CN111197006B - CAR-T cell expander - Google Patents
CAR-T cell expander Download PDFInfo
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- CN111197006B CN111197006B CN202010047797.6A CN202010047797A CN111197006B CN 111197006 B CN111197006 B CN 111197006B CN 202010047797 A CN202010047797 A CN 202010047797A CN 111197006 B CN111197006 B CN 111197006B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/16—Vibrating; Shaking; Tilting
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- Bioinformatics & Cheminformatics (AREA)
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Abstract
The invention discloses a CAR-T cell expander, comprising: a main body, a culturing bag, a cell refrigerator for refrigerating a culturing reagent, and a pipe transfer system capable of transferring a liquid in the cell refrigerator to the culturing bag; the pipeline conveying system comprises a peristaltic pump for pumping the culture reagent from the cell refrigerator into the culture bag; the CAR-T cell expander further comprises: a pipeline heating module for heating the culture reagent conveyed by the pipeline conveying system to room temperature; the cell refrigerator, the peristaltic pump and the pipeline heating module are all installed to the main machine body. The invention has the advantages that the CAR-T cells can be amplified and cultured, the cell refrigerator is used for refrigerating the culture reagent, and the temperature of the culture reagent can be raised through the pipeline heating module if the culture reagent is needed.
Description
Technical Field
The invention relates to a CAR-T cell expander.
Background
T cells, also called T lymphocytes, are one of the human leukocytes, derived from peripheral blood, and exert an immune function in humans. The effect is equivalent to that of a fighter in a human body, and the fighter can resist and eliminate enemy such as infection, tumor, foreign matters and the like. In the laboratory, technicians activate T cells by genetic engineering techniques and equip with a localization navigation device CAR (tumor chimeric antigen receptor) to modify the general "fighter" T cells into "super fighter", i.e., CAR-T cells.
The traditional CAR-T cell expansion adopts an artificial culture mode, and is complex in steps and large in workload.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides the CAR-T cell expander which can expand and culture CAR-T cells, and the cell refrigerator stores culture reagents at low temperature, and if the culture reagents are needed, the culture reagents can be heated by a pipeline heating module.
In order to achieve the above object, the present invention adopts the following technical scheme:
a CAR-T cell expander comprising: a main body, a culturing bag, a cell refrigerator for refrigerating a culturing reagent, and a pipe transfer system capable of transferring a liquid in the cell refrigerator to the culturing bag; the pipeline conveying system comprises a peristaltic pump for pumping the culture reagent from the cell refrigerator into the culture bag; the CAR-T cell expander further comprises: a pipeline heating module for heating the culture reagent conveyed by the pipeline conveying system to room temperature; the cell refrigerator, the peristaltic pump and the pipeline heating module are all installed to the main machine body.
Further, the main body is formed with a culture cavity; the CAR-T cell expander further comprises a shaking device for shaking the culture bag; the shaking device and the culture bag are arranged in the culture cavity.
Further, the shaking device includes: the culture device comprises a base, a tray for bearing culture bags, an upper transmission assembly, a middle transmission assembly and a lower transmission assembly; the lower transmission assembly includes: the first movable seat, the first motor and the eccentric block; the eccentric block is fixed to a motor shaft of the first motor; the eccentric block is connected to the first movable seat; the first motor drives the first movable seat to revolve around a motor shaft of the first motor through the eccentric block; the middle transmission assembly comprises: the second movable seat, the second motor, the second cam and the return spring are used for pushing the second movable seat in the horizontal direction; the first movable seat is provided with a sliding rail for guiding the second movable seat to slide and a positioning seat for positioning the return spring; the second motor drives the second cam to rotate and pushes the second movable seat to slide along the sliding rail through the second cam; two ends of the reset spring are connected with the positioning seat and the second movable seat; the reset spring and the second cam are positioned at two ends of the second movable seat; the upper transmission assembly includes: the third motor is used for pushing the tray along the height direction to enable the tray to move in a fluctuating manner along the height direction, and the plurality of connecting springs are connected with the tray and the second movable seat; one end of the connecting spring is fixed to the bottom of the tray; the other end of the connecting spring is fixed to the top of the second movable seat; the third motor drives the third cam to rotate; the third cam is in contact with the bottom of the tray.
Further, the lower transmission assembly further comprises: a plurality of connecting ropes; the connecting rope is connected with the base and the first movable seat; a plurality of connecting ropes encircle the motor shaft of the first motor.
Further, the eccentric block is provided with an eccentric cylinder; the axis of the eccentric cylinder is offset from the rotational axis of the motor shaft of the first motor.
Further, the first movable seat is formed with a bearing seat; the lower transmission assembly further comprises a bearing; the bearing is mounted to the bearing seat; the bearing is sleeved on the periphery of the eccentric cylinder.
Further, the upper transmission assembly further comprises: a driving shaft and a driven shaft; the number of the third cams is even; the plurality of third cams are equally divided into two groups which are respectively arranged on the driving shaft and the driven shaft; the third motor drives the driving shaft to rotate and drives the driven shaft to rotate through belt pulley transmission.
Further, the third cams on the driving shaft and the driven shaft are circumferentially spaced 180 degrees apart when rotated.
Further, when the eccentric convex structure of the third cam on the driving shaft jacks up the tray, the eccentric convex structure of the third cam on the driven shaft is not contacted with the tray; when the eccentric convex structure of the third cam on the driven shaft jacks up the tray, the eccentric convex structure of the third cam on the driving shaft is not contacted with the tray.
Further, the number of the connecting springs is 4; the 4 connecting springs are arranged in a rectangular shape; the third cam is located within the rectangle.
The invention has the advantages that the CAR-T cells can be amplified and cultured, the culture reagent can be put into a cell refrigerator for refrigeration during storage, and if the culture reagent is needed to be used, the culture reagent can be heated by a pipeline heating module, so that the temperature is convenient to raise.
The mode that adopts the device of swaying to cultivate the bag to shake sways, compares traditional stirring mode shearing force little, and is little to the cell damage. The uniform distribution of the nutrient components can be ensured by shaking, and the shearing force can be reduced, so that the damage to cells is reduced.
The shaking device shakes the culture bag from multiple dimensions, so that the distribution of nutrient components is more uniform compared with a single-shaking mode.
Drawings
FIG. 1 is a schematic diagram of a CAR-T cell expander according to the present invention;
FIG. 2 is a schematic diagram of a shaking device of the CAR-T cell expander of FIG. 1;
FIG. 3 is a cross-sectional view of the rocking device of FIG. 2;
fig. 4 is a schematic view showing the installation of the driving shaft and the third cam of the swing apparatus of fig. 3.
The CAR-T cell expander 100, the main body 10, the culture chamber 11, the cell refrigerator 20, the peristaltic pump 30, the pipeline heating module 40, the shaking device 50, the base 51, the tray 52, the upper transmission assembly 53, the third cam 531, the connecting spring 532, the driving shaft 533, the driven shaft 534, the middle transmission assembly 54, the second movable seat 541, the second cam 542, the return spring 543, the lower transmission assembly 55, the first movable seat 551, the positioning seat 551a, the bearing seat 551b, the first motor 552, the eccentric block 553, the eccentric cylinder 553a, the connecting rope 554, and the bearing 555.
Detailed Description
The invention is described in detail below with reference to the drawings and the specific embodiments.
As shown in fig. 1-4, a CAR-T cell expander 100 comprises: a main body 10, a culture bag, a cell refrigerator 20 and a pipeline transportation system. The cell freezer 20 is used to chill culture reagents. The tubing transport system is capable of transporting the liquid within the cell freezer 20 to the culture bag. The tubing delivery system includes peristaltic pump 30. Peristaltic pump 30 is used to draw culture reagents from within cell freezer 20 and pump the culture bags. Specifically, peristaltic pump 30 provides a pumping function for the liquid and, when activated, allows the culture reagent to pass from cell freezer 20 to culture chamber 11. The tubing delivery system further includes a hose. The hose is used for conveying the culture reagent. The CAR-T cell expander 100 further comprises: a pipeline heating module 40. The pipeline heating module 40 is used for heating the culture reagent conveyed by the pipeline conveying system to room temperature. The cell refrigerator 20, peristaltic pump 30 and line heating module 40 are all mounted to the main body 10.
The CAR-T cell expander 100 adopts the cell refrigerator 20 to store the culture reagent, and the culture reagent is put into the cell refrigerator 20 for refrigeration, and can be heated by a pipeline heating module if the culture reagent is needed, so that the temperature is convenient to rise.
The cell refrigerator is characterized in that the temperature range is 2-8 ℃ and the control precision is +/-1 ℃ relative to a refrigerator, and the cell refrigerator is used for refrigerating and culturing the reagent.
As a specific implementation mode, the pipeline heating module consists of a heating plate, a heating mounting plate and a temperature sensor, wherein the heating plate is positioned between the reagent culture chamber and the cell culture chamber, and the pipeline heating module has the function of heating liquid penetrating out of the pipeline from 2-8 ℃ to room temperature. The heating pin is mounted to the heating mounting plate. The temperature sensor detects the temperature. The heating plate is electrified to heat the reagent.
As a specific embodiment, the CAR-T cell expander 100 further comprises: and a control panel. The control panel is an electronic screen, is controlled by touch, contains equipment control software and can set parameters required by the amplification culture process.
As a specific embodiment, the main body 10 is formed with a culture chamber 11. The CAR-T cell expander 100 further comprises a shaking device 50. The shaking device 50 is used to shake the culture bag. The shaking device 50 and the culture bag are provided in the culture chamber 11.
Specifically, the culture chamber 11 is a cell culture position, and is externally connected with a carbon dioxide tank, so that the temperature, the carbon dioxide concentration, the humidity and the like can be controlled. A shaking device 50 is provided in the culture bag to periodically shake the culture bag to prevent cell deposition.
As a specific embodiment, the shaking device 50 includes: base 51, tray 52, upper drive assembly 53, middle drive assembly 54, and lower drive assembly 55. The tray 52 is used to carry the culture bag.
Specifically, the lower transmission assembly 55 includes: the first movable seat 551, the first motor 552 and the eccentric block 553. The eccentric mass 553 is fixed to the motor shaft of the first motor 552. The eccentric mass 553 is connected to the first movable seat 551. The first motor 552 drives the first movable seat 551 to revolve around the motor shaft of the first motor 552 through the eccentric block 553. The lower transmission assembly 55 further includes: a plurality of connecting strings 554. The connection rope 554 connects the base 51 and the first movable seat 551. A plurality of connecting ropes 554 wrap around a motor shaft of the first motor 552.
The middle drive assembly 54 includes: a second movable seat 541, a second motor, a second cam 542, and a return spring 543. The second cam 542 serves to push the second movable seat 541 in the horizontal direction. The first movable seat 551 is provided with a sliding rail and a positioning seat 551a. The slide rail guides the second movable seat 541 to slide. The positioning seat 551a is used for positioning the return spring 543. The sliding rail is disposed at the top of the first movable seat 551. The second motor drives the second cam 542 to rotate, and the second movable seat 541 is pushed to slide along the sliding rail by the second cam 542. Both ends of the return spring 543 are connected to the positioning seat 551a and the second movable seat 541. The return spring 543 and the second cam 542 are located at both ends of the second movable seat 541.
The upper transmission assembly 53 includes: a third motor, a third cam 531, and a plurality of connection springs 532. The third cams 531 serve to push the tray 52 in the height direction to move the tray 52 in the height direction in a fluctuating manner. The connection spring 532 connects the tray 52 and the second movable seat 541. One end of the connection spring 532 is fixed to the bottom of the tray 52. The other end of the connection spring 532 is fixed to the top of the second movable seat 541. The third motor drives the third cam 531 to rotate. The third cam 531 contacts the bottom of the tray 52. The upper transmission assembly 53 further includes: a drive shaft 533 and a follower shaft 534. The number of the third cams 531 is even. The plurality of third cams 531 are equally divided into two groups mounted to the driving shaft 533 and the driven shaft 534, respectively. The third motor drives the driving shaft 533 to rotate and drives the driven shaft 534 to rotate through the pulley transmission.
Cells are fragile, and the shearing force is large in a traditional stirring mode, so that the damage to the cells is large. To solve the damage caused by the conventional stirring method, the CAR-T cell expander 100 uses a shaking device 50 to shake the culture bag, thereby reducing the number of cells. Specifically, the shaking device 50 is divided into an upper transmission assembly 53, a middle transmission assembly 54, and a lower transmission assembly 55.
In the lower transmission assembly 55, the first motor 552 drives the eccentric block 553, and then the eccentric block 553 drives the first movable seat 551 to revolve around the motor shaft of the first motor 552, and the base 51 and the first movable seat 551 are connected through the connection rope 554, so that the first movable seat 551 can be prevented from rotating while revolving. The flexible pipe winding caused by the rotation of the culture bag can be avoided. A first motor 552 is mounted to the base 51. The first motor 552 drives the first movable seat 551 to move through the eccentric block.
The second motor is mounted to the first movable seat 551. The second cam 542 is rotatably coupled to the first movable seat. In the middle transmission assembly 54, the second motor drives the second cam 542 to rotate first, then the second cam 542 rotates to push one side of the second movable seat 541 in the horizontal direction, and the other side of the second movable seat 541 pushes the second movable seat 541 to contact with the second cam 542 due to the elastic force of the return spring 543, so that the second cam 542 can push the second movable seat 541 to slide left and right along the sliding rail direction when the second motor drives the second cam 542.
The third motor is mounted to the second movable seat 541. The third cam 531 is rotatably connected to the second movable seat 541. In the upper transmission assembly 53, first, the third motor drives the third cam 531 to rotate, and when the third cam 531 on the driving shaft 533 rotates clockwise, the tray 52 moves up and down with the rotation of the third cam 531 on the driving shaft 533 and the third cam 531 on the driven shaft 534 because the third cam 531 on the driving shaft 533 and the third cam 531 on the driven shaft 534 are oriented differently.
Therefore, by using the shaking device 50 to shake the culture bag, the nutrient components can be uniformly distributed, and the shearing force can be reduced, thereby reducing damage to cells. Under the combined action of the upper transmission component 53, the middle transmission component 54 and the lower transmission component 55, the culture bag is swayed in a multi-dimensional scale.
As a specific embodiment, the eccentric mass 553 is provided with an eccentric cylinder 553a. The axis of the eccentric cylinder 553a is offset from the rotational axis of the motor shaft of the first motor 552.
As a specific embodiment, the first movable seat 551 is formed with a bearing seat 551b. The lower drive assembly 55 also includes a bearing 555. Bearing 555 is mounted to bearing mount 551b. The bearing 555 is fitted around the outer circumference of the eccentric cylinder 553a.
As a specific embodiment, the third cams 531 on the driving shaft 533 and the driven shaft 534 are circumferentially spaced apart by 180 degrees when rotated.
As a specific embodiment, when the eccentric protrusion structure of the third cam 531 on the driving shaft 533 lifts up the tray 52, the eccentric protrusion structure of the third cam 531 on the driven shaft 534 is not in contact with the tray 52. When the eccentric protrusion structure of the third cam 531 on the follower shaft 534 lifts up the tray 52, the eccentric protrusion structure of the third cam 531 on the drive shaft 533 does not contact the tray 52.
As a specific embodiment, the number of connection springs 532 is 4. The 4 connection springs 532 are arranged in a rectangular shape. The third cam 531 is located within the rectangle.
The CAR-T cell expander 100 is used as follows: transferring a culture medium to a reagent storage bag through a sample inlet and outlet pipeline under a sterile environment, transferring a cell suspension to a cell culture bag, then arranging the reagent storage bag in a cell refrigerator, and arranging the cell culture bag on a shaking device in a culture cavity; and then connecting the cell culture bag and the reagent storage bag through pipelines, and respectively installing the connected pipelines on a peristaltic pump and a pipeline heating module.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be appreciated by persons skilled in the art that the above embodiments are not intended to limit the invention in any way, and that all technical solutions obtained by means of equivalent substitutions or equivalent transformations fall within the scope of the invention.
Claims (1)
1. A CAR-T cell expander, comprising: a main body, a culture bag, a cell refrigerator for refrigerating a culture reagent, and a pipe transfer system capable of transferring a liquid in the cell refrigerator to the culture bag; the pipeline conveying system comprises a peristaltic pump for pumping culture reagent from the cell refrigerator into the culture bag; the CAR-T cell expander further comprises: a pipeline heating module for heating the culture reagent conveyed by the pipeline conveying system to room temperature; the cell refrigerator, the peristaltic pump and the pipeline heating module are all mounted to the main machine body;
the main body is formed with a culture cavity; the CAR-T cell expander further comprises a shaking device for shaking the culture bag; the shaking device and the culture bag are arranged in the culture cavity;
the shaking device includes: the tray is used for bearing the culture bags, and comprises a base, an upper transmission assembly, a middle transmission assembly and a lower transmission assembly; the lower transmission assembly includes: the first movable seat, the first motor and the eccentric block; the eccentric block is fixed to a motor shaft of the first motor; the eccentric block is connected to the first movable seat; the first motor drives the first movable seat to revolve around a motor shaft of the first motor through the eccentric block;
the middle transmission assembly comprises: the second movable seat, the second motor, the second cam and the return spring are used for pushing the second movable seat in the horizontal direction; the first movable seat is provided with a sliding rail for guiding the second movable seat to slide and a positioning seat for positioning the reset spring; the second motor drives the second cam to rotate, and the second movable seat is pushed by the second cam to slide along the sliding rail; the two ends of the reset spring are connected with the positioning seat and the second movable seat; the return spring and the second cam are positioned at two ends of the second movable seat;
the upper transmission assembly includes: the third motor is used for pushing the tray in the height direction to enable the tray to move in a fluctuating manner in the height direction, and the plurality of connecting springs are connected with the tray and the second movable seat; one end of the connecting spring is fixed to the bottom of the tray; the other end of the connecting spring is fixed to the top of the second movable seat; the third motor drives the third cam to rotate; the third cam is in contact with the bottom of the tray;
the lower transmission assembly further includes: a plurality of connecting ropes; the connecting rope is connected with the base and the first movable seat; a plurality of the connecting ropes encircle a motor shaft of the first motor;
the eccentric block is provided with an eccentric cylinder; the axis of the eccentric cylinder is deviated from the rotation axis of the motor shaft of the first motor;
the first movable seat is provided with a bearing seat; the lower transmission assembly further comprises a bearing; the bearing is mounted to the bearing housing; the bearing is sleeved on the periphery of the eccentric cylinder;
the upper transmission assembly further includes: a driving shaft and a driven shaft; the number of the third cams is even; the plurality of third cams are equally divided into two groups which are respectively mounted to the driving shaft and the driven shaft; the third motor drives the driving shaft to rotate and drives the driven shaft to rotate through belt pulley transmission;
the third cams on the driving shaft and the driven shaft are circumferentially 180 degrees apart when rotating;
when the eccentric protrusion structure of the third cam on the driving shaft jacks up the tray, the eccentric protrusion structure of the third cam on the driven shaft is not in contact with the tray; when the eccentric protrusion structure of the third cam on the driven shaft jacks up the tray, the eccentric protrusion structure of the third cam on the driving shaft is not in contact with the tray;
the number of the connecting springs is 4; the 4 connecting springs are arranged in a rectangular shape; the third cam is located within the rectangle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010047797.6A CN111197006B (en) | 2020-01-16 | 2020-01-16 | CAR-T cell expander |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010047797.6A CN111197006B (en) | 2020-01-16 | 2020-01-16 | CAR-T cell expander |
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| CN111197006A CN111197006A (en) | 2020-05-26 |
| CN111197006B true CN111197006B (en) | 2023-09-12 |
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| CN110628619A (en) * | 2019-10-19 | 2019-12-31 | 陈龙刚 | Cell engineering multidimension degree cell oscillation device |
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| JP2004357523A (en) * | 2003-06-02 | 2004-12-24 | Olympus Corp | Cell-culturing and detecting apparatus |
| CN106566772A (en) * | 2015-10-13 | 2017-04-19 | 刘伦光 | Cell incubator |
| CN105925481A (en) * | 2016-05-16 | 2016-09-07 | 上海吉凯基因科技有限公司 | Full-automatic cell continuous culture system |
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| CN111197006A (en) | 2020-05-26 |
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