CN111378559A - Centrifugal transfection and combined culture integrated consumable suitable for automatic production - Google Patents
Centrifugal transfection and combined culture integrated consumable suitable for automatic production Download PDFInfo
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- CN111378559A CN111378559A CN202010198625.9A CN202010198625A CN111378559A CN 111378559 A CN111378559 A CN 111378559A CN 202010198625 A CN202010198625 A CN 202010198625A CN 111378559 A CN111378559 A CN 111378559A
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- 238000001890 transfection Methods 0.000 title claims abstract description 34
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- 238000007789 sealing Methods 0.000 claims abstract description 49
- 238000003501 co-culture Methods 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 5
- 239000003381 stabilizer Substances 0.000 claims description 5
- 230000010354 integration Effects 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 abstract description 8
- 238000000034 method Methods 0.000 description 11
- 238000005119 centrifugation Methods 0.000 description 10
- 239000007788 liquid Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
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- 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
- C12M23/24—Gas permeable parts
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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
- C12M41/48—Automatic or computerized control
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Abstract
The invention belongs to the technical field of centrifugal transfection and combined culture, and relates to a centrifugal transfection and combined culture integrated consumable suitable for automatic production, which comprises an outer shell, an upper sealing plate and a lower sealing plate, wherein the upper sealing plate is fixedly connected to the upper end surface of the outer shell through a convex shoulder, and the two sides of the upper sealing plate are fixedly provided with the outer sealing plates; the upper sealing plate is provided with a container opening, and a container cover is connected to the container opening in a threaded manner; the lower end of the outer sealing plate is provided with a lower sealing plate, the lower end of the lower sealing plate is uniformly provided with a plurality of rib plates, and the lower end of the outer shell is fixedly provided with a plurality of support legs; the invention solves the problem that the traditional device is easy to pollute after cells are put into a culture medium after centrifugal transfection.
Description
Technical Field
The invention belongs to the technical field of centrifugal transfection and combined culture, and relates to an integrated consumable for centrifugal transfection and combined culture, which is suitable for automatic production.
Background
The biological medicines are in a rapid development stage, and novel cellular immune medicines such as CAR-T, TCR-T and the like are about to be put into large-scale use. Centrifugal transfection is a key process step in the production process of cell drugs. When the cells are transfected to produce the gene-modified cells, a standing or centrifugation method is generally employed. In order to increase the transfection efficiency, centrifugation is preferred for transfection. The traditional 96-well plate transfection mode is small in transfection amount, not suitable for large-scale production and inconvenient for automatic centrifugal transfection operation, and a culture container needs to be replaced immediately after transfection in the traditional manual process, so that the transfection efficiency is reduced. A special consumable is needed, and the method is suitable for batch operation of automatic equipment. Meanwhile, after centrifugal transfection, stationary-phase culture can be carried out in a container for 24-48 hours.
Based on the technical scheme, the invention provides an integrated consumable for centrifugal transfection and combined culture, which is suitable for automatic production and aims to solve the technical problem.
Disclosure of Invention
The invention aims to provide an integrated consumable for centrifugal transfection and combined culture, which is suitable for automatic production and solves the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme:
the utility model provides a centrifugal transfection and co-culture integration consumptive material that is fit for automated production, includes the shell body, goes up the shrouding and down the shrouding, its characterized in that: the upper sealing plate is fixedly connected to the upper end face of the outer shell through a convex shoulder, on one hand, the convex shoulder is convenient to take, on the other hand, the damage rate of the device in the transportation process can be effectively reduced due to the arc design of the convex shoulder, and the outer sealing plates are fixedly arranged on two sides of the upper sealing plate; the upper sealing plate is provided with a container opening, and a container cover is connected to the container opening in a threaded manner; the threaded connection can be better sealed, and the threaded connection can enable the device to be more stable in the centrifugal process; the outer shrouding lower extreme is provided with down the shrouding, the shrouding lower extreme evenly is provided with a plurality of gussets down, cavity and the stabilizer blade of lower extreme between the gusset, and the manipulator firmly snatchs to the device is holistic when making things convenient for the centrifugation, the fixed a plurality of stabilizer blades that are provided with of shell body lower extreme.
In a further aspect of the present invention, the outer sealing plate is provided with a window, and the window on the outer sealing plate is provided to facilitate observation of the culture medium in a sealed state.
As a further scheme of the invention, the windows are distributed in a vertically symmetrical mode, the number of the windows is more than one, and the distribution of a plurality of windows can improve the observation efficiency, increase the number of observed samples and further reduce the observation error.
As a further scheme of the invention, the window is internally and fixedly provided with a waterproof breathable film, and the waterproof breathable film is arranged, so that cell respiration in the culture cavity is facilitated on one hand, and the container cover can be completely sealed conveniently on the other hand, thereby avoiding the pollution to the culture medium caused by the need of ventilating the culture cavity after centrifugation.
As a further scheme of the invention, the anti-overflow flange is fixedly arranged on the surface of the container mouth, so that the sealing efficiency of the container mouth in the centrifugal process is improved.
As a further scheme of the invention, the container cover is a self-sealing container cover, so that the sealing efficiency is improved.
As a further scheme of the present invention, the upper sealing plate and the lower sealing plate are both concave sealing plates. The concave design of the upper sealing plate is beneficial to the culture medium solution which is stained on the inner wall of the upper sealing plate to concentrate along the inclined direction and finally fall into the culture cavity after centrifugation. The concave design of the lower sealing plate helps the pipette to completely draw the culture medium away at the concave point when the medium needs to be pipetted or removed.
As a further aspect of the invention, the pits are a middle portion of the front view. The pipette can conveniently find the lowest liquid level of the culture cavity.
As a further scheme of the invention, the support legs and the rib plates are distributed intermittently in the front view direction. Facilitating better gripping means of the robotic arm.
Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
1. this device overall structure is simple, combines as an organic whole with the cultivation consumptive material through reasonable structural design, realizes the combination of stable cultivation after centrifugal transfection and the transfection, effectively improves the technology integrated level, simplifies the technology complexity.
2. The waterproof breathable film greatly liberates the sealing reliability of the original container cover, improves the transfection efficiency in the transfection process and improves the survival rate of target cells after transfection.
3. Through the arrangement of the positioning cavity, the positioning cavity is more easily combined with an automatic structure, and the robot automatic system is convenient to automatically operate in a large scale; in addition, the centrifugation and the culture are integrated, so that the cost is greatly saved, the resource utilization rate is improved, and the method is more environment-friendly and reliable.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic diagram of the general structure of the present invention;
FIG. 2 is a schematic cross-sectional view of the present invention;
FIG. 3 is a schematic view showing a spatial structure of a container mouth after the container cover of the present invention is cut away;
fig. 4 is a schematic view of the present invention in a substantially planar three-dimensional view.
FIGS. 1-4;
1-container cover, 2-shoulder, 3-container mouth, 4-outer sealing plate, 5-window, 6-rib plate, 7-support leg, 8-positioning cavity, 9-outer shell, 10-waterproof breathable film, 11-culture cavity, 12-upper sealing plate, 13-lower sealing plate and 14-anti-overflow flange.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to fig. 1 to 4 in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-4, a centrifugal transfection and co-culture integrated consumable suitable for automated production includes an outer housing 9, an upper sealing plate 12 and a lower sealing plate 13, wherein: the upper sealing plate 12 is fixedly connected to the upper end face of the outer shell 9 through the convex shoulder 2, on one hand, the convex shoulder 2 is used for facilitating taking, on the other hand, the damage rate of the device in the transportation process can be effectively reduced due to the arc design of the convex shoulder 2, and the outer sealing plates 4 are fixedly arranged on two sides of the upper sealing plate 12; the upper sealing plate 12 is provided with a container opening 3, and the container opening 3 is in threaded connection with a container cover 1; the threaded connection can be better sealed, and the threaded connection can enable the device to be more stable in the centrifugal process; the lower extreme of outer shrouding 4 is provided with down shrouding 13, shrouding 13 lower extreme evenly is provided with a plurality of baffles down, cavity and the stabilizer blade 7 of lower extreme between the baffle, and cavity between shell body 9 and the gusset 6 is location chamber 8, and manipulator firmly snatchs to the device is holistic when making things convenient for the centrifugation, the fixed a plurality of stabilizer blades 7 that are provided with of shell body 9 lower extreme.
Wherein, the outer shrouding 4 is last to be seted up window 5, and the window 5 setting on the outer shrouding 4 helps under the encapsulated situation, to the observation of culture medium. The window 5 is distributed for longitudinal symmetry, and the number is greater than one, and the distribution of a plurality of windows can improve observation efficiency, increases the sample number of observing to reduce observation error. The window 5 internal fixation is provided with waterproof ventilated membrane 10, and the setting of waterproof ventilated membrane 10 is favorable to cultivateing 11 internal portions's of chamber cell breathing on the one hand, and on the other hand also makes things convenient for container lid 1 can be sealed completely to avoid because need ventilate for cultivateing the chamber 11 after the centrifugation and cause the pollution to the culture medium. The surface of the container mouth 3 is fixedly provided with an anti-overflow flange 14, so that the sealing efficiency of the container mouth 3 in the centrifugal process is improved. The container cover 1 is a self-sealing container cover 1, and sealing efficiency is improved.
Wherein, the upper closing plate 12 and the lower closing plate 13 are both concave closing plates. The concave design of the upper sealing plate 12 helps the culture medium solution stained on the inner wall of the upper sealing plate 12 to concentrate along the inclined direction and finally fall into the culture cavity 11 after centrifugation. The concave design of the lower closure plate 13 helps the pipette to completely withdraw the medium at the concave point when it is desired to pipette or remove the medium. The pits are the middle portion of the front view. The pipette is facilitated to find the lowest liquid level in the culture chamber 11. The legs 7 and the partition are intermittently distributed in a front view. Facilitating better gripping means of the robotic arm.
In addition, the container cover 1, the self-sealing container cover 1, is liquid-tight during centrifugation or culture; the outer shell 9, the bottom surface is a smooth polished plane, guarantee the effective combination of cell and gene vector virus in the course of centrifuging; the waterproof breathable film 10 is suitable for culturing 20-100 ml of cell solution in a bottle and provides efficient exchange of carbon dioxide and oxygen in the incubator; the rib plate 6 is convenient for clamping and releasing the robot clamping jaw after being inserted; an intra-cavity ramp. The intracavity slope mainly is convenient for move liquid syringe needle and can follow the bottleneck and go deep into the bottle end after, can absorb clean with the cell sap well, reduces liquid and remains.
It should be pointed out that the pressing shaft 6 mentioned in the present invention does not affect the feasibility of the solution, whether it is rotating or not. And is not intended to be limiting.
In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (9)
1. The utility model provides a centrifugal transfection and co-culture integration consumptive material that is fit for automated production, includes the shell body, goes up the shrouding and down the shrouding, its characterized in that: the upper sealing plate is fixedly connected to the upper end surface of the outer shell through a convex shoulder, and the two sides of the upper sealing plate are fixedly provided with outer sealing plates; the upper sealing plate is provided with a container opening, and a container cover is connected to the container opening in a threaded manner; the outer shrouding lower extreme is provided with down the shrouding, lower shrouding lower extreme evenly is provided with a plurality of gusset, the fixed a plurality of stabilizer blades that are provided with of shell body lower extreme.
2. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 1, characterized in that: the outer sealing plate is provided with a window.
3. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 2, characterized in that: the windows are distributed in an up-and-down symmetrical mode, and the number of the windows is more than one.
4. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 3, characterized in that: and a waterproof breathable film is fixedly arranged in the window.
5. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 1, characterized in that: the surface of the container mouth is fixedly provided with an anti-overflow flange.
6. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 1, characterized in that: the container cover is a self-sealing container cover.
7. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 1, characterized in that: the upper sealing plate and the lower sealing plate are both concave sealing plates.
8. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 7, characterized in that: the pits are the middle portion of the front view.
9. The integrated consumable for centrifugal transfection and co-culture suitable for automated production according to claim 1, characterized in that: the support legs and the rib plates are intermittently distributed in the front view direction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010198625.9A CN111378559A (en) | 2020-03-20 | 2020-03-20 | Centrifugal transfection and combined culture integrated consumable suitable for automatic production |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010198625.9A CN111378559A (en) | 2020-03-20 | 2020-03-20 | Centrifugal transfection and combined culture integrated consumable suitable for automatic production |
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| CN111378559A true CN111378559A (en) | 2020-07-07 |
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| CN202010198625.9A Pending CN111378559A (en) | 2020-03-20 | 2020-03-20 | Centrifugal transfection and combined culture integrated consumable suitable for automatic production |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004105139A (en) * | 2002-09-20 | 2004-04-08 | Sanyo Electric Co Ltd | Method for centrifugal separation of cell and centrifugal separation apparatus therefor |
| WO2011130865A2 (en) * | 2010-04-21 | 2011-10-27 | Yves Larcher | Automated cell culture system |
| CN107904173A (en) * | 2017-12-27 | 2018-04-13 | 深圳北科赛动生物机器人有限公司 | Biological culture flask |
| CN108060081A (en) * | 2018-02-05 | 2018-05-22 | 中山大学 | A kind of multi-layer planar filling type bioreactor |
| CN108102914A (en) * | 2017-12-27 | 2018-06-01 | 深圳北科赛动生物机器人有限公司 | Stem cell auto culturing system |
| CN208717353U (en) * | 2018-05-25 | 2019-04-09 | 广州杰特伟生物科技有限公司 | Tissue Culture Flask |
-
2020
- 2020-03-20 CN CN202010198625.9A patent/CN111378559A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004105139A (en) * | 2002-09-20 | 2004-04-08 | Sanyo Electric Co Ltd | Method for centrifugal separation of cell and centrifugal separation apparatus therefor |
| WO2011130865A2 (en) * | 2010-04-21 | 2011-10-27 | Yves Larcher | Automated cell culture system |
| CN107904173A (en) * | 2017-12-27 | 2018-04-13 | 深圳北科赛动生物机器人有限公司 | Biological culture flask |
| CN108102914A (en) * | 2017-12-27 | 2018-06-01 | 深圳北科赛动生物机器人有限公司 | Stem cell auto culturing system |
| CN108060081A (en) * | 2018-02-05 | 2018-05-22 | 中山大学 | A kind of multi-layer planar filling type bioreactor |
| CN208717353U (en) * | 2018-05-25 | 2019-04-09 | 广州杰特伟生物科技有限公司 | Tissue Culture Flask |
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Application publication date: 20200707 |