Apparatus and method for performing cell proliferation and measuring cell proliferation density in real time without contact.

Abstract

The present invention provides an apparatus and method for performing cell proliferation and measuring cell proliferation density in real time without contact, and is used to perform cell proliferation and detect the density of the substance to be measured in a cell culture bag. The apparatus includes a weight sensor mounted on a fixed plate for monitoring weight in real time, a Y-axis oscillation mechanism mounted on the weight sensor for oscillation along the Y-axis, an X-axis oscillation mechanism mounted on the Y-axis oscillation mechanism for oscillation along the X-axis, and a density sensor mounted on the X-axis oscillation mechanism for detecting the density of the substance to be measured in a cell culture bag. The apparatus and method can automate sterile culture of cell culture bags.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims the priority of Chinese Patent Applications No. 2023218446223, No. 2023108631538, No. 202310860536X, No. 202310864234X, No. 2023218491534, No. 2023218446257, and No. 2023218491708, filed on July 14, 2023. The disclosure content thereof is incorporated herein by reference and is equivalent to the content fully described herein.

[0002] The present invention relates to equipment for automated cell culture growth (commonly called a rocking table), and particularly to a device and method for monitoring the supplementary liquid according to the cell growth density and growth multiple online.

Background Art

[0003] GE has disclosed a cell growth device, the structure of which is a sealed 2D rocking table. The disadvantages are that the cell growth device cannot be placed and used in a carbon dioxide culture incubator, and it is necessary to transport carbon dioxide and dissolved oxygen alone inside and extract waste gas, which is relatively complex and prone to contamination. Also, all existing cell growth mixing devices require a single carbon dioxide tube to deeply contact the culture medium liquid inside the cell culture bag for ventilation, which is inconvenient to install, has a high operation requirement level, and strict usage environment requirements, and cannot guarantee aseptic cell culture. Among existing cell growth devices and systems, some adopt a mechanical stirring method, which has the problem that shear force acts on cells, and the cells collide hard with the stirrer and are crushed. Also, because the tube is deeply inserted into the cell culture bag, the rocking mixing angle is small and cannot be completely and uniformly mixed, especially cannot be truly mixed with the medium. As described above, they all rely on manual work and have many disadvantages. The operation equipment for cell culture is not centralized and is not suitable for large - scale cell culture. The manual connection and ventilation operation of the carbon dioxide tube are likely to contaminate the cells and may waste the culture results of several days.

[0004] There are two existing methods for measuring cell density. The first method involves sampling cells by dropping them onto a surface, placing them under a microscope for contact observation, taking a photograph, and then roughly counting the cells within a predetermined area. The second method involves mixing the cells before sampling, performing contact measurement using a flow cytometry device, and then manually counting the cells within that area. This method offers relatively high accuracy. The drawbacks of both methods are that they rely on manual labor and require manual data uploading, making automation impossible. Real-time monitoring of cell proliferation data is not possible, making online data reading and immediate cell density assessment impossible. The equipment is complex to operate, making it unsuitable for large-scale cell cultures, and manual counting is cumbersome and prone to contaminating cells.

[0005] Furthermore, there is only one fluid replacement mode, currently offering manual replacement, scheduled replacement, or quantitative replacement, preventing true automation from being achieved. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] In view of the technical problems raised in the background technology, the present invention aims to solve at least one of the above technical problems, at least in part, by providing an apparatus for performing cell proliferation and measuring cell proliferation density in real time without contact. [Means for solving the problem]

[0007] To achieve the above-mentioned objectives, the present invention provides the following technical solutions: This application relates to a device for performing cell proliferation and measuring cell proliferation density in real time without contact, wherein the device is used to perform cell proliferation and detect the density of the substance to be measured in a cell culture bag. A weight sensor is installed on a fixed plate to monitor the weight in real time, A Y-axis oscillation mechanism is installed on the weight sensor and oscillates along the Y-axis direction, An X-axis rocking mechanism is installed in the Y-axis rocking mechanism and is for rocking along the X-axis direction, It includes a density sensor installed in the X-axis oscillation mechanism for detecting the density of the substance being measured in the cell culture bag.

[0008] In a more preferred embodiment, the cell culture bag is provided with a cell density detection port, and the inside of the cell density detection port is in communication with the space inside the cell culture bag.

[0009] In a more preferred embodiment, the density sensor includes an infrared light source, a spectral prism, a reference infrared receiver A, a detection infrared receiver B, and a signal processing circuit for optical signals.

[0010] In a more preferred embodiment, the infrared light source emits parallel infrared rays in the range of 800 to 1100 nm, the spectral prism is a spectral prism with a division ratio of 1:1, and the parallel infrared rays, after passing through the spectral prism, are divided into a reference parallel ray and a detection parallel ray. The reference parallel ray is detected by a reference infrared receiver A, and the detection parallel ray passes through a cell density detection port and is then detected by a detection infrared receiver B.

[0011] In a more preferred embodiment, the reference parallel ray signal and the absorbed parallel ray signal are entered into an optical signal processing circuit, where the amount of light received from both is compared and corrected, the absorbance value is calculated, and this value is used as an indicator of cell density, thereby indirectly detecting changes in the density value of the object being measured.

[0012] In a more preferred embodiment, the Y-axis oscillating mechanism includes a Y-axis frame, first oscillating shafts located on both sides of the Y-axis frame, and a connecting rod, a first drive rod, and a Y-axis motor that drive at least one of the first oscillating shafts to rotate, wherein support frames are provided on both sides of the weight sensor, and first support shaft holes are provided in the support frames for supporting the first oscillating shafts, the first oscillating shafts are hinged to one end of the connecting rod, and the other end of the connecting rod is hinged to the first drive rod of the Y-axis motor.

[0013] In a more preferred embodiment, the X-axis rocking mechanism includes an X-axis rocking support plate, a second rocking shaft located on one or both sides of the X-axis rocking support plate, and a second drive rod and an X-axis motor that drive the X-axis rocking support plate to rock, wherein the second rocking shaft is provided through a second support shaft hole perpendicular to the first rocking shaft provided in the Y-axis frame, and the second drive rod is hinged to the bottom of the X-axis rocking support plate and is offset from the axis on which the second rocking shaft is located.

[0014] In a more preferred embodiment, the Y-axis frame is formed as a rectangular frame overall, and the X-axis swing support plate is positioned in the center of the Y-axis frame, with a gap around it.

[0015] In a more preferred embodiment, the rotational amplitude and frequency of the Y-axis motor and the X-axis motor are controlled by electrically connecting them to an electrical control board.

[0016] In a more preferred embodiment, a control valve for controlling the connection and disconnection of cell culture bags and culture medium tubes is installed on the X-axis rocking support plate, and the control valve is a peristaltic pump or a clamp valve.

[0017] The present invention further protects a method for performing cell proliferation and measuring cell proliferation density in real time without contact, wherein the method employs an apparatus according to any of the above embodiments and performs the following steps: Step S01 involves placing the entire apparatus inside a carbon dioxide culture incubator, fixing the cell culture bag to the X-axis rocking support plate, fixing the tube of the cell culture bag to the control valve, and fixing the cell density detection port of the cell culture bag to the density sensor. Step S02 involves turning on the power and starting operation, powering the electrical control board, returning the Y-axis motor and X-axis motor to their home positions, adjusting the X-axis swing support plate and Y-axis frame to a perfectly horizontal position, reading the weight with the weight sensor, and completing the preparation of the entire device. Step S03 involves injecting coated cells into a cell culture bag and completing horizontal static culture for a predetermined time. Step S04 involves opening the control valve to inject the culture medium, and then activating the Y-axis motor and / or X-axis motor simultaneously or individually based on the oscillation amplitude and frequency of the Y-axis motor and / or X-axis motor set by the user, thereby gently agitating and mixing the cells and culture medium, and also mixing the carbon dioxide that has permeated the permeable cell culture bag into the cell culture bag. Step S05 involves activating a density sensor to record cell density data in real time, uploading the cell density data to an electrical control board, and then, after the cells have grown for a predetermined number of days, the cells proliferate in large numbers. A method comprising step S06, which involves reading data from a weight sensor after the culture is complete to obtain the accurate cell weight.

[0018] In a more preferred embodiment, in step S05, the control valve is opened again according to the user's needs or program settings, the same or a different culture medium is injected, and the mixing of the culture medium and cells is continued again according to a new density setting or a predetermined oscillation amplitude and frequency, so that the cells are mixed with the culture medium and carbon dioxide, allowing the cells to absorb nutrients and grow and proliferate. [Effects of the Invention]

[0019] This invention overcomes the shortcomings of existing technologies and provides an automated cell production apparatus that thoroughly mixes cells by selecting appropriate oscillation angles, amplitudes, and frequency levels according to the cell type. The apparatus enables automated sterile culture of cell culture bags, and no human intervention is required throughout the entire process during the culture period. The cell proliferation system apparatus integrates a density sensor that can monitor the cell proliferation ratio in real time, and provides a real-time fluid replenishment function according to the cell proliferation ratio, providing true fluid replenishment (such as culture medium) according to the cell proliferation ratio and cell state, as well as providing multiple configurable replenishment modes. [Brief explanation of the drawing]

[0020] [Figure 1] It is a schematic structural diagram of a cell proliferation device according to an embodiment of the present invention. [Figure 2] It is a schematic structural diagram of the other side of a cell proliferation device according to an embodiment of the present invention. [Figure 3] It is a schematic structural diagram of a density sensor and a cell culture bag according to an embodiment of the present invention. [Figure 4] It is a schematic diagram of the detection principle of a density sensor according to an embodiment of the present invention. [Figure 5] It is a statistical diagram of cell growth data using a cell proliferation device according to an embodiment of the present invention. [Figure 6] It is a flowchart of cell proliferation culture and detection steps according to an embodiment of the present invention.

Modes for Carrying Out the Invention

[0021] Hereinafter, the features and exemplary embodiments of various aspects of the present invention will be described in detail. In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the drawings and embodiments. It should be understood that the specific embodiments described herein are only configured to interpret the patent of the present invention and are not configured to limit the patent of the present invention. For those skilled in the art, the present invention can be implemented even if some of these specific details are omitted. The following description of the embodiments is for the purpose of providing a better understanding of the present invention by showing examples of the present invention.

[0022] In this specification, the terms "1," "2," etc., are merely for distinguishing one entity or operation from another, and do not necessarily require or imply that there is an actual relationship or order between these entities or operations. Furthermore, the terms "includes," "contains," or any variation thereof are intended to be non-exclusive, so that a process, method, article, or apparatus containing a set of elements includes not only those elements but also other elements not explicitly listed, or further elements specific to such process, method, article, or apparatus. Unless further limited, the elements limited by the statement "includes" do not preclude the existence of other identical elements within a process, method, article, or apparatus containing such elements.

[0023] Referring to Figures 1 and 2, the cell proliferation apparatus of the present invention includes a fixed plate 1, a weight sensor 9, a Y-axis frame 2, a Y-axis motor 7, an X-axis oscillating support plate 5, an X-axis motor 10, a cell culture bag 4, a density sensor 3, and an electrical control board 8. The entire cell proliferation apparatus can be placed inside a carbon dioxide culture incubator.

[0024] Here, the weight sensor 9 is mounted on the fixed plate 1, support frames 12 are installed on both sides of the weight sensor 9, a first support shaft hole 121 is installed near the top of the support frame 12, the Y-axis frame 2 as a whole has a rectangular shape, a first pivot shaft 21 is installed at the center outside the two long sides of the Y-axis frame, the first pivot shaft 21 passes through the first support shaft hole 121, and the fitting of the first pivot shaft 21 and the first support shaft hole 121 allows the Y-axis frame 2 to rotate / oscillate relative to the support frame 12. Furthermore, one of the first pivot shafts 21 is hinged to one end of the connecting rod 22, and the other end of the connecting rod 22 is hinged to the first drive rod 71 of the Y-axis motor 7. The Y-axis motor 7 is electrically connected to an electrical control board 8. The Y-axis motor 7 drives the extension and retraction of the first drive rod 71 and also drives the oscillation of the connecting rod 22. The oscillation of the connecting rod 22 is converted into rotation of the first pivot shaft 21, thereby enabling the Y-axis frame 2 to oscillate from side to side. The rotational amplitude and frequency of the Y-axis motor 7 can be controlled via the control board 8, thereby controlling the oscillation amplitude and frequency of the Y-axis frame 2.

[0025] The X-axis oscillation support plate 5 is formed in a plate shape and is similar in shape to the Y-axis frame 2, which is generally a rectangular frame, but its overall size is slightly smaller than that of the Y-axis frame 2. The X-axis oscillation support plate 5 is positioned in the center of the rectangular frame of the Y-axis frame 2, and a gap is left around it between the X-axis oscillation support plate 5 and the Y-axis frame 2 to ensure that the X-axis oscillation support plate 5 can oscillate within the Y-axis frame. Second oscillation shafts 51 are installed in the center of the short sides of both sides of the X-axis oscillation support plate 5, and second support shaft holes 23 are installed at positions corresponding to the second oscillation shafts 51 on both short sides of the Y-axis frame 2. The second oscillation shafts 51 on both sides of the X-axis oscillation support plate 5 are inserted into the second support shaft holes 23 in the center of the short sides of both short sides of the Y-axis frame 2, and the fitting of the second oscillation shafts 51 and the second support shaft holes 23 causes the X-axis oscillation support plate 5 to rotate / oscillate relative to the Y-axis frame 2. Preferably, the second oscillation shafts 51 and the second support shaft holes 23 are installed on only one side. More preferably, a U-shaped lateral support plate 122 is extended downward on both sides of the Y-axis frame 2 (i.e., at the position of the first pivot axis 21), and the X-axis motor 10 is installed at a position on one side of the lateral plate of the U-shaped lateral support plate 122 that is close to the support frame 12, i.e., the X-axis motor 10 is at a predetermined distance from the center of the U-shaped lateral support plate 122. The output end of the X-axis motor 10 is connected to a second drive rod 101, and the other end of the second drive rod 101 is hinged to the X-axis oscillating support plate 5. The hinge connection position is offset by a predetermined distance from the axes of the second oscillating shafts 51 on both sides. As a result, the X-axis motor 10 drives the extension and retraction of the second drive rod 101, thereby allowing the X-axis oscillating support plate 5 to oscillate back and forth around the second oscillating shafts 51. Preferably, the X-axis motor 10 is electrically connected to an electrical control board 8, and the rotational amplitude and frequency of the X-axis motor 10 can be controlled through the control board 8, thereby controlling the oscillation amplitude and frequency of the X-axis oscillating support plate 5.

[0026] By employing the cell proliferation apparatus described above in the present invention, the following beneficial technical effects can be obtained. 1. It meets the need to culture coated cells horizontally for multiple days and then the need to mix the cells after they have proliferated in large quantities, providing multiple mixing modes and angles to achieve differentiated cell production. It can provide a shear-free mixing mode using vortex oscillation, a cell aggregate-free mixing mode using wave oscillation, and a small-amplitude mixing mode. Cells can be completely fixed to the surface of the oscillation support plate. Another sensor can be integrated to enable monitoring during the cell culture process.

[0027] 2. The entire rocking platform is the key component for achieving fully automated cell production, possessing the highest level of automation and best integration.

[0028] 3. It provides key technologies required in the process from coating to culturing various cells, such as vibrational detachment of primary adherent or semi-suspended cells, flow utilizing gravity gradient of primary cells, horizontal mode by oscillating mixing during culture, shear mode by vortex flow, and aggregate dispersion mode by wave. It also monitors the total volume of cell culture bags in real time, provides weight data for cell collection at the end of culture, and performs weighing after the completion of culture.

[0029] 4. Throughout the entire culture process, the agitation mixing mode is highly customizable, and multiple parameters are adjustable. This makes it easier to classify various cell types and formulate individual culture plans during the culture process, significantly improving the culture efficiency of various cell types by summarizing culture experience over multiple cycles. The control system can control the cell culture flow in real time and can be used for remote control. Cameras can be installed to monitor cell morphology in real time.

[0030] The following is data on cell mixing and growth under various rocking modes of the rocking platform.

[0031] [Table 1]

[0032] Furthermore, referring to Figure 3, the cell culture bag 4 is equipped with a cell density detection port 41, which is integrated into the edge of the cell culture bag 4 by welding. The inside of the cell density detection port 41 communicates with the space inside the cell culture bag 4, and when the rocking platform rocks, the liquid inside the cell culture bag 4 can flow into the cell density detection port 41. The density sensor 3 is installed on the X-axis rocking support plate 5, and when the cell culture bag 4 is fixed to the X-axis rocking support plate 5, the cell density detection port 41 can be inserted into the density sensor 3. Preferably, the density sensor 3 is installed on one corner of the X-axis rocking support plate 5.

[0033] Furthermore, a control valve 6 is installed on the X-axis oscillation support plate 5, preferably a peristaltic pump or a clamp valve, which secures the tube of the cell culture bag 4 to the control valve 6.

[0034] The optical principle, structure, and circuit principle of the density sensor 3 are shown in Figure 4.

[0035] By employing an infrared light source in the 1,800-1,100 nm range, it provides a stable and reliable parallel light source. Light in this wavelength range has an extremely high absorption rate for the object being measured, making it convenient for detecting the infrared absorption rate of the object being measured.

[0036] 2. Parallel rays from an infrared light source pass through a spectral prism with a division ratio of 1:1, splitting the light source into two, which are provided as a reference light source and a detection light source, respectively, and used for detection and light source intensity correction.

[0037] 3. Reference infrared receiver A monitors the stability of the light source in real time and provides a basis for correction to calculate subsequent changes in incident light intensity values.

[0038] 4. The cell density detection port 41 is integrated into the edge of the cell culture bag 4 by welding, and the inside of the cell density detection port 41 communicates with the space inside the cell culture bag 4, so that when the rocking platform rocks, the liquid inside the cell culture bag 4 can flow into the inside of the cell density detection port 41. Preferably, the detection area of ​​the cell density detection port 41 is highly transparent and can allow light to pass through.

[0039] 5.1:1 spectral prism detected parallel light rays pass through cell density detection port 41, and the parallel light after absorption by the object being measured enters detection infrared receiver B.

[0040] 6. The reference parallel ray signal received by reference infrared receiver A and the absorbed parallel ray signal received by detection infrared receiver B are input. The received light amounts obtained from reference infrared receiver A and detection infrared receiver B are compared and corrected to calculate the incident light intensity and received light intensity at this point. Furthermore, the absorbance value is calculated and used as an indicator of cell density, thereby indirectly detecting changes in the density value of the object being measured.

[0041] By employing the density sensor of the present invention, the following beneficial technical effects can be obtained: 1) The entire sensor is completely sealed and does not come into contact with the outside world, thus avoiding the risk of light or circuitry being affected by the external environment, and it can be used repeatedly.

[0042] 2) The cell density detection port is completely independently integrated into the cell culture bag of the cell culture system, undergoes high-temperature sterilization and other processes, and is disposable with no risk of contamination.

[0043] 3) Data from the entire cell culture process is recorded in real time, making it easy to read the data and obtain changes in the curve to determine the extent of cell proliferation and growth status. This allows operators to understand the situation in real time and conveniently perform corresponding operations such as adding nutrients or ending the culture.

[0044] 4) Cell growth density is obtained as intuitive feedback, which is useful for summarizing the growth status of various cells in the culture process, classifying them by cell type, and formulating individual culture plans, thereby significantly increasing cell culture efficiency.

[0045] Figure 5 shows data from various cells grown for 14 days, and it can be seen from the figure that the linearity is very high.

[0046] The following describes a method for cell proliferation culture and detection using the cell proliferation apparatus and density sensor of the present invention, and the specific steps include the following. S01: The entire cell proliferation apparatus is placed inside the carbon dioxide culture incubator together with the fixing plate 1, the cell culture bag 4 is fixed to the X-axis rocking support plate 5, the tube of the cell culture bag 4 is fixed to the peristaltic pump / clamp valve 6, and the cell density detection port 41 of the cell culture bag 4 is fixed to the density sensor 3.

[0047] S02: When the power is turned on and operation begins, power is supplied to the electrical control board 8, the Y-axis motor 7 and X-axis motor 10 return to their home positions, that is, the X-axis swing support plate and Y-axis frame 2 return to a completely horizontal position, the weight sensor 9 reads the weight, and the entire device is ready.

[0048] S03: When culturing cells, the coated cells are injected into cell culture bag 4 and the horizontal static culture is completed for a predetermined time.

[0049] S04: After horizontal static culture, the peristaltic pump / clamp valve 6 can be opened to inject the culture medium, and the Y-axis motor 7 and / or X-axis motor 10 can be started simultaneously or individually based on the oscillation amplitude and angle set by the user, generating oscillation in one or two directions on the X-axis oscillation support plate 5, thereby gently agitating and mixing the cells and culture medium, and mixing the carbon dioxide that has permeated the permeable cell culture bag 4 into the cell culture bag.

[0050] S05: The density sensor 3 is activated to record cell density data in real time and upload it to the electrical control board 8. After the cells have grown for a predetermined number of days, the cells proliferate in large numbers. Depending on the user's needs / program settings, the peristaltic pump / clamp valve 6 is opened again, the same or a different culture medium is injected, and the mixing of the culture medium and cells continues according to the new density setting / predetermined oscillation amplitude and angle (or maintain the previous oscillation amplitude and angle), and frequency, so that the cells are thoroughly mixed with the culture medium and carbon dioxide, allowing them to absorb nutrients and grow and proliferate.

[0051] S06: After the culture is complete, read the data from weight sensor 9 to obtain the accurate cell weight.

[0052] Furthermore, as shown in Figure 6, this is a specific example in which coated NK cells are used as an example and cell proliferation culture is performed from day 5 to day 11 using the apparatus and method of the present invention.

[0053] Clearly, those skilled in the art can make various changes and modifications to the embodiments of the present invention without departing from the spirit and scope of the invention. Therefore, if these changes and modifications fall within the scope of the claims of the present invention and the equivalent art, the present invention is intended to include these changes and modifications as well. [Explanation of Symbols]

[0054] 1: Fixed plate 2: Y-axis frame 21:First swing axis 22: Connecting rod 23:Second support shaft hole 3: Density sensor 4: Cell culture bag 41: Cell density detection port 5: X-axis oscillating support plate 51:Second swing axis 6: Peristaltic pump / clamp valve 7: Y-axis motor 71: First drive rod 8: Electrical control board 9: Weight sensor 10: X-axis motor 101: Second drive rod 12: Support frame 121:First support shaft hole 122: U-shaped lateral support plate.

Claims

1. A device for performing cell proliferation and measuring cell proliferation density in real time without contact, the device being used to perform cell proliferation and detect the density of the substance to be measured in a cell culture bag, A weight sensor is installed on a fixed plate to monitor the weight in real time, A Y-axis oscillation mechanism is installed on the weight sensor and oscillates along the Y-axis direction, An X-axis oscillating mechanism is installed in the Y-axis oscillating mechanism and oscillates along the X-axis direction, An apparatus comprising: a density sensor mounted on an X-axis oscillating mechanism for detecting the density of a substance to be measured in a cell culture bag;

2. The apparatus according to claim 1, characterized in that the cell culture bag is provided with a cell density detection port, and the inside of the cell density detection port is in communication with the space inside the cell culture bag.

3. The apparatus according to claim 2, characterized in that the density sensor includes an infrared light source, a spectral prism, a reference infrared receiver A, a detection infrared receiver B, and a signal processing circuit for optical signals.

4. The apparatus according to claim 3, characterized in that the infrared light source emits parallel infrared rays in the range of 800 to 1100 nm, the spectral prism is a spectral prism with a division ratio of 1:1, the parallel infrared rays are separated into a reference parallel ray and a detection parallel ray after passing through the spectral prism, the reference parallel ray is detected by a reference infrared receiver A, and the detection parallel ray is detected by a detection infrared receiver B after passing through a cell density detection port.

5. The apparatus according to claim 4, characterized in that the reference parallel ray signal and the absorbed parallel ray signal are entered into an optical signal processing circuit, the amount of light received by both is compared and corrected, the absorbance value is calculated and used as an index representing the density of cells, thereby indirectly detecting a change in the density value of the object being measured.

6. The apparatus according to claim 1, wherein the Y-axis oscillating mechanism includes a Y-axis frame, first oscillating shafts located on both sides of the Y-axis frame, and a connecting rod, a first drive rod, and a Y-axis motor that drive at least one of the first oscillating shafts to rotate, and support frames are installed on both sides of the weight sensor, and first support shaft holes for supporting the first oscillating shafts are provided in the support frames, the first oscillating shafts are hinged to one end of the connecting rod, and the other end of the connecting rod is hinged to the first drive rod of the Y-axis motor.

7. The X-axis oscillation mechanism includes an X-axis oscillation support plate, a second oscillation shaft located on one or both sides of the X-axis oscillation support plate, and a second drive rod and an X-axis motor that drive the X-axis oscillation support plate to oscillate, wherein the second oscillation shaft is provided through a second support shaft hole perpendicular to the first oscillation shaft provided in the Y-axis frame, and the second drive rod is hinged to the bottom of the X-axis oscillation support plate and offset from the axis on which the second oscillation shaft is located, as described in claim 6.

8. The apparatus according to claim 7, characterized in that the Y-axis frame is formed as a rectangular frame overall, and the X-axis oscillation support plate is positioned in the center of the Y-axis frame and has a gap around it.

9. The apparatus according to claim 7 or 8, characterized in that the rotational amplitude and frequency of the Y-axis motor and the X-axis motor are controlled by electrically connecting them to an electrical control board.

10. The apparatus according to claim 7, wherein a control valve for controlling the connection and disconnection of cell culture bags and culture medium tubes is installed on the X-axis oscillating support plate, and the control valve is a peristaltic pump or a clamp valve.

11. A method for performing cell proliferation and measuring cell proliferation density in real time without contact, wherein the method employs the apparatus described in any one of claims 1 to 10. Step S01 involves placing the entire apparatus inside a carbon dioxide culture incubator, fixing the cell culture bag to the X-axis rocking support plate, fixing the tube of the cell culture bag to the control valve, and fixing the cell density detection port of the cell culture bag to the density sensor. Step S02 involves turning on the power and starting operation, powering the electrical control board, returning the Y-axis motor and X-axis motor to their home positions, adjusting the X-axis swing support plate and Y-axis frame to a completely horizontal position, reading the weight with the weight sensor, and completing the preparation of the entire device. Step S03 involves injecting coated cells into a cell culture bag and completing horizontal static culture for a predetermined time. Step S04 involves opening the control valve to inject the culture medium, and then activating the Y-axis motor and / or X-axis motor simultaneously or individually based on the oscillation amplitude and frequency of the Y-axis motor and / or X-axis motor set by the user, thereby gently agitating and mixing the cells and culture medium, and also mixing the carbon dioxide that has permeated the permeable cell culture bag into the cell culture bag. Step S05 involves activating a density sensor to record cell density data in real time, uploading the cell density data to an electrical control board, and after the cells have grown for a predetermined number of days, the cells proliferate in large numbers. A method comprising step S06, after the completion of culture, reading data from a weight sensor to obtain the accurate cell weight.

12. The method according to 11, characterized in that in step S05, the control valve is opened again according to the user's needs or program settings, the same or a different culture medium is injected, and the mixing of the culture medium and cells is continued again according to a new density setting or a predetermined oscillation amplitude and frequency, so that the cells are mixed with the culture medium and carbon dioxide so that the cells can absorb nutrients and grow and proliferate.

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