Cell culture chamber, culture device and culture process

Abstract

This application relates to the field of cell culture technology, specifically to a cell culture chamber, culture device, and culture process. The cell culture chamber includes a chamber body, a cover, and a carrier. The chamber body has a culture cavity, and the cover is detachably connected to the chamber body. The carrier extends along a first direction and is connected to the cover. The carrier is located within the culture cavity and has an inner cavity and multiple through holes. The inner cavity communicates with the culture cavity, and each through hole penetrates the cavity wall and communicates with the inner cavity. The culture device includes a frame, a control unit, a drive unit, and the aforementioned cell culture chamber. The drive unit is connected to the frame and electrically connected to the control unit. The chamber body is drive-connected to the drive unit, and the chamber body is configured to rotate around the first direction under the drive unit's influence. The cell culture process utilizes the aforementioned cell culture device to culture cells. The cell culture chamber, culture device, and culture process of this application can improve the uniformity of cell spheroid culture size.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell culture, in particular to a cell culture cabin, a culture device and a culture process. BACKGROUND

[0002] In the field of cell culture technology, in order to simulate the three-dimensional microenvironment in vivo, various three-dimensional shaping methods are usually used to culture cell spheres. However, the cell spheres formed by the existing methods are solid spheres, and after the diameter of the sphere increases with the culture process, it is difficult for nutrients and oxygen to penetrate into the interior of the cell sphere, and it is also difficult for cell secretion products to penetrate out, resulting in insufficient uniformity of the formed size of the cell sphere. SUMMARY

[0003] The present application aims to at least solve one of the technical problems existing in the prior art. To this end, the present application provides a cell culture cabin capable of improving the uniformity of the size of the cell sphere culture.

[0004] The present application also provides a culture device having the above-mentioned cell culture cabin.

[0005] The present application also provides a culture process for culturing cells using the above-mentioned cell culture device.

[0006] The cell culture cabin according to the embodiments of the present application comprises a cabin body, a cover body and a carrier;

[0007] The cabin body is provided with a culture cavity;

[0008] The cover body is detachably connected to the cabin body;

[0009] The carrier is arranged along a first direction, the carrier is connected to the cover body, the carrier is located in the culture cavity, the carrier is provided with an inner cavity and a plurality of through holes, the inner cavity is communicated with the culture cavity, each through hole penetrates the cavity wall of the inner cavity, and each through hole is communicated with the inner cavity.

[0010] The cell culture cabin according to the embodiments of the present application has at least the following beneficial effects: the culture cavity is used to accommodate cell suspension, during cell culture, the carrier is driven to rotate around the first direction, the cells in the cell suspension are aggregated on the outer peripheral wall of the carrier, and then grow to form cell spheres, during the cell culture process, nutrients and oxygen can be transported to the central region of the cell sphere through the inner cavity and the through holes, so as to ensure the growth of the cell sphere, and the secretion products of the cell sphere can also be transported into the culture cavity through the through holes and the inner cavity, thereby ensuring the healthy growth and development of the cell sphere, which is beneficial to ensure the uniformity of the formed size of the cell sphere and can prolong the cultivation time of the cell sphere.

[0011] According to some embodiments of the present application, the carrier comprises a hydrophilic culture section and a hydrophobic isolation section connected in sequence along the first direction, and the hydrophilic culture section is provided with an inner cavity and a through hole.

[0012] According to some embodiments of the present application, the hydrophobic isolation section is provided with an inner cavity and a through hole.

[0013] According to some embodiments of the present application, the length of the hydrophilic culture section along the first direction is 0.1mm to 1mm.

[0014] And / or, the length of the hydrophobic isolation section along the first direction is 1mm to 5mm.

[0015] According to some embodiments of the present application, the cell culture cabin includes a plurality of carriers, each carrier being connected with the cover, and the carriers being arranged at intervals.

[0016] According to the cell culture device of the embodiments of the present application, the cell culture device comprises a frame, a control unit, a driving unit and the cell culture cabin of any of the above embodiments.

[0017] The driving unit is connected with the frame, and the driving unit is electrically connected with the control unit.

[0018] The cabin of the cell culture cabin is drivingly connected with the driving unit, and the cabin is configured to be driven by the driving unit to rotate around the first direction.

[0019] According to the culture device of the embodiments of the present application, at least the following beneficial effects are achieved: the driving unit can drive the cell culture cabin to rotate, the control unit is used to control the start, stop, rotation speed and other operations of the driving unit, the controllable cell culture operation is realized, and the cell culture efficiency is improved.

[0020] According to some embodiments of the present application, the driving unit comprises a driving member and at least two rotating members, each rotating member being arranged in sequence along a second direction, the driving member being connected with the frame and being drivingly connected with each rotating member for driving each rotating member to rotate around its own axis, the axis of each rotating member being parallel to the first direction, the cabin being stacked with adjacent two rotating members along a third direction, and the first direction, the second direction and the third direction being perpendicular to each other in pairs.

[0021] According to the culture process of the embodiments of the present application, the culture process comprises two steps of culture preparation and agglomerated growth.

[0022] In the culture preparation, cell suspension is added into the culture cavity until the cell suspension immerses the carriers.

[0023] In the agglomerated growth, the power output of the driving unit is controlled by the control unit, so that the driving unit drives the cell culture cabin to rotate around the first direction, the cells in the cell suspension are agglomerated on the outer peripheral wall of the carrier, and the cells grow to form cell balls.

[0024] According to the culture process of the embodiment of the present application, the carrier has an inner cavity and a through hole, the cells are aggregated on the outer peripheral wall of the carrier, the nutrients and oxygen can be transported to the central region of the cell ball through the inner cavity and the through hole, and the products of the cell ball can also be transported into the culture cavity through the through hole and the inner cavity, which ensures the stability and uniformity of the microenvironment of the cell ball everywhere, and is beneficial to improve the uniformity of the culture size of the cell ball.

[0025] According to some embodiments of the present application, in the culture preparation step, the hydrophobic material is coated on the outer peripheral wall of the carrier at intervals, so that the hydrophobic isolation section and the hydrophilic culture section are formed on the carrier in sequence along the first direction.

[0026] In the aggregation growth step, the cells are aggregated on the outer peripheral wall of the hydrophilic culture section.

[0027] According to some embodiments of the present application, the rotation speed of the carrier ranges from 10 r / min to 200 r / min.

[0028] Additional aspects and advantages of the present application will be given in part in the following description, will become apparent from the following description, or will be learned by practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0029] The present application will be further described below in combination with the drawings and embodiments, in which:

[0030] Figure 1 It is an exploded view of the cell culture cabin of the embodiment of the present application;

[0031] Figure 2 It is a side view of the cell culture cabin of the embodiment of the present application;

[0032] Figure 3 It is Figure 2 the cross-sectional view at A-A;

[0033] Figure 4 It is a schematic view of the carrier for culturing the cell ball of the embodiment of the present application;

[0034] Figure 5 It is a schematic view of the culture device of the embodiment of the present application;

[0035] Figure 6 It is a side view of the culture device of the embodiment of the present application.

[0036] Reference signs: cabin body 110, culture cavity 111, cover body 120, carrier 130, inner cavity 131, through hole 132, hydrophilic culture section 133, hydrophobic isolation section 134;

[0037] frame body 210, control unit 220, driving unit 230, driving member 231, rotating member 232;

[0038] Cell sphere 310. Detailed Implementation

[0039] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0040] In the description of this application, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0041] In the description of this application, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0042] In the description of this application, unless otherwise expressly defined, terms such as "setup," "installation," and "connection" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this application in conjunction with the specific content of the technical solution.

[0043] In the description of this application, the terms "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0044] The embodiments of this application are described below with reference to the accompanying drawings:

[0045] refer to Figures 1 to 4According to the cell culture cabin provided by the embodiment of the present application, the cell culture cabin comprises a cabin body 110, a cover body 120 and a carrier 130. The cabin body 110 is provided with a culture cavity 111. The cover body 120 is detachably connected with the cabin body 110. The carrier 130 is arranged in extension along a first direction. The carrier 130 is connected with the cover body 120. The carrier 130 is located in the culture cavity 111. The carrier 130 is provided with an inner cavity 131 and a plurality of through holes 132. The inner cavity 131 is communicated with the culture cavity 111. Each through hole 132 penetrates the cavity wall of the inner cavity 131. Each through hole 132 is communicated with the inner cavity 131. In this way, the outer peripheral wall of the carrier 130 is used for carrying the cell aggregation and shaping. The carrier 130 can transport nutrients and oxygen to the center of the cell sphere 310 through the inner cavity 131 and the through holes 132. In addition, the inner cavity 131 and the through holes 132 can also ensure the exudation of cell secretion products, which helps to prolong the culture and test time of the cell sphere 310, thereby improving the uniformity of the shaped size of the cell sphere 310.

[0046] Specifically, the cabin body 110 is a cavity structure with an open top. The cabin body 110 encloses to form the culture cavity 111. When the cells are cultured, the culture cavity 111 can accommodate the cell suspension. The cover body 120 and the cabin body 110 can be detachably matched by using threaded connection, buckle connection or the like. The carrier 130 is arranged in extension along the first direction. For example, the carrier 130 can be a cylindrical structure with an axis extending along the first direction. The diameter of the carrier 130 can range from 100 µm to 600 µm. The side of the cover body 120 facing the cabin body 110 can be circular, square or the like. Taking the circular shape as an example, the carrier 130 can be connected to the center of the cover body 120. The carrier 130 is located in the culture cavity 111. When the cover body 120 is connected with the cabin body 110 to close the culture cavity 111, the outer peripheral wall of the carrier 130 has a spacing with the inner cavity 131 wall of the culture cavity 111. In addition, the carrier 130 can be immersed in the cell suspension accommodated in the culture cavity 111.

[0047] The carrier 130 can be made of hydrophilic materials, such as hydrophilic polymers like polysulfone and polyvinyl chloride. During cell culture, the cell culture chamber is driven to rotate around a first direction, causing the cells in the cell suspension to remain suspended. The suspended cells contact the carrier 130 and aggregate on the outer wall of the carrier 130. During the continuous rotation of the culture chamber, the aggregated cells gradually proliferate and differentiate to form cell spheres 310. Because the carrier 130 has an inner cavity 131 and through holes 132, it has a hollow porous structure. When the carrier 130 is immersed in the cell suspension and rotates, nutrients and oxygen can enter the interior of the carrier 130 through the inner cavity 131 and permeate to the center of the cell spheres 310 through the multiple through holes 132, ensuring that all parts of the cell spheres 310 receive sufficient nutrients and oxygen. In addition, metabolic products such as exosomes, proteins, and microvesicles produced by cell secretion can also be quickly discharged into the culture chamber 111 through the through-hole 132 and the inner cavity 131, so that the culture conditions inside the cell ball 310 remain stable and uniform, which is beneficial to improving the uniformity of the cell ball 310's forming size.

[0048] It should also be noted that a stable culture microenvironment can reduce cell apoptosis caused by nutrient deficiency, insufficient oxygen, or accumulation of metabolic products, which helps to prolong the culture time of cell spheres 310, enabling them to maintain a healthy physiological state over a longer period. This provides a reliable cell model and a longer observation window for drug screening and developmental biology research.

[0049] refer to Figures 1 to 4 In some embodiments, the carrier 130 includes a hydrophilic culture section 133 and a hydrophobic isolation section 134 connected together. The hydrophobic isolation section 134 and the hydrophilic culture section 133 are arranged sequentially along a first direction. The hydrophilic culture section 133 is provided with an inner cavity 131 and a through hole 132. Through the barrier of the hydrophobic isolation section 134, multiple cell spheres 310 can be cultured simultaneously and independently, which helps to further improve the uniformity of the cultured cell spheres 310 size.

[0050] Specifically, the hydrophobic isolation segment 134 can be made of a hydrophobic material, or the carrier 130 can be made of a hydrophilic material. The hydrophobic isolation segment 134 is formed by coating the carrier 130 with hydrophobic material at intervals along the first direction. Both the hydrophilic culture segment 133 and the hydrophobic isolation segment 134 can be set as multiple segments, and the hydrophilic culture segment 133 and the hydrophobic isolation segment 134 are arranged alternately along the first direction. The cover 120 is connected to the hydrophobic isolation segment 134 at the end of the carrier 130 to provide growth space for the cell sphere 310 near the cover 120. During cell culture, the cell suspension comes into contact with the hydrophilic culture section 133. Cells in the suspension aggregate on the outer wall of the hydrophilic culture section 133 and can transport nutrients and oxygen, as well as cell products, through the lumen 131 and through-holes 132 on the hydrophilic culture section 133. The hydrophobic isolation section 134, with its hydrophobic properties, is used to block different hydrophilic culture sections 133, ensuring that multiple cell spheres 310 can be cultured simultaneously on the carrier 130, while avoiding mutual interference between the cell spheres 310. Because the cell spheres 310 are cultured and grown on the isolated hydrophilic culture sections 133, their growth state is more uniform, which helps to further improve the uniformity of the cell sphere size and improves the efficiency of cell culture.

[0051] refer to Figures 1 to 4 In some embodiments, the hydrophobic isolation section 134 is provided with an inner cavity 131 and a through hole 132. That is, along the first direction, the inner cavity 131 connects the hydrophilic culture section 133 and the hydrophobic isolation section 134. Through the through hole 132 of the hydrophobic isolation section 134, it is beneficial to improve the flow efficiency of nutrients, oxygen and cell products, thereby further ensuring the uniformity of cell sphere 310 culture.

[0052] Specifically, the carrier 130 extends along a first direction, and a cylindrical inner cavity 131 extending through the carrier 130 along the first direction is formed inside the carrier 130. The inner cavity 131 passes through both the hydrophilic culture section 133 and the hydrophobic isolation section 134, so that the internal space of the hydrophilic culture section 133 and the internal space of the hydrophobic isolation section 134 are connected. A plurality of through holes 132 are uniformly formed radially on the side wall of the hydrophilic culture section 133. The plurality of through holes 132 are evenly distributed around the axial direction (i.e., the first direction) of the hydrophilic culture section 133. For example, at least three through holes 132 are uniformly distributed circumferentially at the same axial height. A plurality of through holes 132 are also uniformly formed radially on the side wall of the hydrophobic isolation section 134, and the axial and circumferential distribution density can be consistent with that of the through holes 132 of the hydrophilic culture section 133. One end of each through hole 132 is connected to the through inner cavity 131, and the other end is connected to the culture chamber 111.

[0053] During cell culture, the culture chamber 111 contains a cell suspension. The cell culture chamber is driven to rotate around the first direction. During the rotation, the cell suspension in the culture chamber 111 flows with the rotation of the chamber 110, forming a stable rotating flow field. Due to the hydrophilic properties of the hydrophilic culture section 133, the cells in the cell suspension gradually gather and aggregate on the surface of the hydrophilic culture section 133. As the rotation culture continues, the aggregated cells gradually develop into cell spheres 310, which cover the outer wall of the hydrophilic culture section 133.

[0054] The transport pathway for nutrients and oxygen is as follows: A portion of the nutrients and oxygen in culture chamber 111 directly enters inner cavity 131, while the other portion enters inner cavity 131 through the through-hole 132 of hydrophobic isolation section 134. After entering inner cavity 131, they are transported along inner cavity 131 to the corresponding area of ​​hydrophilic culture section 133, and then through the through-hole 132 of hydrophilic culture section 133 to the portion of cell sphere 310 covering hydrophilic culture section 133, thus providing cell nutrient delivery to the side of cell sphere 310 covering hydrophilic culture section 133. Simultaneously, the cell spheres 310, encapsulating exosomes, proteins, microvesicles, and other metabolic products generated on one side of the hydrophilic culture section 133, flow into the inner cavity 131 through the through-holes 132 of the hydrophilic culture section 133. They then flow along the inner cavity 131 to the region corresponding to the hydrophobic isolation section 134, and are subsequently transported to the culture chamber 111 through the through-holes 132 of the hydrophobic isolation section 134. Alternatively, they can be directly transported to the culture chamber 111 through the portion connecting the inner cavity 131 and the culture chamber 111, thus achieving the transfer of metabolic products. This facilitates more efficient and complete penetration of nutrients and oxygen into the cell spheres 310, ensures the removal of cell analysis products, and extends the overall culture time of the cell spheres 310, allowing them to maintain a stable and healthy state over a longer period. This provides a more reliable and realistic cell model and a longer observation window.

[0055] refer to Figures 1 to 4 In some embodiments, the length of the hydrophilic culture segment 133 along the first direction is 0.1 mm to 1 mm, and / or the length of the hydrophobic isolation segment 134 along the first direction is 1 mm to 5 mm. By limiting the length range of the hydrophilic culture segment 133, the growth space of the cell spheres 310 can be restricted, which is beneficial to make the cell spheres 310 grow more uniformly in the culture chamber 111. By limiting the length of the hydrophobic isolation segment 134, the isolation of the growth of two adjacent cell spheres 310 can be taken into account, as well as ensuring the efficient flow of nutrients, oxygen and cell products.

[0056] For example, the length of the hydrophilic culture segment 133 along the first direction can be 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm or 1.0 mm, or fall within the range of any two of the above values.

[0057] The length of the hydrophobic isolation section 134 along the first direction can be 1 mm, 2 mm, 3 mm, 4 mm or 5 mm, or within the range of any two of the above values.

[0058] refer to Figures 1 to 4 In some embodiments, the cell culture chamber includes multiple carriers 130, each carrier 130 being connected to the cover 120, and the carriers 130 being arranged at intervals.

[0059] Specifically, multiple carriers 130 are disposed inside the culture chamber 111. Along the first direction, the multiple carriers 130 are connected to the side of the cover 120 facing the chamber 110, and the connection positions of the multiple carriers 130 can be arranged at intervals around the first direction. The spacing between adjacent carriers 130 is the same to ensure that the fluid environment of each carrier 130 within the culture chamber 111 is consistent. Each carrier 130 may have a hydrophilic culture section 133 and a hydrophobic isolation section 134 arranged sequentially. Both the hydrophilic culture section 133 and the hydrophobic isolation section 134 are provided with through holes 132. The hydrophobic isolation section 134 is connected to the cover 120. Each carrier 130 has an inner cavity 131 extending along the first direction, penetrating the hydrophilic culture section 133 and the hydrophobic isolation section 134. The through holes 132 communicate with the inner cavity 131. Simultaneous culture of cell spheres 310 using multiple carriers 130 can improve culture efficiency.

[0060] refer to Figures 1 to 6 According to the embodiments of this application, a cell culture apparatus is used to culture cells to form cell spheres 310. The cell culture apparatus includes a frame 210, a control unit 220, a drive unit 230, and a cell culture chamber as described in any of the above embodiments. The drive unit 230 is connected to the frame 210 and electrically connected to the control unit 220. The chamber 110 is rotatably connected to the drive unit 230 and is also driveably connected to the drive unit 230. The chamber 110 is configured to rotate around a first direction driven by the drive unit 230, which helps to ensure the stability of the rotation of the cell culture chamber and the controllability of the rotation speed, thereby helping to improve the uniformity of the cell sphere 310 culture. In addition, the drive unit 230 can drive multiple cell culture chambers to rotate simultaneously, thereby improving the cell culture efficiency.

[0061] Specifically, the drive unit 230 can drive the cell culture chamber to rotate around the first direction using methods such as roller drive, gear drive, and transmission belt drive. For example, the drive unit 230 includes a drive motor and two parallel rollers. The two ends of the rollers are connected to the frame 210 through bearings. The drive motor drives the rollers to rotate through a transmission belt or transmission chain. The chamber 110 is in contact with the outer surfaces of the two rollers. The friction between the rollers and the chamber 110 drives the chamber 110 to rotate around the first direction.

[0062] Alternatively, the drive unit 230 includes a drive motor, a drive gear, and a driven gear. The output shaft of the drive motor is connected to the drive gear, and the driven gear is rotatably connected to the frame 210 around a first direction. During cell culture, the cell culture chamber is fixed to the driven gear. The drive gear and the driven gear mesh with each other to drive the driven gear to rotate, and the cell culture chamber can rotate synchronously with the driven gear to achieve cell culture.

[0063] Alternatively, the drive unit 230 includes a drive motor, a driving pulley, and a driven pulley. The output shaft of the drive motor is connected to the driving pulley, and power is transmitted between the driving pulley and the driven pulley via a conveyor belt. The driven pulley is rotatably connected to the frame 210 around a first direction. During cell culture, the cell culture chamber is fixed to the driven pulley, and the cell culture chamber can rotate synchronously with the driven pulley to achieve cell culture.

[0064] The control unit 220 is a feedback loop component widely used in industrial control applications, such as a programmable memory. It typically stores instructions for performing logical operations, sequential control, timing, counting, and arithmetic operations, controlling various types of mechanical equipment or production processes through digital or analog inputs and outputs. In this embodiment, the control unit 220 controls the start, stop, and speed of the drive unit 230. This is something that those skilled in the art can achieve based on the existing functions of the control unit 220; the control principles and methods are not elaborated here.

[0065] refer to Figures 1 to 6 In some embodiments, the drive unit 230 includes a drive member 231 and at least two rotating members 232, with each rotating member 232 arranged sequentially along a second direction. The drive member 231 is connected to the frame 210 and is drively connected to each rotating member 232, driving each rotating member 232 to rotate around its own axis. The axis of each rotating member 232 is parallel to the first direction. The chamber 110 is stacked with adjacent rotating members 232 along a third direction, with the first direction, second direction, and third direction being mutually perpendicular. This enables batch cell culture and improves cell culture efficiency.

[0066] Specifically, the driving component 231 can be a stepper motor, servo motor or other driving motor, and the rotating component 232 can be a cylindrical roller. The driving component 231 is fixedly connected to the frame 210, and each rotating component 232 is rotatably connected to the frame 210 around a first direction. The first direction can be a horizontal front-back direction, the second direction can be a horizontal left-right direction, and the third direction is a vertical up-down direction. The three directions are perpendicular to each other.

[0067] Along the second direction, the rotating components 232 are arranged in parallel sequence, with the same spacing between adjacent rotating components 232. The output shaft of the drive component 231 is connected to each rotating component 232 via belt drive or gear drive. Along the third direction, the cell culture chamber body 110 is placed above any two adjacent rotating components 232. The bottom outer surface of the chamber body 110 is in contact with the outer cylindrical surface of the two rotating components 232, forming a stacked arrangement, i.e., the two rotating components 232 are located below, and the chamber body 110 is located above. The cross-sections of the three are arranged in a triangle. Each rotating component 232 is driven around... In the first direction of rotation, each rotating component 232 drives the chamber 110 to rotate through friction, causing the cell suspension in the culture chamber 111 to form a stable flow field as the chamber 110 rotates. The chamber 110 also drives the carrier 130 to rotate synchronously. When subjected to external impact, if the impact force acts on the chamber 110 in the third direction, the chamber 110 can slide along the outer cylindrical surface of the rotating component 232 in that direction. This sliding alleviates the impact force and avoids rigid collisions between the rotating component 232 and the chamber 110, thus ensuring the structural integrity of the cell culture chamber and the activity of the cells inside. Furthermore, the number of rotating components 232 can be two, three, four, or five. Multiple rotating components 232 arranged in the second direction can simultaneously support multiple cell culture chambers, enabling multiple chambers 110 to conduct cell culture together, thereby improving cell culture efficiency.

[0068] refer to Figures 1 to 6 According to the cell culture process of the embodiments of this application, cell culture is performed using the cell culture device in any of the above embodiments. The cell culture process includes two steps: culture preparation and aggregation growth. In the culture preparation step, a cell suspension is added into the culture chamber 111 until the cell suspension submerges the carrier 130. In the aggregation growth step, the power output of the drive unit 230 is controlled by the control unit 220, causing the drive unit 230 to drive the cell culture chamber to rotate around a first direction. The cells in the cell suspension aggregate on the outer peripheral wall of the carrier 130 until the cells grow to form cell spheres 310. The carrier 130 is provided with an inner cavity 131 and through holes 132, making the carrier 130 a hollow porous structure. Thus, the cells aggregated on the outer peripheral wall of the carrier 130 can be transported with nutrients, oxygen, and cell products through the inner cavity 131 and through holes 132, making the culture microenvironment of the cell spheres 310 more stable and uniform, thereby ensuring the uniformity of the growth size of the cell spheres 310.

[0069] Specifically, the carrier 130 can be made of a hydrophilic material. The cell suspension is immersed in the carrier 130, and the cells in the cell suspension can contact the outer wall of the carrier 130. The carrier 130 is driven by the driving unit 230 to rotate around the first direction. During this process, the cell suspension can also flow around the first direction. The cell suspension comes into contact with the carrier 130, and the cells in the cell suspension aggregate on the outer wall of the carrier 130. As the cell culture chamber continues to rotate, the cells aggregate, grow and differentiate on the outer wall of the carrier 130 until they are formed into cell spheres 310 that at least cover part of the outer wall of the carrier 130.

[0070] During the cultivation of cell spheres 310, nutrients and oxygen can be transported to the central region of cell spheres 310 through the hollow porous space formed by the inner cavity 131 and the through holes 132, ensuring the permeation of cell spheres 310 and enabling them to grow healthily and extend the cultivation cycle. In addition, cell secretion products can also be transported outward through the hollow porous space, facilitating subsequent extraction and application, thereby ensuring the uniformity of the size of cell spheres 310.

[0071] refer to Figures 1 to 6 In some embodiments, during the culture preparation step, a hydrophobic material is coated at intervals on the outer peripheral wall of the carrier 130, so that hydrophobic isolation segments 134 and hydrophilic culture segments 133 are formed on the carrier 130 in sequence along a first direction. During the aggregation growth step, cells aggregate on the outer peripheral wall of the hydrophilic culture segment 133. Thus, by coating with a hydrophobic material, the setting of the hydrophobic isolation segments 134 can be controlled. When the position of the hydrophobic isolation segments 134 needs to be adjusted, the hydrophobic material can be wiped off and recoated, thereby enabling the carrier 130 to be reused and reducing the cost of consumables for cell culture.

[0072] Specifically, the culture preparation step can use a hydrophilic carrier 130 as a base. The carrier 130 extends in a cylindrical shape along a first direction and has an inner cavity 131 extending along the first direction. Multiple radial through-holes 132, communicating with the inner cavity 131, are evenly distributed on the sidewalls. The untreated outer peripheral wall of the carrier 130 can naturally wet the cell suspension. When setting the hydrophobic isolation section 134, hydrophobic materials such as fluorinated resin, superhydrophobic nanoparticles, and phenelzine can be used for spraying. The spraying range can be adjusted according to requirements. Compared to using hydrophobic materials to make the hydrophobic isolation section 134, by spraying hydrophobic material to cover part of the carrier 130, the distribution of the hydrophobic isolation section 134 can be flexibly adjusted according to the required size of the cell spheres 310, which is beneficial to improving the adaptability of the culture process to different culture requirements. Furthermore, the hydrophobic material can be wiped off, restoring the hydrophilic properties of the carrier 130 and allowing for repeated coating and reuse, thus achieving the reuse of consumables.

[0073] It should be noted that the cell suspension is able to be attracted to hydrophilic materials and stay away from hydrophobic materials. During the rotation of the cell culture chamber around the first direction, the hydrophilic culture section 133 can attract the cell suspension, so that the cells in the cell suspension can aggregate on the outer wall of the hydrophilic culture section 133. The hydrophobic isolation section 134 can repel and separate the cell suspension, so as to prevent the cells in the cell suspension from aggregating on the hydrophobic isolation section 134, thereby achieving the isolation of the hydrophilic culture section 133, so as to prevent the cells aggregated on different hydrophilic culture sections 133 from contacting each other, or to prevent the aggregated cells on the hydrophilic culture section 133 from contacting the cover body 120.

[0074] refer to Figures 1 to 6 In some embodiments, the rotational speed of the carrier 130 ranges from 10 r / min to 200 r / min. By limiting the lower limit of the rotational speed, the flow of the cell suspension can be ensured, allowing the cells to aggregate more fully and uniformly, thereby culturing them into cell spheres 310. This also ensures efficient exchange of nutrients, oxygen, and cell products, which is beneficial for extending the culture time of cell spheres 310 and allowing for longer-term observation. Limiting the upper limit of the culture rotational speed helps maintain cell physiological activity, ensures the regularity of cell sphere 310 morphology, and stabilizes material exchange efficiency.

[0075] For example, the rotation speed of cell culture can be 10 r / min, 20 r / min, 30 r / min, 40 r / min, 50 r / min, 60 r / min, 70 r / min, 80 r / min, 90 r / min, 100 r / min, 110 r / min, 120 r / min, 130 r / min, 140 r / min, 150 r / min, 160 r / min, 170 r / min, 180 r / min, 190 r / min or 200 r / min, or within any two of the above values.

[0076] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of this application. Furthermore, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other.

Claims

1. A cell culture pod, comprising: The cell culture cabin comprises: a cabin body provided with a culture cavity; a cover body detachably connected with the cabin body; a carrier arranged along a first direction, the carrier being connected with the cover body, the carrier being located in the culture cavity, the carrier being provided with an inner cavity and a plurality of through holes, the inner cavity being in communication with the culture cavity, each of the through holes penetrating a cavity wall of the inner cavity, each of the through holes being in communication with the inner cavity; the carrier comprises a hydrophilic culture section and a hydrophobic isolation section connected with each other, the hydrophobic isolation section being arranged along the first direction in sequence with the hydrophilic culture section, the hydrophilic culture section being provided with the inner cavity and the through holes, the cover body being connected with the hydrophobic isolation section at an end of the carrier.

2. The cell culture pod of claim 1, wherein, the hydrophobic isolation section is provided with the inner cavity and the through holes.

3. The cell culture pod of claim 1, wherein, a length dimension of the hydrophilic culture section along the first direction is 0.1 mm to 1 mm; and / or, a length dimension of the hydrophobic isolation section along the first direction is 1 mm to 5 mm.

4. The cell culture pod of claim 1, wherein, The cell culture cabin comprises a plurality of the carriers, each of the carriers being connected with the cover body, and the carriers being arranged at intervals.

5. A cell culture device, characterized by, The cell culture cabin comprises: a frame body; a control unit; a driving unit connected with the frame body, and the driving unit being electrically connected with the control unit; the cell culture cabin according to any one of claims 1 to 4, the cabin body being drivingly connected with the driving unit, and the cabin body being configured to be driven to rotate around the first direction by the driving unit.

6. The cell culture device of claim 5, wherein, The driving unit comprises a driving member and at least two rotating members, each of the rotating members being arranged along a second direction in sequence, the driving member being connected with the frame body and drivingly connected with each of the rotating members for driving each of the rotating members to rotate around an axis thereof, the axis of each of the rotating members being parallel to the first direction, the cabin body being arranged in a laminated manner along a third direction with two adjacent rotating members, and the first direction, the second direction and the third direction being perpendicular to each other in pairs.

7. A cell culture process, wherein the cell culture device according to claim 5 or 6 is used for cell culture, characterized in that, The method comprises the following steps: a culture preparation step of adding a cell suspension into the culture cavity until the cell suspension immerses the carrier; a cluster growth step of controlling a power output of the driving unit by the control unit, so that the driving unit drives the cell culture cabin to rotate around the first direction, and cells in the cell suspension are clustered on a peripheral wall of the carrier until the cells grow to form cell spheres.

8. The cell culture process of claim 7, wherein, In the culture preparation step, a hydrophobic material is coated at intervals on the peripheral wall of the carrier, so that the carrier is formed with the hydrophobic isolation section and the hydrophilic culture section connected with each other along the first direction; In the cluster growth step, the cells are clustered on the peripheral wall of the hydrophilic culture section.

9. The cell culture process of claim 7, wherein, A rotation speed of the carrier ranges from 10 r / min to 200 r / min.

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