CN220597520U - Device for cell expansion and non-contact real-time measurement of cell expansion density - Google Patents

Abstract

The utility model provides a device for cell expansion and non-contact real-time measurement of cell expansion density, which is used for detecting the density of an object to be detected in a cell culture bag, and comprises the following components: the weighing sensor is arranged on the fixed plate and used for monitoring weight in real time; the Y-axis swinging mechanism is arranged on the weighing sensor and is used for swinging along the Y-axis direction; the X-axis swinging mechanism is arranged on the Y-axis swinging mechanism and is used for swinging along the X-axis direction; the density sensor is arranged on the X-axis swinging mechanism and used for detecting the density of an object to be detected in the cell culture bag. The device can realize the automatic sterile culture of the cell culture bag, and no manual participation is needed in the whole process.

Description

Device for cell expansion and non-contact real-time measurement of cell expansion density

Technical Field

The utility model relates to a device (commonly called a swing table) for automatic cell culture expansion, in particular to a device for monitoring cell expansion density on line and supplementing liquid according to expansion times.

Background

The GE company discloses a cell expander which is a closed 2D swing table, has the defects that the cell expander cannot be placed in a carbon dioxide incubator for use, carbon dioxide and dissolved oxygen are required to be independently conveyed to the inside, waste gas is pumped out, and the cell expander is relatively complex and easy to dye bacteria. In addition, the existing cell expansion and mixing device needs to be provided with a separate carbon dioxide pipeline to deeply contact with the culture solution in the cell culture bag for ventilation, is inconvenient to install, has high operation requirement and high use environment requirement, and cannot ensure sterile culture of cells; the existing cell expansion device or system adopts a mechanical stirring mode, has shearing force on cells, causes hard collision and hard cell breakage of the cells and a stirrer, has small swinging and uniform mixing angle because of the fact that a pipeline goes deep into a cell culture bag, and cannot be truly and uniformly mixed, especially cannot be fully and uniformly mixed with a culture medium; as described above, all rely on manual operation and have a number of disadvantages. The operation equipment for cell culture is not centralized, is not suitable for large-batch cell culture, and is easy to pollute cells due to ventilation of the artificial carbon dioxide gas insertion tube, so that the culture results are scrapped for a plurality of days.

There are two existing cell density measurement modes: firstly, sampling the cell by dripping, placing the cell into a microscope for contact observation, photographing, and then roughly counting the cell with a certain area; the second is to mix and sample the cells uniformly, and to use a flow cytometer to make contact measurement, count after manual circling, and be more accurate. Disadvantages: both the two modes rely on manual operation, and data is uploaded manually, so that automation cannot be realized. The cell growth data cannot be monitored in real time, the data cannot be read on line, and the cell density can be obtained in time. The operation equipment is complex, and is not suitable for large-batch cell culture, the manual counting is complicated, and the cells are easy to pollute.

And the liquid supplementing mode is single, so that manual liquid supplementing, timing or quantitative liquid supplementing cannot be realized at present, and real automation is not realized.

Disclosure of Invention

In view of the technical problems set forth in the background art, the present utility model provides a device for cell expansion and non-contact real-time measurement of cell expansion density, so as to at least partially solve at least one of the technical problems.

In order to achieve the above purpose, the present utility model provides the following technical solutions:

the utility model relates to a device for cell expansion and non-contact real-time measurement of cell expansion density, which is used for cell expansion and detection of the density of an object to be detected in a cell culture bag, and comprises the following components:

the weighing sensor is arranged on the fixed plate and used for monitoring weight in real time;

the Y-axis swinging mechanism is arranged on the weighing sensor and is used for swinging along the Y-axis direction;

the X-axis swinging mechanism is arranged on the Y-axis swinging mechanism and is used for swinging along the X-axis direction;

the density sensor is arranged on the X-axis swinging mechanism and used for detecting the density of an object to be detected in the cell culture bag.

In a further preferred embodiment, the cell culture bag is provided with a cell density detection port, and the inside of the cell density detection port communicates with the space in the cell culture bag.

In a further preferred embodiment, the density sensor comprises an infrared light source, a beam splitting prism, a reference infrared receiver A, a detection infrared receiver B and an optical signal processing circuit.

In a further preferred embodiment, the infrared light source emits 800-1100nm infrared parallel rays, and the beam splitting prism is 1: the device comprises a 1 beam splitting prism, wherein infrared parallel light rays are divided into reference parallel light rays and detection parallel light rays after passing through the beam splitting prism, the reference parallel light rays are detected by a reference infrared receiver A, and the detection parallel light rays pass through a cell density detection port and are detected by a detection infrared receiver B.

In a further preferred embodiment, the reference parallel light signal and the absorbed parallel light signal enter an optical signal processing circuit, the illumination values of the reference parallel light signal and the absorbed parallel light signal are compared and corrected, the light absorbance value is calculated, the density of the cell is represented, and therefore the change of the density value of the measured object is indirectly detected.

Further preferred embodiment, the Y-axis swinging mechanism comprises a Y-axis support, first swinging shafts positioned on two sides of the Y-axis support, a connecting rod for driving at least one side of the first swinging shafts to rotate, a first driving rod and a Y-axis motor, wherein supporting frames are arranged on two sides of the weighing sensor, first supporting shaft holes for supporting the first swinging shafts are formed in the supporting frames, the first swinging shafts are hinged to one ends of the connecting rod, and the other ends of the connecting rod are hinged to the first driving rod of the Y-axis motor.

In a further preferred embodiment, the X-axis swinging mechanism includes an X-axis swinging support plate, a second swinging shaft located at one side or two sides of the X-axis swinging support plate, and a second driving rod and an X-axis motor for driving the X-axis swinging support plate to swing, where the second swinging shaft is inserted into a second supporting shaft hole perpendicular to the first swinging shaft on the Y-axis bracket, and the second driving rod is hinged at the bottom of the X-axis swinging support plate and deviates from the axis where the second swinging shaft is located.

In a further preferred embodiment, the Y-axis support is a rectangular frame, and the X-axis swing supporting plate is disposed in the middle of the Y-axis support and has gaps around the Y-axis support.

In a further preferred embodiment, the Y-axis motor and the X-axis motor are electrically connected to an electronic control board to control the rotational amplitude and frequency of the Y-axis motor and the X-axis motor.

In a further preferred embodiment, the X-axis swing supporting plate is provided with a control valve, the control valve is used for controlling the on-off of the cell culture bag and the culture medium liquid pipeline, and the control valve is a peristaltic pump or a pinch valve.

The utility model overcomes the defects in the prior art, and provides a device for automatically producing cells, which selects corresponding swing angles, amplitude and frequency grades according to cell types and fully mixes the cells; the device can realize the automatic sterile culture of the cell culture bag, and no manual participation is needed in the whole process. The cell expansion system device integrates a density sensor capable of monitoring the cell expansion times in real time, provides the function of supplementing liquid in real time according to the cell expansion times, truly realizes the liquid supplementing according to the cell expansion times or according to the cell state (such as a culture medium), and provides various settable liquid supplementing modes.

Drawings

FIG. 1 is a schematic diagram showing a cell expansion apparatus according to an embodiment of the present utility model;

FIG. 2 is a schematic diagram showing another embodiment of a cell expansion apparatus according to the present utility model;

FIG. 3 is a schematic diagram of the structure of a density sensor and cell culture bag according to an embodiment of the utility model;

FIG. 4 is a schematic diagram of a density sensor detection principle according to an embodiment of the present utility model;

FIG. 5 is a statistical chart of data showing cell growth using the cell expansion device of the present utility model;

FIG. 6 is a flow chart of the cell expansion culture and detection steps according to one embodiment of the present utility model.

In the figure: the device comprises a 1-fixed plate, a 2-Y-axis bracket, a 21-first swinging shaft, a 22-connecting rod, a 23-second supporting shaft hole, a 3-density sensor, a 4-cell culture bag and a 41-cell density detection port; the device comprises a 5-X axis swing supporting plate, a 51-second swing shaft, a 6-peristaltic pump/pinch valve, a 7-Y axis motor, a 71-first driving rod, an 8-electric control plate, a 9-weighing sensor, a 10-X axis motor, a 101-second driving rod, a 12-supporting frame, a 121-first supporting shaft hole and a 122-U-shaped transverse supporting plate.

Detailed Description

Features and exemplary embodiments of various aspects of the present utility model are described in detail below, and in order to make the objectives, technical solutions, and advantages of the present utility model more apparent, the present utility model is described in further detail below with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are merely configured to explain the present patent and are not configured to limit the present patent. It will be apparent to one skilled in the art that the present utility model may be practiced without some of these specific details. The following description of the embodiments is merely intended to provide a better understanding of the present utility model by showing examples of the present utility model.

It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising … …" does not exclude the presence of other like elements in a process, method, article or apparatus that comprises the element.

Referring to fig. 1 and 2, the cell expansion device comprises a fixed plate 1, a weighing sensor 9, a Y-axis bracket 2, a Y-axis motor 7, an X-axis swinging support plate 5, an X-axis motor 10, a cell culture bag 4, a density sensor 3 and an electric control plate 8. The whole cell expansion device can be placed in a carbon dioxide incubator.

The weighing sensor 9 is arranged on the fixed plate 1, supporting frames 12 are arranged on two sides of the weighing sensor 9, a first supporting shaft hole 121 is formed in a position, close to the top, of the supporting frames 12, the whole Y-axis support 2 is a rectangular frame, a first swinging shaft 21 is arranged in the middle position of the outer sides of two long sides of the Y-axis support 2, the first swinging shaft 21 penetrates through the first supporting shaft hole 121, and the Y-axis support 2 can rotate/swing relative to the supporting frames 12 by means of the cooperation of the first swinging shaft 21 and the first supporting shaft hole 121. Further, the first swinging shaft 21 at one side is hinged with one end of a connecting rod 22, the other end of the connecting rod 22 is hinged with a first driving rod 71 of a Y-axis motor 7, the Y-axis motor 7 is electrically connected with an electric control board 8, the Y-axis motor 7 drives the first driving rod 71 to do telescopic motion and drives the connecting rod 22 to swing, the swinging of the connecting rod 22 is converted into the rotation of the first swinging shaft 21, the left-right swinging of the Y-axis bracket 2 is further realized, the rotation amplitude and frequency of the Y-axis motor 7 can be controlled through a control board 8, and the swinging amplitude and frequency of the Y-axis bracket 2 are further controlled.

The X-axis swinging support plate 5 is plate-shaped and basically similar to the Y-axis support 2 of the rectangular frame in shape, but the whole size is slightly smaller than that of the Y-axis support 2, the X-axis swinging support plate 5 is arranged in the middle of the rectangular frame of the Y-axis support 2, and gaps are reserved between the X-axis swinging support plate 5 and the Y-axis support 2 at the periphery, so that the X-axis swinging support plate 5 can swing in the Y-axis support. The second swinging shafts 51 are arranged in the middle of the short sides of the two sides of the X-axis swinging support plate 5, the second supporting shaft holes 23 are arranged at positions corresponding to the two short sides of the Y-axis support frame 2, the second swinging shafts 51 at the two sides of the X-axis swinging support plate 5 respectively penetrate into the second supporting shaft holes 23 in the middle of the two short sides of the Y-axis support frame 2, the X-axis swinging support plate 5 also rotates/swings relative to the Y-axis support frame 2 by the cooperation of the second swinging shafts 51 and the second supporting shaft holes 23, and preferably, the second swinging shafts 51 and the second supporting shaft holes 23 are arranged on one side only. It is further preferred that a U-shaped transverse support plate 122 is provided to extend downward on both sides of the Y-axis bracket 2 (i.e., at the position of the first swing axis 21), and the X-axis motor 10 is provided on the transverse plate of the U-shaped transverse support plate 122 at a position close to the one-side support frame 12, that is, the X-axis motor 10 is located at a distance from the center of the transverse support plate 122. The output end of the X-axis motor 10 is connected with the second driving rod 101, the other end of the second driving rod 101 is hinged to the X-axis swinging support plate 5, the hinged position deviates from the axes of the second swinging shafts 51 at two sides by a certain distance, so that the second driving rod 101 is pushed to stretch and retract under the driving of the X-axis motor 10, the X-axis swinging support plate 5 can swing around the second swinging shafts 51, preferably, the X-axis motor 10 is electrically connected with the electric control plate 8, and the rotation amplitude and frequency of the X-axis motor 10 can be controlled through the control plate 8, so that the swinging amplitude and frequency of the X-axis swinging support plate 5 are controlled.

The cell expansion device disclosed by the utility model has the following beneficial technical effects:

1. the coated cells are subjected to horizontal stationary culture for a plurality of days; the method is characterized in that a plurality of uniformly-mixed modes and angles are provided for the cells after the expansion, so as to realize differentiated production cells; the vortex swing shearing-force-free mixing mode can be provided, the wave swing mixing mode for realizing the aggregation-free cell mass can be provided, and the small-amplitude mixing mode can be provided; the cells can be completely fixed on the table surface of the swinging supporting plate; can integrate other sensors and realize monitoring the cell culture process.

2. The whole swing table is a key part for realizing fully-automatic production of cells, and has the highest automation degree and highest integration.

3. Providing a plurality of key technologies needed in the process of coating cells to culture, such as primary adherence or vibration detachment of semi-suspension cells, wherein the primary cells flow obliquely by gravity; a plurality of independent preset modes such as a horizontal mode, an eddy current non-shearing mode, a wave scattering and agglomerating mode and the like which are uniformly mixed in a swinging way in culture; the total amount of the cell culture bag is monitored in real time, the cells are collected for the completion of the culture, weight data are provided, and weighing and metering are performed after the completion of the culture.

4. In the whole culture process, the swing mixed mode is highly customized, a plurality of parameters are adjustable, different culture plans are formulated for different cells according to categories in the convenient culture process, culture experience is summarized for a plurality of times, and the culture efficiency of different cells is greatly improved. The control system controls the flow of cell culture in real time, can be used for remote control, and can be used for expanding and installing a camera to monitor the cell morphology in real time.

The following is data for cell mixing and growth under various swing modes of the swing table.

Further, referring to fig. 3, a cell density detecting port 41 is provided in the cell culture bag 4, the cell density detecting port 41 is welded to the edge of the cell culture bag 4, the inside of the cell density detecting port 41 communicates with the space in the cell culture bag 4, and when the swing table swings, the liquid in the cell culture bag 4 can flow into the inside of the cell density detecting port 41. The density sensor 3 is provided on the X-axis swing pallet 5, and the cell density detecting port 41 can be inserted into the density sensor 3 when the cell culture bag 4 is fixed to the X-axis swing pallet 5. Preferably, the density sensor 3 is provided at one corner of the X-axis swing pallet 5.

Further, a control valve 6 is further provided on the X-axis swing support plate 5, and preferably, the control valve 6 is a peristaltic pump or a pinch valve, and the tube of the cell culture bag 4 is fixed on the control valve 6.

The optical principle, structure and circuit principle of the density sensor 3 are shown in fig. 4,

1. an infrared light source of 800-1100nm is adopted to provide a stable and reliable parallel light source, and the light of the wave band has extremely high absorptivity to the measured object, so that the absorptivity to the infrared light of the measured object can be conveniently detected.

2. Parallel rays of the infrared light source pass through the 1:1 beam splitter prism to divide the light source into two parts, and the two parts are respectively provided as a reference light source and a detection light source for detection and light source intensity correction.

3. And the reference infrared receiver A is used for monitoring the stability of the light source in real time and providing a correction basis for the subsequent calculation of the change of the incident light intensity value.

4. The cell density detecting port 41 is welded and integrated to the edge of the cell culture bag 4, and the inside of the cell density detecting port 41 is communicated with the space inside the cell culture bag 4, so that when the swing table swings, the liquid inside the cell culture bag 4 can flow into the inside of the cell density detecting port 41. Preferably, the cell density detection port 41 is highly transparent at the detection site and can transmit light.

5. Through 1: the parallel light after the 1 spectroscope passes through the cell density detection port 41, and the parallel light absorbed by the detected object enters the detection infrared receiver B.

6. The reference parallel light signal received by the reference infrared receiver A and the absorbed parallel light signal received by the detection infrared receiver B enter, the illumination values obtained by the reference infrared receiver A and the detection infrared receiver B are compared and corrected, the incident light intensity and the received light intensity at the moment are calculated, the absorbance value is calculated, the density of cells is represented, and the density value change of an object to be detected is indirectly detected.

The density sensor of the utility model has the following beneficial technical effects:

1) The whole sensor is completely closed, does not have any contact with the outside, avoids the risk that light and a circuit are influenced by the outside, and can be repeatedly used.

2) The cell density detection port is completely and independently integrated in a cell culture bag of a cell culture system, and is disposable after being subjected to treatment such as high-temperature sterilization and the like without pollution risk.

3) The whole cell culture process data is recorded in real time, so that the data can be conveniently read, curve change is obtained, the amplitude and growth condition of cell proliferation are obtained, and an operator can conveniently know the condition in real time so as to perform targeted operations, such as adding nutrient solution or stopping culture.

4) The cell growth density can be intuitively fed back, so that the growth conditions of different cells can be summarized in the process of culturing, different culturing plans are formulated for different cells in different categories, and the cell culturing efficiency is greatly improved.

FIG. 5 is a graph showing the data of 14 days of growth of different cells, and it can be seen from the graph that the straightness is high.

The cell expansion culture and detection method is specifically described below in connection with the cell expansion device and the density sensor of the present utility model, and the specific steps include:

s01: the whole cell expansion device is put into a carbon dioxide incubator together with a fixing plate 1, a cell culture bag 4 is fixed to an X-axis swing pallet 5, a tube of the cell culture bag 4 is fixed to a peristaltic pump/pinch valve 6, and a cell density detecting port 41 of the cell culture bag 4 is fixed to a density sensor 3.

S02: when the machine is started, the electric control board 8 is electrified, the Y-axis motor 7 and the X-axis motor 10 return to zero positions, namely the X-axis swinging supporting plate 5 and the Y-axis bracket 2 return to the complete horizontal position, the weighing sensor 9 reads the weight, and the whole machine is ready.

S03: when culturing cells, the coated cells are injected into the cell culture bag 4, and the horizontal stationary culture is completed for a certain time.

S04: after horizontal resting, according to the needs of a user, a peristaltic pump/pinch valve 6 is started to inject culture medium liquid, and according to the swing amplitude and angle set by the user, a Y-axis motor 7 and/or an X-axis motor 10 can be started simultaneously or singly, and 1 or 2 directions of swing can be generated on an X-axis swing supporting plate 5, so that gentle swing mixing of cells and culture medium liquid is realized, and carbon dioxide permeated by a breathable cell culture bag 4 is mixed into the bag.

S05: starting a density sensor 3, recording cell density data in real time, uploading the data to an electric control board 8, and expanding a large number of cells after the cells grow to a certain number of days; the peristaltic pump/pinch valve 6 is started again according to the needs/program settings of the user to inject the same or different culture medium liquid, and the culture medium liquid and the cells are continuously mixed again according to the new density setting/preset swing amplitude and angle (or swing amplitude and angle before maintaining) and frequency, so that the cells are fully mixed with the culture medium liquid and carbon dioxide, and nutrition growth and amplification are absorbed.

S06: after the cultivation is completed, the data of the weighing sensor 9 is read to obtain the accurate cell weight.

Further, as shown in FIG. 6, using the device and method of the present utility model, specific examples of cell expansion culture were performed from day 5 to day 11, taking coated NK cells as an example.

It will be apparent to those skilled in the art that various modifications and variations can be made to the embodiments of the present utility model without departing from the spirit or scope of the utility model. Thus, if such modifications and variations of the present patent fall within the scope of the patent claims and their equivalents, the present patent is intended to include such modifications and variations.

Claims (10)

1. A device for cell expansion and non-contact real-time measurement of cell expansion density for cell expansion and detection of the density of an analyte in a cell culture bag, comprising:

the weighing sensor is arranged on the fixed plate and used for monitoring weight in real time;

the Y-axis swinging mechanism is arranged on the weighing sensor and is used for swinging along the Y-axis direction;

the X-axis swinging mechanism is arranged on the Y-axis swinging mechanism and is used for swinging along the X-axis direction;

the density sensor is arranged on the X-axis swinging mechanism and used for detecting the density of an object to be detected in the cell culture bag.

2. The device according to claim 1, wherein the cell culture bag is provided with a cell density detection port, and the inside of the cell density detection port is communicated with a space in the cell culture bag.

3. The apparatus of claim 2, wherein the density sensor comprises an infrared light source, a beam splitting prism, a reference infrared receiver a, a detection infrared receiver B, and an optical signal processing circuit.

4. A device according to claim 3, wherein the infrared light source emits parallel rays of 800-1100nm infrared light, and the beam splitting prism is 1: the device comprises a 1 beam splitting prism, wherein infrared parallel light rays are divided into reference parallel light rays and detection parallel light rays after passing through the beam splitting prism, the reference parallel light rays are detected by a reference infrared receiver A, and the detection parallel light rays pass through a cell density detection port and are detected by a detection infrared receiver B.

5. The device of claim 4, wherein the reference parallel light signal and the absorbed parallel light signal enter an optical signal processing circuit, and the illumination values of the reference parallel light signal and the absorbed parallel light signal are compared and corrected to calculate an optical absorbance value, and the optical absorbance value characterizes the density of the cell, so that the change of the density value of the measured object is indirectly detected.

6. The device according to claim 1, wherein the Y-axis swinging mechanism comprises a Y-axis bracket, first swinging shafts positioned at two sides of the Y-axis bracket, and a connecting rod, a first driving rod and a Y-axis motor for driving at least one side of the first swinging shafts to rotate, the two sides of the weighing sensor are provided with supporting frames, the supporting frames are provided with first supporting shaft holes for supporting the first swinging shafts, the first swinging shafts are hinged with one ends of the connecting rod, and the other ends of the connecting rod are hinged with the first driving rod of the Y-axis motor.

7. The device according to claim 6, wherein the X-axis swinging mechanism comprises an X-axis swinging support plate, a second swinging shaft positioned at one side or two sides of the X-axis swinging support plate, a second driving rod for driving the X-axis swinging support plate to swing, and an X-axis motor, wherein the second swinging shaft is arranged in a second supporting shaft hole perpendicular to the first swinging shaft on the Y-axis support in a penetrating way, and the second driving rod is hinged at the bottom of the X-axis swinging support plate and is deviated from the axis where the second swinging shaft is positioned.

8. The device of claim 7, wherein the Y-axis support is a rectangular frame, and the X-axis swing support plate is disposed in the middle of the Y-axis support with gaps around the Y-axis support.

9. The apparatus of claim 7 or 8, wherein the Y-axis motor and the X-axis motor are electrically connected to an electronic control board to control the rotational amplitude and frequency of the Y-axis motor and the X-axis motor.

10. The device according to claim 7, wherein a control valve is arranged on the X-axis swing supporting plate and is used for controlling the connection and disconnection of the cell culture bag and the culture medium liquid pipeline, and the control valve is a peristaltic pump or a pinch valve.