culture medium.

[0038] Figure 15 is a drawing for explaining a culture solution supply device for a cell culture chip having a structure in which culture solution supply units are stacked.

[0039] Fig. 16 is a drawing showing the appearance of a culture solution supply device and a chip holder mounting portion for a cell culture chip according to Fig. 15.

[0040] Figures 17A and 17B are drawings for explaining the operation of the chip holder mounting part.

[0041] Fig. 18 is a drawing for explaining another embodiment of a chip holder mounting part.

[0042] Figures 19A to 19F are drawings for explaining the process of mounting a channel plate on a culture solution supply device for a cell culture chip according to Figure 16.

[0043] FIG. 20 is a drawing for explaining a cell culture system, analysis device, and user terminal configuration according to one embodiment.

[0044] Figures 21 to 24 are drawings for explaining the configuration of a cell culture system according to one embodiment.

[0045] FIG. 25 is a drawing for explaining a method of culturing cells using a cell culture supply device according to one embodiment.

[0046] FIG. 26 is a drawing for detailing the cell seeding and stabilization steps according to one embodiment.

[0047] Figure 27 is a drawing for explaining a loading step according to one embodiment.

[0048] FIG. 28 is a drawing illustrating a step of circulating a culture medium in a cartridge within a platform according to one embodiment.

[0049] FIG. 29 is a drawing for explaining a step of replacing and sampling a culture medium according to one embodiment.

[0050] FIG. 30 is a diagram illustrating steps for performing analysis on cells according to one embodiment.

[0051]

[0052] The above-described purposes, features, and advantages of the present application will become more apparent through the following detailed description, taken in conjunction with the accompanying drawings. However, as the present application is susceptible to various modifications and various embodiments, specific embodiments will be illustrated in the drawings and described in detail below.

[0053] Throughout the specification, identical reference numerals, in principle, represent identical components. Furthermore, components with identical functions within the scope of the same concept shown in the drawings of each embodiment are described using the same reference numerals, and redundant descriptions thereof will be omitted.

[0054] If a detailed description of a known function or configuration related to this application is deemed to unnecessarily obscure the gist of this application, such detailed description will be omitted. Furthermore, numbers (e.g., "first," "second," etc.) used throughout the description of this specification are merely identifiers used to distinguish one component from another.

[0055] In addition, the suffixes "module" and "part" for components used in the following examples are given or used interchangeably only for the convenience of writing the specification, and do not have distinct meanings or roles in themselves.

[0056] In the examples below, singular expressions include plural expressions unless the context clearly indicates otherwise.

[0057] In the examples below, terms such as “include” or “have” mean that a feature or component described in the specification is present, and do not preclude the possibility that one or more other features or components may be added.

[0058] For convenience of explanation, the sizes of components in the drawings may be exaggerated or reduced. For example, the sizes and thicknesses of each component shown in the drawings are arbitrarily shown for convenience of explanation, and the present invention is not necessarily limited to what is shown.

[0059] In some embodiments, where implementations are otherwise feasible, the order of specific processes may differ from the order described. For example, two processes described in succession may be performed substantially simultaneously, or in a reverse order from the order described.

[0060] In the following examples, when components are said to be connected, this includes not only cases where the components are directly connected, but also cases where components are interposed between the components and are indirectly connected.

[0061] For example, when it is said in this specification that components, etc. are electrically connected, it includes not only cases where the components, etc. are directly electrically connected, but also cases where components, etc. are interposed and indirectly electrically connected.

[0062]

[0063] FIG. 1 is a drawing for explaining a cell culture device of the present invention, and FIG. 2 is a drawing for explaining a cell culture chip and chip holder illustrated in FIG. 1.

[0064] Referring to FIG. 1, FIG. 2A and FIG. 2B, a cell culture device (10) includes a cell culture chip (100), a chip holder (200) in which the cell culture chip (100) is mounted, a channel plate (500) in which the chip holder (200) is mounted, and a culture solution supply device (300) in which the channel plate (500) is accommodated.

[0065] The cell culture device (10) is connected to a computer or mobile device and can be remotely controlled by the computer or mobile device.

[0066] According to the present invention, a culture medium is supplied to a cell culture chip (100), and different culture mediums can be supplied simultaneously to two or more cells in the cell culture chip (100) by means of a ring gear (333) of a culture medium supply device (300) and a drive transmission unit (510) of a channel plate (500) that is pressurized by the ring gear (333) and performs a pumping operation.

[0067] In addition, according to the present invention, in order to supply two or more culture media at different speeds within the cell culture chip (100), the internal cross-sectional areas of the two driving transmission units (510) are formed differently, thereby supplying the culture media to two or more cells within the cell culture chip (100) at different speeds.

[0068] In addition, according to the present invention, a cell culture chip (100), a chip holder (200), and a new cell culture device (300) are connected without requiring separate tubing work, thereby solving the problem of the supply amount of culture solution differing from the setting according to the tubing length, and has the advantage of simple initial setting.

[0069] Below, the detailed configuration of the cell culture device of the present invention is described with reference to the drawings.

[0070]

[0071] [Cell culture chip]

[0072] FIG. 3 is a drawing for explaining the cell culture chip illustrated in FIG. 2A and FIG. 2B, and FIG. 4 is a drawing for explaining an example of co-culture in which two different cells are placed in the cell culture unit (121) illustrated in FIG. 3.

[0073] First, referring to FIG. 3, the cell culture chip (100) includes a structure in which a lower layer (130), a middle layer (120), and an upper layer (110) are sequentially stacked.

[0074] The middle layer (120) includes a cell culture section (121) composed of a membrane in the middle portion on a horizontal plane, and different types of cells can be co-cultured on the upper and lower surfaces of the cell culture section (121).

[0075] For example, in the case of co-culture as shown in FIG. 4, epithelial cells (Epithelium cells) as Cell Type 1 may be attached to the upper surface of the middle layer (120), and endothelial cells (Endothelium) as Cell Type 2 may be attached to the back surface. In addition, an extracellular matrix (ECM) may be placed between the epithelial cells and the upper surface of the cell culture unit (121), and between the endothelial cells and the back surface of the cell culture unit (121).

[0076] Referring again to FIG. 3, an upper channel (140) is formed in the upper layer (110) as a path for supplying and recovering culture medium to the first type cells placed on the upper surface of the cell culture unit (121).

[0077] At both ends of the upper channel (140), a first supply hole (115) through which the first type culture medium is supplied and a first discharge hole (116) through which the first type culture medium is discharged are formed.

[0078] In the lower layer (130), a lower channel (150) is formed as a path for supplying and recovering culture medium to the second type cells placed on the back surface of the cell culture unit (121).

[0079] At both ends of the lower channel (150), a second supply hole (135) for supplying the second type culture medium and a second discharge hole (136) for discharging the second type culture medium are formed.

[0080] The first supply hole (115), the first discharge hole (116), the second supply hole (135), and the second discharge hole (136) are respectively connected to the first supply connection (531), the first discharge connection (532), the second supply connection (533), and the second discharge connection (534) of the channel plate (500).

[0081]

[0082] [Chip Holder]

[0083] Referring to FIGS. 2A and 2B, the chip holder (200) fixes the cell culture chip (100) and is provided so that it can be mounted on a channel plate (500).

[0084] The chip holder (200) includes an upper member (210) and a lower member (220), and the upper member (210) and the lower member (220) are fixedly joined to each other with the cell culture chip (100) interposed therebetween.

[0085] The upper substrate (210) and the lower substrate (220) can be fixed and combined by applying a sliding method. At this time, the individual layers of the cell culture chip (100) are not adhered to each other, but are merely fixed by the combination of the upper substrate (210) and the lower substrate (220) of the chip holder (200) in a combined state, and then mounted on the channel plate (500).

[0086] The chip holder (200) is configured so that the first supply hole (115), the first discharge hole (116), the second supply hole (135), and the second discharge hole (136) formed in the cell culture chip (100) through an internal path can be connected to the first supply connection part (531), the first discharge connection part (532), the second supply connection part (533), and the second discharge connection part (534) of the channel plate (500), respectively.

[0087] In particular, since a leak prevention member such as a rubber O-ring is provided in each of the first supply connection part (531), the first discharge connection part (532), the second supply connection part (533), and the second discharge connection part (534), leakage of the culture solution and pressure loss can be prevented during circulation of the culture solution. The reservoir (540) holds the culture solution required for each chip, and the supply and discharge are performed in the same reservoir so that a circulating culture solution supply method is possible. A culture solution storage set (540) composed of a first culture solution storage part (541) in which the first culture solution is stored and a second culture solution storage part (542) in which the second culture solution is stored as a pair is provided in each chip holder.

[0088] According to the present invention, the first supply connection part (531), the first discharge connection part (532), the second supply connection part (533), and the second discharge connection part (534) of the channel plate (500) are each connected to the culture medium reservoir (540) and the pipes are connected to each other, so that separate tubing work is not required, and pressure loss due to separate tubing does not occur, enabling precise supply control of the culture medium. In addition, the flow of the culture medium is circulated in a closed loop within the chip and holder, so that contamination from the outside can be prevented.

[0089] Meanwhile, referring to FIGS. 5A and 5B, a cell culture chip (100) can be accommodated in a channel plate (500) without a chip holder.

[0090] That is, according to the embodiment shown in FIGS. 5A and 5B, the channel plate may be composed of an upper channel plate (570) and a lower channel plate (580), and the culture medium stored in the reservoir (540) may be moved to the second supply hole (584) where the cell culture chip is located through the first supply hole (582), and the pumped culture medium may be discharged through the discharge hole (572) and re-enter the reservoir (540).

[0091]

[0092] [Culture solution supply device]

[0093] Figure 6 is a perspective view of the culture solution supply device illustrated in Figure 1.

[0094] Referring to FIG. 6, the culture solution supply device (300) includes a base portion (310), a top cover (360), and a gear portion (330).

[0095] The base part (310) includes a gear mounting part (320), a rotary driving part (312), and a base cover part (315).

[0096] An internal space (311) capable of accommodating a gear mounting portion (320) is provided inside the above base cover portion (315), a rotary drive portion (312) is installed in the center of the internal space, and a gear mounting portion (320) is provided on the upper portion of the rotary drive portion (312).

[0097] The rotary drive unit (312) is a means for generating rotary driving force to transmit driving force, and may be provided with any one of various types of motors, such as an AC motor, DC motor, or stepper motor, having a rotary driving shaft.

[0098] The gear mounting portion (320) has a motor shaft through hole (321) through which the drive shaft of the rotary drive portion (312) passes and protrudes in the center, and a gear mounting groove (322) formed in a groove shape corresponding to the gear portion (330) and in which the gear portion (330) is mounted.

[0099] FIG. 7 is a drawing for explaining the gear part (330) illustrated in FIG. 6, and FIG. 8 is a drawing for explaining the state in which the gear part illustrated in FIG. 7 is mounted.

[0100] Referring to FIGS. 6 to 8, the gear unit (330) includes a sun gear (331) arranged in the center and connected to a rotary drive shaft through a keyway type coupling, a plurality of planetary gears (332) arranged on the outside of the sun gear (331) and rotatably driven by meshing with the sun gear (331), and a ring gear (333) in the shape of a ring that rotatably drives by meshing with all of the plurality of planetary gears (332). The sun gear (331) and the planetary gears (332) rotate in different rotational directions, while the planetary gears (332) and the ring gear (333) rotate in the same rotational direction. The sun gear (331) exhibits a rotational motion in which the position does not change, but the planetary gears (332) exhibit a rotational motion in which the position changes.

[0101] Referring to Fig. 7, a ring body (336) that moves together with the ring gear is coupled to the outer circumference of the ring gear (333), and a plurality of driving pressure parts (335) that protrude upward along the circumference are formed on the ring body (336).

[0102] The driving pressure unit (335) pressurizes the driving pressure transmission unit (510) formed on the bottom surface of the channel plate (500), and the area pressurized by the driving pressure transmission unit (510) changes depending on the movement of the driving pressure unit (335). Details regarding this will be described later in relation to the pumping operation for supplying and recovering the culture medium. The driving pressure unit (335) may also be formed on the upper portion of the ring gear without a separate ring body.

[0103] Meanwhile, in the case where the sun gear (331), multiple planetary gears (332), and ring gear (333) are interlocked and rotate, a uniform pressure can be applied to each of the multiple driving pressure transmission parts (510) of the channel plate (500) compared to when multiple driving pressures (335) are provided on the edge of one disk connected to the rotational drive shaft.

[0104] The gear cover (340) performs the function of protecting the gear portion (330). The gear cover (340) covers the portion except for the plurality of driving pressure portions (335) that protrude upward along the edge of the ring gear (333). Accordingly, the plurality of driving pressure portions (335) protrude upward from the outer portion of the gear cover (340).

[0105] Here, the gear mounting groove (322) in which the gear portion (330) is mounted is composed of a plurality of grooves formed according to the position and size corresponding to each gear, and the portion where the gear is not placed protrudes upward to support the gear cover (340) and prevent the gear cover (340) from rotating.

[0106] Figures 9A and 9B are drawings for explaining the top cover (360), which is the upper surface of a cell culture device equipped with a channel plate (500).

[0107] The upper edge of the above cell culture device is provided with a chip holder cover (364) that can be opened and closed. When the channel plate is mounted, the channel plate (500) containing the chip holder can be inserted from above or from the side while the chip holder cover (364) is open. When the insertion of the channel plate is completed, the chip holder cover (364) closes the upper part of the chip holder and a culture medium can be supplied.

[0108] According to the present invention, since the chip holder (200) is detachably connected to the culture solution supply device (300) and the pipe connection for supplying / retrieving the culture solution is completed, no separate tubing work is required and no pressure loss occurs due to the separate pipe connection, enabling precise supply control of the culture solution.

[0109] Referring to Fig. 10, a plurality of channels (367), holes (318), and driving pressure transmission holes (370) that serve as passages for supplying and recovering culture medium are formed on the back surface of the channel plate (500).

[0110] FIG. 11 is a drawing for explaining a membrane (580) attached to the back surface of the channel plate (500) shown in FIG. 10, and a driving pressure transmission unit (510) is formed on the membrane. In addition, FIG. 12 is a cross-sectional view of the driving pressure transmission unit (510) shown in FIG. 11.

[0111] Referring to FIG. 11, the driving pressure transmission part (510) attached to the driving pressure transmission hole (370) of the channel plate (500) forms a channel, protrudes toward the bottom of the membrane, and may be made of a material such as an elastomer that is flexible and can change shape.

[0112] The driving pressure transmission unit (510) includes first and second driving pressure transmission units (510a, 510b), and the channel plate (500) may include either of the first and second driving pressure transmission units (510a, 510b).

[0113] In addition, the internal cross-sectional areas of the first and second driving pressure transmission units (510a, 510b) can be formed differently, so that the supply speeds of the first culture medium and the second culture medium can be made different.

[0114] For example, by setting the internal cross-sectional area of ​​the first driving pressure transmission unit (510a) to be larger than that of the second driving pressure transmission unit (510b), the supply speed of the first culture medium can be made faster than that of the second culture medium during one pumping cycle. Or, conversely, by setting the internal cross-sectional area of ​​the first driving pressure transmission unit (510a) to be smaller than that of the second driving pressure transmission unit (510b), the supply speed of the first culture medium can be made slower than that of the second culture medium during one pumping cycle.

[0115] In addition, the first and second culture medium supply speeds can be made different by forming the positions of the first and second driving pressure transmission units differently and forming the number of driving pressure units pressurizing each differently.

[0116] A driving pressure transmission unit may be formed on a membrane provided separately from the channel plate, and FIG. 13 is a drawing for explaining the joint structure of the channel plate and the membrane.

[0117] On the back surface of the channel plate (500), channels (C1 to C6) through which flow proceeds are formed, and connecting portions (H1 to H8) are formed in the form of holes.

[0118] When the above membrane (580) is attached to the back surface of the channel plate (500), specific channels (C1, C2) are connected to each other by the first driving pressure transmission unit (510a), thereby forming one channel composed of channel C1, the first driving pressure transmission unit (510a), and channel C2.

[0119] In particular, in the present invention, the function of the pump can be performed by arranging the driving pressure unit (335) so that the direction of movement and the longitudinal direction of the driving pressure transmission unit (510) are aligned with each other.

[0120] The driving pressure transmission unit (510) connects two channels among the plurality of channels (367) formed on the back surface of the channel plate (500). In addition, the driving pressure transmission unit (510) is provided at a position corresponding to the ring body (336) to perform channel and pumping functions. That is, according to the present invention, only a part of the originally single channel (367) is changed to a driving pressure transmission unit made of an elastomer material and a pumping operation is performed, so that a more accurate amount of culture medium can be supplied than when the entire channel is changed to a membrane.

[0121] The first culture medium of the first culture medium reservoir (541) is provided to the first type cells of the cell culture chip (100) through the first connection part (H1), the first channel (C1), the first driving pressure transmission part (510a), the second channel (C2), and the third connection part (H3).

[0122] And the first culture solution that has passed through the first type of cells of the cell culture chip (100) is recovered back into the first culture solution storage (541) through the fourth connecting portion (H4), the fifth channel (C5), and the second connecting portion (H2).

[0123] And the second culture medium of the second culture medium storage (542) is provided to the second type cells of the cell culture chip (100) through the fifth connection part (H5), the third channel (C3), the second driving pressure transmission part (510b), the fourth channel (C4), and the seventh connection part (H7).

[0124] And the second culture solution that has passed through the second type cells of the cell culture chip (100) is recovered back into the second culture solution storage (542) through the eighth connecting portion (H8), the sixth channel (C6), and the sixth connecting portion (H6).

[0125] Figures 14A to 14C are drawings for explaining the pumping operation for supplying and recovering culture medium.

[0126] As shown in FIGS. 14A to 14C, the driving pressure part (335) of the ring body (336) moves while pressurizing the driving pressure transmission part (510) formed in the channel plate (500) to change the shape of the driving pressure transmission part (510), thereby pumping the culture solution.

[0127] According to the present invention, a culture solution is supplied to a cell culture chip (100), and different culture solutions can be supplied simultaneously to two or more cells in the cell culture chip (100) by a drive transmission unit (510) that is pressurized by a drive pressure unit (335) of a cell solution supply device (300) and performs a pumping operation.

[0128] In addition, according to the present invention, in order to supply two or more culture media at different speeds within the cell culture chip (100), the internal cross-sectional areas of the two driving transmission units (510) are formed differently, thereby supplying the culture media to two or more cells within the cell culture chip (100) at different speeds.

[0129] In addition, according to the present invention, a cell culture chip (100), a chip holder (200), and a new cell culture device (300) are connected without requiring separate tubing work, thereby solving the problem of the supply amount of culture solution differing from the setting according to the tubing length, and has the advantage of simple initial setting.

[0130] Next, another embodiment of a culture solution supply device for a cell culture chip will be described with reference to FIGS. 15 and 16.

[0131] Referring to Fig. 15, the culture solution supply device for a cell culture chip according to the present embodiment has a structure in which a culture solution supply unit comprising a base (320) including a gear part (330) and a top cover (360) are stacked inside a case part (600). However, it is preferable that only one rotary drive part (312) be provided at the lowest part.

[0132] Specifically, the rotary shaft (319) connected to the rotary drive unit is provided to penetrate the base, gear unit, and top cover, and by driving multiple gear units at once with one rotary shaft, the structure is simplified and efficient operation is enabled.

[0133] The above-mentioned rotational axis can be coupled through a coupling device such as a bearing. In addition, it is preferable that the plurality of bases (320) are fixed to the case portion by a coupling mechanism such as a screw (610).

[0134] Referring to Fig. 16, a plurality of through holes (650) are formed in each layer of the case portion (600), and an integrated chip including a chip and a channel plate can be mounted through the through holes (650). The channel plate is of a type that accommodates a chip without a chip holder and a type in which a chip is accommodated in a chip holder and mounted on the channel plate.

[0135] Specifically, a plurality of integrated chips (800) including cell culture chips can be first mounted on the integrated chip mounting portion (700).

[0136] In the above-mentioned integrated chip mounting portion (700), a plurality of integrated chip receiving portions (710) capable of mounting integrated chips are formed with the same number of layers and height as the case portion (600), and the integrated chip is detachably coupled inside the integrated chip receiving portion.

[0137] Referring to Fig. 17A, the integrated chip receiving portion is provided with an integrated chip coupling portion (733) and an integrated chip injection portion (731).

[0138] The above-mentioned integrated chip joint portion has a hook-shaped end to engage with the protruding end of the integrated chip to prevent the detachment of the integrated chip. The above-mentioned integrated chip joint portion is provided so as to be rotatable by a rotational axis located at the center, and an elastic member (734) such as a spring or rubber for providing restoring force is connected to the upper end, and a release button joint portion (735) is provided at the lower end so as to be rotatable to a release button rod (752) connected to a release button (750).

[0139] A plurality of integrated chip coupling parts vertically stacked on one release button bar (752) connected to the above release button (750) are combined so that all of the stacked integrated chips can be released at once with one release button.

[0140] The above-mentioned integrated chip injection unit serves to push the integrated chip forward when disengaging, and the pushing power may be based on an elastic member such as a pneumatic cylinder, a hydraulic cylinder, or a spring.

[0141] Referring to Fig. 17B, when an integral chip is to be injected, pressing the release button (750) causes the integral chip coupling part to rotate by the rotation shaft, releasing the hook connection with the integral chip. At this time, the integral chip injection part pushes the integral chip forward, so that the integral chip is injected forward.

[0142] After inserting and mounting the integrated chip into each layer of the integrated chip mounting part (700), when the integrated chip mounting part is pressed against the case part (600) and the release button is pressed, the integrated chips of each layer move through the through hole at once and are installed in the culture solution supply device for the cell culture chip, so that the integrated chip can be installed very simply and efficiently.

[0143] The cell culture chips provided within the integrated chip installed on each layer of the culture medium supply device for the cell culture chip can be provided with different types of cells, such as liver and kidney. In addition, the rotation speed can be appropriately changed by varying the gear ratio of the gear part for each layer, and the shape of the driving pressure transmission part can be varied to provide a culture medium supply structure optimized for specific cells.

[0144] In addition, each layer of the culture medium supply device for the cell culture chip can supply a different type of culture medium appropriate for a specific cell, and the culture medium movement speed can be optimized by making the gear ratio or the shape of the driving pressure transmission unit different for each layer.

[0145] Meanwhile, referring to FIG. 18, the chip holder mounting portion (700) may be provided in a cylindrical shape with a hollow portion formed therein, and a case portion (600) may be inserted into the hollow portion.

[0146] According to this embodiment, since the chip holders provided in the chip holder mounting portion (700) can be moved and inserted simultaneously through the through holes of the case portion (600) facing each other, work efficiency is drastically improved.

[0147] Next, the process of mounting an integrated chip on a culture solution supply device for a cell culture chip according to the present embodiment will be described with reference to FIGS. 19A to 19F.

[0148] According to Fig. 19A, the culture solution supply device for the cell culture chip according to the present embodiment is composed of multiple layers and is installed rotatably with respect to the base cover.

[0149] In addition, a hole is formed on the side into which a channel plate can be inserted, and the channel plate is inserted and mounted from the side as shown in FIGS. 18B to 18D. At this time, it is also possible to mount multiple channel plates at once using the chip holder mounting portion.

[0150] After all the channel plates are mounted on one side of the culture medium supply device for the cell culture chip, the upper part can be rotated with respect to the base cover, and then the channel plates can be mounted on the adjacent side again.

[0151] After all the channel plates are installed, the internal gears can be rotated to supply the culture medium to each layer at different speeds and amounts.

[0152]

[0153] FIG. 20 is a diagram illustrating a configuration of a cell culture system, an analysis device, and a user terminal according to one embodiment. Referring to FIG. 20, an automated cell culture process according to one embodiment can be implemented through a cell culture system (1000), an analysis device (2000), and a user terminal (3000).

[0154] The cell culture system (1000) is a device comprising various modules for controlling cell seeding, culture, and the biological microenvironment, and can provide an automated cell culture process. The cell culture system (1000) can accurately seed and culture cells through a seeding kit and platform, and can include a biological microenvironment control module for this purpose.

[0155] The analysis device (2000) may be an external device capable of performing various analyses on cells cultured in the cell culture system (1000). It may monitor cells cultured in the cell culture system (1000) in real time, collect various biological data such as cell status, response, and growth patterns, and provide information necessary for research. The analysis device (2000) may be closely linked with the cell culture system (1000) to provide an automated analysis process.

[0156] The user terminal (3000) can control the operation of the cell culture system (1000) and the analysis device (2000), and can provide an interface for monitoring data. The researcher can issue commands to the cell culture system (1000) through the user terminal (3000) and check data generated through the analysis device (2000) in real time. In addition, the user terminal (3000) can store and analyze data generated during the cell culture and analysis process, and perform functions such as visually representing the results or generating reports.

[0157]

[0158] Figures 21 to 24 are drawings for explaining the configuration of a cell culture system according to one embodiment. Referring to Figure 21, the cell culture system (1000) may include a seeding kit (1100), a platform (1200), a cartridge (1300), and a biomicroenvironment control module (1400).

[0159] A seeding kit (1100) is a device for seeding cells in the initial stage of a cell culture system, and can perform an operation to accurately and uniformly distribute cells onto specific layers of a cartridge in which cells will be cultured. Referring to (a) of Fig. 23, the seeding kit (1100) can perform a seeding operation with layers mounted on the main body.

[0160] The platform (1200) may include a space capable of combining cartridges (1300) and may perform an operation of circulating a culture medium through the cartridges (1300). The platform (1200) may securely secure the cell-seeded cartridges (1300) and constantly circulate necessary nutrients to support cell growth in an optimal environment. Referring to (b) of FIG. 23, the platform (1200) may have an internal space capable of accommodating multiple cartridges and may include an interface for controlling the device.

[0161] Referring to FIG. 22, the platform (1200) may include at least one processor (1210), a communication module (1220) communicatively connected to an external device, a cartridge mounting module (1230), a driving module (1240), a sensor module (1250), and an internal environment control module (1260). Meanwhile, the platform (1200) described in the present specification may have a configuration corresponding to the cell culture device (10) described above with reference to FIGS. 1 to 9. That is, the platform (1200) may include a configuration such as a cell culture chip (100), a chip holder (200), a culture solution supply device (300), and a channel plate (500), and detailed descriptions thereof have been described above, so redundant descriptions thereof will be omitted.

[0162] The cartridge (1300) is a device in which cells are seeded and cultured, and is characterized by including multiple layers (a first layer, a second layer, and a third layer). Each layer of the cartridge (1300) provides specific environmental conditions and can create a physical environment necessary for cell growth and development. Referring to FIG. 24, the cartridge (1300) may include the drive transmission unit (510) described with reference to FIGS. 11 to 14, and may include a reservoir (540) for storing a culture medium.

[0163] The bio-microenvironment control module (1400) is a device that controls the environment inside the platform (1200) where cell culture is performed, and can perform an operation to maintain bio-environment variables such as temperature, humidity, and carbon dioxide concentration in an optimized state.

[0164]

[0165] FIG. 25 is a diagram illustrating a method for culturing cells using a cell culture supply device according to one embodiment. Referring to FIG. 25, a cell automation process according to one embodiment may include a step of seeding and stabilizing cells in a cartridge (S110), a step of coupling the cartridge to a platform (S120), a step of circulating a culture medium in a cartridge coupled to the platform (S130), a step of replacing and sampling the culture medium (S140), a step of detaching the cartridge from the platform (S150), and a step of performing an analysis on the cells (S160).

[0166]

[0167] FIG. 26 is a drawing for detailing the cell seeding and stabilization steps according to one embodiment.

[0168] Referring to FIG. 26, the cell seeding and stabilization step according to one embodiment may include a step of combining a middle layer to a seeding kit (S210), a step of injecting a cell suspension, ECM, and culture medium into a first seeding area of ​​the middle layer (S220), a step of creating a cell culture environment and stabilizing cells for a predetermined time (S230), a step of flipping the middle layer (S240), a step of injecting a cell suspension, ECM, and culture medium into a second seeding area of ​​the middle layer (S250), and a step of creating a cell culture environment and stabilizing cells for a predetermined time (S260).

[0169] The cartridge (1300) may include a plurality of layers. The plurality of layers may be divided into a first layer, a second layer, and a third layer, wherein the first layer may be divided into an upper layer, the second layer may be divided into a middle layer, and the third layer may be divided into a lower layer. In addition, the first layer may be divided into a top layer, the second layer may be divided into a middle layer, and the third layer may be divided into a bottom layer. The second layer may be positioned between the first layer and the third layer.

[0170] The seeding kit (1100) may include a module capable of accommodating the second layer (hereinafter, referred to as a middle layer), and a seeding operation may be performed on the middle layer disposed in the module.

[0171] The middle layer may include a plurality of seeding regions. The middle layer may include a first seeding region and a second seeding region, wherein the first seeding region may be a region formed on a first surface of the middle layer, and the second seeding region may be a region formed on a second surface of the middle layer. Here, the second surface may be a surface facing opposite to the first surface.

[0172] The step of combining the middle layer with the seeding kit (S210) may include a step of combining the middle layer so that the first seeding area faces in the opposite direction to the ground. The step of flipping the middle layer (S240) may include a step of flipping the middle layer so that the second seeding area faces in the opposite direction to the ground. In this way, by injecting the cell suspension, ECM, and culture medium while the seeding area of ​​the middle layer faces in the opposite direction to the ground, cell seeding can be stably performed.

[0173] More specifically, in the past, there was a problem that cell suspensions, etc. were unevenly distributed during the cell seeding process or seeding was only performed on a specific surface, resulting in inconsistency in the culturing process. Therefore, in the cell automation process according to the present invention, cell suspensions, etc. are injected into the first seeding area and the second seeding area (i.e., both areas of the middle layer) through an operation of flipping the middle layer, thereby enabling cells to be uniformly distributed on both sides of the middle layer.

[0174] Although not shown in the drawing, the steps of stabilizing the cells (S230, S260) may each include a step of mounting a cover in a space accommodating the middle layer to block the middle layer from the external environment.

[0175]

[0176] Figure 27 is a drawing for explaining a loading step according to one embodiment.

[0177] Referring to FIG. 27, another loading step in one embodiment may include a step of removing the culture medium after the stabilization operation is completed (S310), a step of dismantling the middle layer from the seeding kit (S320), a step of mounting the middle layer on a cartridge (S330), and a step of dispensing the culture medium into a reservoir of the cartridge (S340).

[0178] The step (S310) of removing the culture medium may be performed after the stabilization operation is completed and cell stabilization is achieved, thereby removing any remaining culture medium within the seeding kit. This step may be performed to prevent the culture medium from leaking out when the middle layer is mounted on the cartridge in a subsequent step. By preventing the culture medium from leaking out, the experimental environment can be maintained cleanly.

[0179] The step (S320) of disassembling the middle layer from the seeding kit is an operation of disassembling the middle layer coupled to the seeding kit, and may be an operation of opening the cover of the seeding kit and taking out the middle layer placed inside.

[0180] The step of mounting the middle layer on the cartridge (S330) may include an operation of placing a third layer (bottom layer) in the internal space of the cartridge, an operation of placing the middle layer on the third layer, and an operation of placing the first layer (top layer) on the middle layer.

[0181] The step of dispensing the culture solution (S340) may be an operation of dispensing an appropriate amount of culture solution required for cell culture into a reservoir within the cartridge, thereby providing nutrients necessary for cell growth and survival. By dispensing a predetermined amount of culture solution into the reservoir's receiving space, appropriate nutrients can be supplied to cells seeded in the middle layer.

[0182]

[0183] FIG. 28 is a drawing illustrating a step of circulating a culture medium in a cartridge within a platform according to one embodiment.

[0184] Referring to FIG. 28, the culture medium circulation step according to one embodiment may include a step of controlling a driving module based on a setting value determined in a predetermined manner (S410), a step of creating a biological microenvironment (S420), a step of monitoring and controlling an internal environment (S430), and a step of monitoring the state of cells within a cartridge (S430).

[0185] The drive module according to one embodiment may correspond to the gear unit (330) described above with reference to FIGS. 6 to 8, and the step (S410) of controlling the drive module may be performed based on the operation of the gear unit (330).

[0186] The step of creating a biological microenvironment (S420) and the step of monitoring and controlling the internal environment (S430) may be an operation of controlling the environment within the platform (1200) so that cell culture can proceed smoothly. More specifically, it may mean an operation of maintaining biological environmental variables such as temperature, humidity, and carbon dioxide within the platform (1200) in an optimized state. The above-described operation may be performed by the biological microenvironment control module (1400).

[0187] The platform (1200) may include at least one sensor. At least one processor (1210) of the platform (1200) may monitor the state of cells within the cartridge based on the at least one sensor. A plurality of cartridges may be arranged within the platform (1200), and at least one processor (1210) may perform a monitoring operation for each of the plurality of cartridges based on at least one sensor each arranged at a location corresponding to the plurality of cartridges.

[0188] More specifically, the plurality of cartridges may include a first cartridge, a second cartridge, and a third cartridge, and at least one sensor may include a first sensor, a second sensor, and a third sensor. Here, at least one processor (1210) may monitor the status of the first cartridge based on the first sensor, the status of the second cartridge based on the second sensor, and the status of the third cartridge based on the third sensor. In this way, by performing individual monitoring on a cartridge-by-cartridge basis, problems occurring in a specific cartridge can be detected and responded to in real time.

[0189]

[0190] FIG. 29 is a drawing for explaining a step of replacing and sampling a culture medium according to one embodiment.

[0191] Referring to FIG. 29, the culture medium replacement and sampling step according to one embodiment may include a step of opening a reservoir cover of a cartridge (S510), a step of sampling a culture medium in a reservoir at a predetermined rate (S520), a step of injecting a new culture medium into the reservoir at the predetermined rate (S530), and a step of analyzing the sampled culture medium (S540).

[0192] The step of opening the reservoir cover (S510) is an operation for sampling and replacing the culture solution contained in the reservoir, and the step of sampling the culture solution in the reservoir (S520) may be an operation for replacing a certain portion of the culture solution in consideration of the possibility of contamination or depletion of nutrients.

[0193] The step (S530) of injecting new culture medium into the reservoir may be an operation to replace the sampled culture medium. This operation ensures that cells can continue to grow in an optimal culture environment. By replacing only a certain percentage of the culture medium, rather than the entire volume, a stable environment required for the cells can be maintained. Here, the predetermined ratio may be set differently depending on the type and conditions of the experiment.

[0194] More specifically, since the method of replacing the entire culture medium has the problem of causing stress to the cells and lowering the cell survival rate, the present invention can prevent a rapid change in the cell environment and maintain a stable culture environment by replacing only a certain percentage of the culture medium in the reservoir.

[0195] The sampled culture fluid analysis step (S540) can perform an electrochemical analysis of the culture fluid based on a sensor. This analysis can monitor the condition of the culture fluid, whether it is contaminated, and the concentration of nutrients, and can also determine the progress of the cell culture.

[0196]

[0197] FIG. 30 is a diagram illustrating steps for performing analysis on cells according to one embodiment.

[0198] Referring to FIG. 30, the cell analysis step according to one embodiment may include a step of removing a culture medium in a reservoir (S610), a step of separating a middle layer from a cartridge (S620), a step of combining the middle layer with a seeding kit (S630), a step of performing cell staining in the seeding kit according to a protocol (S640), and a step of separating the middle layer from the seeding kit after staining (S650).

[0199] The step of removing the culture medium within the reservoir (S610) may be an operation to remove the remaining culture medium within the reservoir, thereby preparing the middle layer for safe separation from the cartridge. By removing the culture medium in advance, the culture medium can be prevented from leaking out during the middle layer separation process, and the working environment can be maintained cleanly.

[0200] The step (S620) of separating the middle layer from the cartridge may be an operation of removing the plate of the cartridge and removing the middle layer. This operation safely separates the middle layer for analysis of cells cultured within the cartridge, and through this operation, the middle layer can be separated to perform staining and analysis operations.

[0201] The step of combining the middle layer with the seeding kit (S630) may be an operation of placing the middle layer in the internal space of the seeding kit similar to the seeding step, and may be an operation of stably combining and placing the middle layer within the seeding kit for cell staining.

[0202] The step (S640) of performing cell staining within the seeding kit according to the protocol may be an operation of performing cell staining on the first seeding area and the second seeding area of ​​the middle layer, respectively, and through such staining operation, cell structures or specific intracellular components can be visually distinguished, thereby assisting in performing the subsequent analysis process more accurately. Meanwhile, according to an embodiment, the step (S640) of performing cell staining may include an operation of performing staining on only one of the first seeding area and the second seeding area of ​​the middle layer, or may include an operation of performing staining on both the first seeding area and the second seeding area.

[0203] The step (S650) of separating the middle layer from the seeding kit after dyeing may be an operation of opening the cover of the seeding kit and separating the middle layer after dyeing is completed. Through this operation, the middle layer can be separated and moved to an analysis device (2000), whereby the condition or response of the cells can be precisely analyzed.

[0204]

[0205] The features, structures, effects, etc. described in the embodiments above are included in at least one embodiment of the present invention, and are not necessarily limited to just one embodiment. Furthermore, the features, structures, effects, etc. exemplified in each embodiment can be combined or modified in other embodiments by those skilled in the art to which the embodiments pertain. Therefore, the contents related to such combinations and modifications should be construed as falling within the scope of the present invention.

[0206] In addition, although the above description focuses on the embodiments, these are merely examples and do not limit the present invention. Those skilled in the art to which the present invention pertains will appreciate that various modifications and applications not exemplified above are possible without departing from the essential characteristics of the present embodiment. In other words, each component specifically shown in the embodiments can be modified and implemented. In addition, differences related to such modifications and applications should be interpreted as being included within the scope of the present invention defined in the appended claims.

Claims

1. A method for culturing cells using a cell culture supply device, Step of seeding cells into cartridges; A step of combining the above cartridge to the platform; A step of injecting a culture medium into the cartridge coupled to the platform; and A step of circulating the culture medium within the platform; including: The step of seeding cells into the above cartridge is: A step of combining the middle layer included in the above cartridge into a seeding kit; a step of seeding cells in different areas of the middle layer; A method for culturing cells using a cell culture supply device.

2. In paragraph 1, The above different regions include a first seeding region and a second seeding region, The step of seeding the above cells is: a step of seeding cells in the first seeding area of ​​the middle layer; and comprising a step of seeding cells in the second seeding area of ​​the middle layer; A method for culturing cells using a cell culture supply device.

3. In paragraph 2, The step of seeding cells into the above cartridge is: Further comprising the step of flipping the middle layer, A method for culturing cells using a cell culture supply device.

4. In paragraph 3, After the step of seeding cells in the first seeding area is performed, the step of flipping the middle layer is performed, After the step of flipping the middle layer is performed, the step of seeding cells in the second seeding area is performed. A method for culturing cells using a cell culture supply device.

5. In paragraph 2, The step of seeding cells in the first seeding area is performed in a state where the first seeding area faces in the opposite direction to the ground, The step of seeding cells in the second seeding area is performed in a state where the second seeding area faces in the opposite direction to the ground. A method for culturing cells using a cell culture supply device.

6. In paragraph 2, The step of seeding cells in the first seeding area comprises the step of injecting at least one of a cell suspension, ECM, and culture medium into the first seeding area, The step of seeding cells in the second seeding area comprises the step of injecting at least one of a cell suspension, ECM, and culture medium into the second seeding area. A method for culturing cells using a cell culture supply device.

7. In paragraph 2, The step of combining the above cartridge to the platform is: A step of removing the culture medium from the seeding kit after the cell stabilization operation is performed; A step of separating the middle layer from the seeding kit; a step of combining the middle layer to the cartridge; and A step of dispensing a culture medium into the reservoir of the cartridge; comprising; A method for culturing cells using a cell culture supply device.

8. In paragraph 2, A method for culturing cells using a cell culture supply device, Further comprising a step of replacing the culture medium in the reservoir, The step of replacing the culture medium in the reservoir is as follows: A step of sampling the culture medium in the reservoir at a predetermined ratio; and A step of injecting a new culture medium into the reservoir in the predetermined ratio; A method for culturing cells using a cell culture supply device.

9. In paragraph 2, A method for culturing cells using a cell culture supply device, Including additional cell staining steps, The above cell staining step is, A step of separating the middle layer from the cartridge; a step of combining the above middle layer with the above seeding kit; and A step of performing cell staining in the seeding kit according to the protocol; including; A method for culturing cells using a cell culture supply device.

10. In paragraph 2, A method for culturing cells using a cell culture supply device, Further comprising a step of monitoring the state of cells within the cartridge, wherein the cartridge comprises a first cartridge, a second cartridge and a third cartridge, The cell status monitoring step within the above cartridge is: A step of monitoring the status of the first cartridge based on the first sensor; A step of monitoring the status of the second cartridge based on a second sensor; and A step of monitoring the status of the third cartridge based on a third sensor; A method for culturing cells using a cell culture supply device.

11. In paragraph 10, The first sensor is positioned at a position corresponding to the first cartridge, the second sensor is positioned at a position corresponding to the second cartridge, and the third sensor is positioned at a position corresponding to the third cartridge. A method for culturing cells using a cell culture supply device.

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