CN114107194B - Immune cell expansion culture method and device - Google Patents
Immune cell expansion culture method and device Download PDFInfo
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- CN114107194B CN114107194B CN202111387469.1A CN202111387469A CN114107194B CN 114107194 B CN114107194 B CN 114107194B CN 202111387469 A CN202111387469 A CN 202111387469A CN 114107194 B CN114107194 B CN 114107194B
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- fixed
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- 210000002865 immune cell Anatomy 0.000 title claims abstract description 21
- 230000010261 cell growth Effects 0.000 title claims description 12
- 238000012136 culture method Methods 0.000 title abstract description 9
- 238000004113 cell culture Methods 0.000 claims abstract description 43
- 230000007246 mechanism Effects 0.000 claims description 10
- 230000000149 penetrating effect Effects 0.000 claims description 6
- 238000000034 method Methods 0.000 abstract description 8
- 230000008569 process Effects 0.000 abstract description 6
- 230000003321 amplification Effects 0.000 abstract description 5
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 5
- 238000007789 sealing Methods 0.000 abstract description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000003028 elevating effect Effects 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 238000005299 abrasion Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000010370 cell cloning Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
-
- 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
- C12M23/04—Flat or tray type, drawers
-
- 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
- C12M23/46—Means for fastening
Abstract
The invention relates to the technical field of cell culture accessory devices, in particular to an immune cell amplification culture method and device, wherein when cell culture is carried out, a sealing door of a culture instrument can not move a cell culture plate in the opening and closing process and the process of a culture plate entering and exiting the culture instrument, so that the culture solution of each hole in the culture plate is prevented from splashing outwards; comprising the following steps: the incubator is hollow in the interior and is provided with an opening at one side, a door is rotatably installed at the opening through a hinge, two bases are fixed in the incubator, a telescopic sliding rail is fixed on each base, a moving plate is fixed on the telescopic ends of the two telescopic sliding rails, two strip holes I are formed in the moving plate in a circumferential array by taking the central line of the moving plate as an axis, a sliding block is slidably installed at one position of each strip hole, and the two sliding blocks synchronously and reversely move to clamp and release the cell culture plate; the movable plate is rotatably provided with a central shaft which is coaxial with the central line of the movable plate, and a first gear is fixed on the central shaft.
Description
Technical Field
The invention relates to the technical field of cell culture accessory devices, in particular to an immune cell amplification culture method and device.
Background
Cell culture refers to a method of simulating in vitro an in vivo environment (sterile, suitable temperature, ph, certain nutritional conditions, etc.) to survive, grow, reproduce and maintain major structures and functions. Cell culture is also known as cell cloning technology, a formal biological term being cell culture technology.
When immune cells are subjected to amplification culture, the immune cells are required to be sorted and then placed in a porous cell culture plate, and are cultured in a culture instrument under the conditions of 37 ℃ and 5% carbon dioxide, wherein the sealing door of the culture instrument can shift in the opening and closing process and the process that the culture plate enters and exits the culture instrument, the culture solution in each hole in the culture plate can be splashed outwards, and the immune cells are mostly grown in a suspension mode and are easy to spill along with the culture solution, so that the culture effect is affected.
Disclosure of Invention
In order to solve the technical problems, the invention provides an immune cell expansion culture method and device, when cell culture is carried out, a sealing door of a culture instrument can not move a cell culture plate in the opening and closing process and the process of entering and exiting the culture instrument, so that culture solution in each hole in the culture plate is prevented from splashing outwards.
The invention relates to an immune cell expansion culture device, which comprises: the incubator is hollow in the interior and is provided with an opening at one side, a door is rotatably installed at the opening through a hinge, two bases are fixed in the incubator, a telescopic sliding rail is fixed on each base, a moving plate is fixed on the telescopic ends of the two telescopic sliding rails, two strip holes I are formed in the moving plate in a circumferential array by taking the central line of the moving plate as an axis, a sliding block is slidably installed at one position of each strip hole, and the two sliding blocks synchronously and reversely move to clamp and release the cell culture plate;
a central shaft is rotatably arranged on the moving plate and is coaxial with the central line of the moving plate, a first gear is fixed on the central shaft, two strip holes which are not concentric with the first gear are formed in a circumferential array on the first gear, a first driving shaft is fixed on each sliding block, and the first driving shaft extends into the second strip holes;
a hollow shaft is rotatably arranged on the moving plate, a gear II meshed with the gear I is fixed on the hollow shaft, a through hole I is formed in the hollow shaft, the through hole I is sequentially provided with a spiral section and a vertical section I from top to bottom, a fixed plate is fixed on one side of the inside of the incubator, a through hole II is formed in the fixed plate, and the through hole II is sequentially provided with a vertical section II and an inclined section from top to bottom from outside to inside;
the lifting mechanism is fixed on the moving plate, the lifting block is installed at the output end of the lifting mechanism, one end of the lifting block is fixed with a second driving shaft penetrating through the spiral section, and the other end of the lifting block is fixed with a third driving shaft penetrating through the second vertical section.
Further, elevating system includes the door template with movable plate fixed connection, rotates on the door template and installs the lead screw, elevating block and lead screw threaded connection are fixed with two guide bars on the door template, and two guide bars all pass the elevating block, installs the motor that is used for driving its rotation on the tip position of lead screw.
Further, a bearing is fixed on each first driving shaft, and the outer ring of the bearing is tangent to the inner wall of the second long strip hole.
Further, a proximity switch for controlling the motor is installed inside the incubator.
Further, soft cushions are arranged on one sides of the two sliding blocks, which are close to the center line of the moving plate.
Further, the height of the bottom ends of the vertical sections is lower than that of the bottom ends of the spiral sections.
Further, a support leg is arranged at each of four corners of the bottom of the incubator.
The invention relates to an immune cell amplification culture method, which comprises the following steps:
s1, extracting peripheral blood mononuclear cells from human peripheral blood;
s2, inoculating the cells extracted in the step S1 into a culture bottle containing a culture medium for culture;
s3, introducing the cells obtained after the culture flask in the step S2 into a culture bag;
s4, obtaining immune cells.
Further, the culturing method in the step S2 comprises the following steps: the flask is placed on a moving plate as claimed in claims 1 to 7 and the moving plate is moved into the incubator as claimed in claims 1 to 7 at 37℃with saturated humidity, 5% CO 2 Culturing in incubator for 3-5 days.
Compared with the prior art, the invention has the beneficial effects that: before the cell culture plate enters the incubator, the device can clamp the cell culture plate or other cell culture devices, then move the cell culture plate or other cell culture devices, then close the chamber door, and when the cell culture plate or other cell culture devices are taken out, open the chamber door and move the cell culture plate or other cell culture devices out of the incubator, the two sliders clamp the cell culture plate or other cell culture devices all the time, so that the culture solution is prevented from being spilled.
Drawings
FIG. 1 is a schematic diagram of the structure of the present invention;
FIG. 2 is a top perspective view of the right front side of FIG. 1;
FIG. 3 is a top perspective view of the left front hidden incubator of FIG. 1 with the rear of the cabinet door;
FIG. 4 is an exploded view of FIG. 3;
FIG. 5 is a block diagram of a hollow shaft, a helical section and a vertical section one;
the reference numerals in the drawings: 1. an incubator; 2. a door; 3. a base; 4. a telescopic slide rail; 5. a moving plate; 6. a strip hole I; 7. a slide block; 8. a central shaft; 9. a first gear; 10. a strip hole II; 11. a first driving shaft; 12. a hollow shaft; 13. a second gear; 14. a helical section; 15. a first vertical section; 16. a fixing plate; 17. a second vertical section; 18. an inclined section; 19. a lifting block; 20. a second driving shaft; 21. a third driving shaft; 22. a screw rod; 23. a guide rod; 24. a motor; 25. a bearing; 26. a proximity switch; 27. and a soft cushion.
Detailed Description
The following describes in further detail the embodiments of the present invention with reference to the drawings and examples. The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
In the description of the present invention, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted", "connected" and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
As shown in fig. 1 to 5, the immune cell expansion culture method and device of the present invention comprises: the incubator comprises an incubator 1, wherein the inside of the incubator is hollow, one side of the incubator is provided with an opening, a refrigerator door 2 is rotatably installed at the opening through a hinge, two bases 3 are fixed in the incubator 1, a telescopic sliding rail 4 is fixed on each base 3, a moving plate 5 is fixed on the telescopic ends of the two telescopic sliding rails 4, two strip holes I6 are formed in the moving plate 5 in a circumferential array by taking the central line of the moving plate 5 as an axis, a sliding block 7 is slidably installed at each strip hole I6, and the two sliding blocks 7 synchronously and reversely move to clamp and release a cell culture plate;
a central shaft 8 is rotatably arranged on the moving plate 5, the central shaft 8 is coaxial with the central line of the moving plate 5, a first gear 9 is fixed on the central shaft 8, two strip holes 10 which are not concentric with the first gear 9 are formed in a circumferential array on the first gear 9, a first driving shaft 11 is fixed on each sliding block 7, and the first driving shaft 11 extends into the strip holes 10;
the movable plate 5 is rotatably provided with a hollow shaft 12, a gear II 13 meshed with the gear I9 is fixed on the hollow shaft 12, a through hole I is formed in the hollow shaft 12, the through hole I is sequentially provided with a spiral section 14 and a vertical section I15 from top to bottom, one side of the inside of the incubator 1 is fixedly provided with a fixed plate 16, the fixed plate 16 is provided with a through hole II, and the through hole II is sequentially provided with a vertical section II 17 and an inclined section 18 from top to bottom from outside to inside;
a lifting mechanism is fixed on the moving plate 5, a lifting block 19 is installed at the output end of the lifting mechanism, a second driving shaft 20 penetrating through the spiral section 14 is fixed at one end of the lifting block 19, and a third driving shaft 21 penetrating through the second vertical section 17 is fixed at the other end of the lifting block 19;
in this embodiment, the condition is that the movable plate 5 is located at the outermost position relative to the incubator 1, when in use, the cell culture plate is placed between the two sliders 7, then the lifting mechanism is operated to move the lifting block 19, the second driving shaft 20 and the third driving shaft 21 downwards at the same time, when the second driving shaft 20 passes through the spiral section 14 of the hollow shaft 12, the third driving shaft 21 moves downwards along the vertical section 17 of the fixed plate 16, and the hollow shaft 12 and the second gear 13 rotate, as the first gear 9 is meshed with the second gear 13, the first gear 9 and the central shaft 8 rotate, and as the first rotating gear 9 enables the two sliders 7 to move towards each other to clamp the cell culture plate, then the second driving shaft 20 moves to the first vertical section 15, the second driving shaft 20 which continues to move downwards does not affect the hollow shaft 12, and the third driving shaft 21 moves to the inclined section 18 of the fixed plate 16, as the fixed plate 16 is fixed in position, the third driving shaft 21 which moves downwards enables the movable plate 5, the lifting mechanism and the second gear 13 to move into the inside the incubator door 1, and then the cell culture plate 2 is closed, and then the cell culture plate is completely moved into the incubator door 2, or the cell culture plate is closed after the cell culture plate is completely inside the incubator door 1, and the cell culture plate is closed, or the cell culture device is closed after the cell culture plate is completely inside the incubator door 2 is closed, and the cell culture plate is opened, and the cell culture device is closed 2 is opened, or the cell culture device is closed when the cell culture device is opened, the two sliders 7 always clamp the cell culture plate or other cell culture devices to prevent the culture solution from spilling.
As a preferable scheme of the embodiment, the lifting mechanism comprises a door-shaped plate fixedly connected with the movable plate 5, a screw rod 22 is rotatably arranged on the door-shaped plate, the lifting block 19 is in threaded connection with the screw rod 22, two guide rods 23 are fixed on the door-shaped plate, the two guide rods 23 penetrate through the lifting block 19, and a motor 24 for driving the screw rod 22 to rotate is arranged at the end part of the screw rod 22;
in this embodiment, after the motor 24 is energized, the screw 22 is driven to rotate, and because the screw 22 is in threaded connection with the lifting block 19 and the lifting block 19 cannot rotate under the action of the two guide rods 23, rotating the screw 22 can change the height of the lifting block 19, and the motor 24 is stopped by adopting a driving mode of the screw 22, the self-locking function is achieved.
As a preferable scheme of the embodiment, a bearing 25 is fixed on each first driving shaft 11, and the outer ring of the bearing 25 is tangent to the inner wall of the second elongated hole 10;
in this embodiment, by installing the bearing 25 on the driving shaft for avoiding the driving shaft from contacting the inner wall of the second elongated hole 10, the abrasion to the driving shaft can be prevented to ensure the synchronism of the movement of the two sliders 7.
As a preferable scheme of the above embodiment, the incubator 1 is internally provided with a proximity switch 26 for controlling the motor 24;
in the present embodiment, when the door type plate moves to the inside of the incubator 1 to trigger the proximity switch 26, the proximity switch 26 controls the motor 24 to be turned off, so that the motor 24 does not need to be manually controlled, and the moving plate 5 is prevented from colliding with the incubator 1.
As a preferable scheme of the above embodiment, the cushion 27 is mounted on one side of the two sliding blocks 7 close to the center line of the moving plate 5;
in this embodiment, the soft pad 27 on the slider 7 is in direct contact with the cell culture device, the soft pad 27 deforms during clamping, applicability is improved, and the soft pad 27 does not damage the cell culture device.
As a preferable scheme of the embodiment, the bottom end height of the second vertical section 17 is lower than the bottom end height of the spiral section 14;
in this embodiment, the movable plate 5 is moved into the incubator 1 after the cell culture apparatus is clamped.
As a preferable scheme of the embodiment, four corners of the bottom of the incubator 1 are provided with one supporting leg;
in this embodiment, the incubator 1 is supported by the provided legs.
The invention relates to an immune cell amplification culture method, which comprises the following steps:
s1, extracting peripheral blood mononuclear cells from human peripheral blood;
s2, inoculating the cells extracted in the step S1 into a culture bottle containing a culture medium for culture;
s3, introducing the cells obtained after the culture flask in the step S2 into a culture bag;
s4, obtaining immune cells.
As a preferable mode of the above embodiment, the culture method of step S2 is as follows: the flask was placed on the moving plate 5 of claims 1 to 7, and the moving plate 5 was moved to the inside of the incubator 1 of claims 1 to 7, and incubated in the incubator 1 of 5% CO2 at 37℃and saturated humidity for 3 to 5 days.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that it will be apparent to those skilled in the art that modifications and variations can be made without departing from the technical principles of the present invention, and these modifications and variations should also be regarded as the scope of the invention.
Claims (7)
1. An immune cell expansion culture device, comprising: the incubator comprises an incubator body (1), wherein the inside of the incubator body is hollow, one side of the incubator body is provided with an opening, a refrigerator door (2) is rotatably installed at the opening through a hinge, two bases (3) are fixed inside the incubator body (1), a telescopic sliding rail (4) is fixed on each base (3), a moving plate (5) is fixed on the telescopic ends of the two telescopic sliding rails (4), two strip holes I (6) are formed in the moving plate (5) in a circumferential array by taking the central line of the moving plate (5) as an axis, a sliding block (7) is slidably installed at each strip hole I (6), and the two sliding blocks (7) synchronously and reversely move to clamp and release a cell culture plate;
a central shaft (8) is rotatably arranged on the movable plate (5), the central shaft (8) is coaxial with the central line of the movable plate (5), a first gear (9) is fixed on the central shaft (8), two long-strip holes (10) which are not concentric with the first gear (9) are formed in the first gear (9) in a circumferential array, a first driving shaft (11) is fixed on each sliding block (7), and the first driving shaft (11) extends into the long-strip holes (10);
a hollow shaft (12) is rotatably arranged on the moving plate (5), a gear II (13) meshed with the gear I (9) is fixed on the hollow shaft (12), a through hole I is formed in the hollow shaft (12), a spiral section (14) and a vertical section I (15) are sequentially arranged on one side of the inside of the incubator (1) from top to bottom, a fixed plate (16) is fixed on one side of the inside of the incubator (1), a through hole II is formed in the fixed plate (16), and a vertical section II (17) and an inclined section (18) are sequentially arranged on the through hole II from top to bottom;
the lifting mechanism is fixed on the moving plate (5), the lifting block (19) is installed at the output end of the lifting mechanism, a driving shaft II (20) penetrating through the spiral section (14) is fixed at one end of the lifting block (19), and a driving shaft III (21) penetrating through the vertical section II (17) is fixed at the other end of the lifting block.
2. An immune cell expansion culture device according to claim 1, characterized in that the lifting mechanism comprises a door-shaped plate fixedly connected with the movable plate (5), a screw rod (22) is rotatably arranged on the door-shaped plate, the lifting block (19) is in threaded connection with the screw rod (22), two guide rods (23) are fixed on the door-shaped plate, the two guide rods (23) penetrate through the lifting block (19), and a motor (24) for driving the screw rod (22) to rotate is arranged at the end part of the screw rod.
3. An immune cell expansion culture device according to claim 2, wherein a bearing (25) is fixed on each first driving shaft (11), and the outer ring of the bearing (25) is tangent to the inner wall of the second elongated hole (10).
4. An immune cell expansion culture device according to claim 3, characterized in that the incubator (1) is internally provided with a proximity switch (26) for controlling the motor (24).
5. An immune cell expansion culture device according to claim 4, characterized in that the two sliders (7) are each provided with a cushion (27) on the side close to the center line of the moving plate (5).
6. An immune cell expansion culture device according to claim 5, wherein the bottom end height of the second vertical section (17) is lower than the bottom end height of the spiral section (14).
7. An immune cell expansion culture device according to claim 6, wherein a support leg is arranged at each of four corners of the bottom of the incubator (1).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111387469.1A CN114107194B (en) | 2021-11-22 | 2021-11-22 | Immune cell expansion culture method and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111387469.1A CN114107194B (en) | 2021-11-22 | 2021-11-22 | Immune cell expansion culture method and device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114107194A CN114107194A (en) | 2022-03-01 |
| CN114107194B true CN114107194B (en) | 2024-01-30 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202111387469.1A Active CN114107194B (en) | 2021-11-22 | 2021-11-22 | Immune cell expansion culture method and device |
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| CN (1) | CN114107194B (en) |
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| CN212199289U (en) * | 2019-11-13 | 2020-12-22 | 北京易科拜德科技有限公司 | Cell culture device |
| CN212925056U (en) * | 2020-07-17 | 2021-04-09 | 威海见生生物技术有限公司 | Immune cell culture device |
| CN214004663U (en) * | 2020-11-12 | 2021-08-20 | 南昌龙来福生物科技有限公司 | Biological cell constant temperature incubator |
| CN214060537U (en) * | 2020-12-04 | 2021-08-27 | 冬青南京生物科技有限公司 | Immune cell amplification culture instrument |
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2021
- 2021-11-22 CN CN202111387469.1A patent/CN114107194B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2014171389A (en) * | 2013-03-05 | 2014-09-22 | Shimadzu Corp | Cell culture device |
| CN109207424A (en) * | 2018-10-24 | 2019-01-15 | 河南省肿瘤医院 | A kind of cultural method of immunocyte |
| CN210736784U (en) * | 2019-07-30 | 2020-06-12 | 南京平港生物技术有限公司 | Cell culture box |
| CN212199289U (en) * | 2019-11-13 | 2020-12-22 | 北京易科拜德科技有限公司 | Cell culture device |
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| CN214004663U (en) * | 2020-11-12 | 2021-08-20 | 南昌龙来福生物科技有限公司 | Biological cell constant temperature incubator |
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| CN114107194A (en) | 2022-03-01 |
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