CN111826287A - Improved cell culture perfusion device - Google Patents
Improved cell culture perfusion device Download PDFInfo
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- CN111826287A CN111826287A CN202010773673.6A CN202010773673A CN111826287A CN 111826287 A CN111826287 A CN 111826287A CN 202010773673 A CN202010773673 A CN 202010773673A CN 111826287 A CN111826287 A CN 111826287A
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- bottle
- perfusion
- pipe
- cell culture
- improved cell
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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
- C12M23/52—Mobile; Means for transporting the apparatus
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B9/00—Cleaning hollow articles by methods or apparatus specially adapted thereto
- B08B9/08—Cleaning containers, e.g. tanks
- B08B9/087—Cleaning containers, e.g. tanks by methods involving the use of tools, e.g. brushes, scrapers
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
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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
- C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
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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
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/02—Means for pre-treatment of biological substances by mechanical forces; Stirring; Trituration; Comminuting
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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
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/20—Heating; Cooling
Abstract
The invention discloses an improved cell culture perfusion device, which comprises a perfusion bottle, wherein the opening of the perfusion bottle is downward arranged, the bottom of the perfusion bottle is provided with a hopper, the lower end of the hopper is communicated with a discharge pipe, the bottom of the outer side wall of the perfusion bottle is connected with a vertical frame, the left side of the top of the vertical frame is connected with a storage box, a feed pipe is communicated between the storage box and the top of the perfusion bottle, the top of the right side wall of the perfusion bottle is communicated with a gas injection pipe, the upper part of the inner cavity of the perfusion bottle is provided with a cleaning mechanism, and the center of the perfusion bottle is provided with a cut-off mechanism; the invention has reasonable structural design, is beneficial to controlling the cell perfusion quantity, ensures the culture efficiency, can reduce the invasion of external impurities, is convenient to disassemble and is beneficial to cleaning and maintaining the components.
Description
Technical Field
The invention relates to the technical field of cell culture related devices, in particular to an improved cell culture perfusion device.
Background
With the wide application of biotechnology drugs in diagnosis, treatment and prevention of human diseases, the industrialization process of large-scale high-efficiency culture of animal cell expression products is greatly accelerated. The large-scale culture of animal cells has become one of the most important key technologies in the field of biopharmaceuticals, and the research and development thereof have promoted the rapid development of the biotechnology industry. In recent years, the market availability and the huge economic benefits brought by protein drugs have raised the hot tide of mammalian cell culture. How to perfect the culture process of animal cells with long multiplication time, sensitivity to external environment and high culture difficulty, improve the expression quantity of cell proteins, and effectively put into large-scale production becomes a hot point of domestic and foreign research.
Perfusion culture technology is a common fermentation technology, and mostly adopts a stirring type cell fermentation system and also can adopt a tubular system. During the fermentation, the culture solution is continuously perfused into the fermentation tank and flows out at the same flow rate. The perfusion process has the advantages that cell debris and byproducts can be continuously removed in the fermentation process, and the influence of various enzymes released after cell death on products can be reduced. Meanwhile, a relatively constant culture environment is provided by continuously perfusing fresh culture solution, so that a relatively high yield per unit volume can be obtained. As the perfusion system mostly adopts a cell interception and recovery system, the high-density fermentation of cells can be realized.
At present, the selection of animal cell culture methods is mainly determined by the characteristics of cell growth and the properties of the target protein. The perfusion culture has the obvious advantages that the protein can be separated out in time along with the culture medium, the retention time of the protein in the reactor is short, the protein is less degraded by various hydrolytic enzymes in a culture system, and the quality of the protein is improved. This feature is very advantageous for the production of chemically unstable proteins, such as enzymes, coagulation factors, etc. In the prior art, a cell culture perfusion device is similar to a funnel, the perfusion quantity of cells cannot be controlled, the culture efficiency is influenced, meanwhile, the perfusion device lacks a cleaning function, and cross contamination is easy to generate.
Disclosure of Invention
The present invention aims to overcome the above problems in the prior art and to provide an improved cell culture perfusion apparatus. In order to achieve the technical purpose and achieve the technical effect, the invention is realized by the following technical scheme:
the utility model provides an improved generation cell culture perfusion device, includes the perfusion bottle, perfusion bottle opening sets up downwards, the hopper is installed to the bottom of perfusion bottle, hopper lower extreme intercommunication has the discharging pipe, the lateral wall bottom of perfusion bottle is connected with the grudging post, the top left side of grudging post is connected with the storage case, the intercommunication has the inlet pipe between storage case and the perfusion bottle top, the right side wall top intercommunication of perfusion bottle has the gas injection pipe, the inner chamber upper portion of perfusion bottle is equipped with clean mechanism, the center department of perfusion bottle is equipped with the mechanism that dams.
Preferably, among an improved generation cell culture perfusion device, the mechanism that dams includes the montant, the lower extreme of montant is connected with the toper stopper, sealing bearing is installed on the circumference lateral wall upper portion of montant, sealing bearing's outer wall connection has the limiting plate, be connected with the folding tube between the inner chamber top of limiting plate and perfusion bottle, the upper end of montant is equipped with the external screw thread, perfusion bottle top is seted up with external screw thread matched with screw hole, the outside cover of montant is equipped with reset spring, reset spring is located the folding tube.
Preferably, in an improved generation cell culture perfusion device, clean mechanism includes the sealed shell at two sets of intercommunication perfusion bottle tops, and is two sets of sealed shell bilateral symmetry sets up, and is two sets of the inner chamber top of sealed shell all is connected with electric putter, the one end that electric putter stretches into the perfusion bottle inner chamber is connected with the annular plate, the circumference lateral wall of annular plate is connected with the scraper blade, the inner chamber lateral wall of scraper blade laminating perfusion bottle.
Preferably, in an improved generation cell culture perfusion device, the gas injection pipe is including the L venturi tube of intercommunication perfusion bottle, L venturi tube opening sets up downwards, the vertical end inner chamber of L venturi tube is equipped with the piston, the bottom of piston is connected with the push rod, the air inlet has been seted up to L venturi tube vertical end.
Preferably, in an improved generation cell culture perfusion device, the inner chamber bottom of storage case is connected with the heating pipe, the left side wall of storage case is connected with driving motor, driving motor's right side power take off end is connected with the (mixing) shaft, the one end that the (mixing) shaft stretched into the storage case inner chamber is connected with the stirring leaf.
Preferably, in the improved cell culture perfusion device, the feed pipe is provided with a liquid pump, and one end of the feed pipe extending into the perfusion bottle is positioned above the opening of the annular plate.
Preferably, in the improved cell culture perfusion device, the upper part of the hopper is connected with the lower end of the perfusion bottle in a threaded manner, and the bottom of the perfusion bottle is connected with a sealing ring.
Preferably, in the improved cell culture perfusion device, the conical plug is positioned right above the discharge pipe, and the diameter of the upper end of the conical plug is larger than the inner diameter of the discharge pipe.
The invention has the beneficial effects that:
the cell perfusion device has reasonable structural design, the culture solution is stored in the storage box, the culture solution is sent into the perfusion bottle along the feeding pipe by the liquid pump, and the relative position of the interception mechanism and the discharge pipe can be adjusted by matching the vertical rod, the external thread, the threaded hole, the reset spring and the limiting plate, so that the interception operation of the culture solution is completed, the cell perfusion quantity can be controlled, and the culture efficiency is guaranteed; move up through the push rod drive piston, can accomplish the notes oxygen operation of culture solution, the gas injection pipe opening sets up downwards, can reduce external impurity's invasion, and the perfusion finishes the back, drives the annular slab through electric putter and descends, and the scraper blade can clear up perfusion bottle inner wall to prevent remaining adhering to, hopper and perfusion bottle adoption spiro union installation are convenient for dismantle, are favorable to the clearance maintenance of subassembly.
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 external view of the present invention;
FIG. 2 is a schematic view of the internal structure of the present invention;
FIG. 3 is a schematic structural view of the shut-off mechanism of the present invention;
FIG. 4 is a schematic view of the cleaning mechanism of the present invention;
FIG. 5 is a schematic view showing the structure of a gas injection tube according to the present invention;
FIG. 6 is a schematic view of the storage bin of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings 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 of the 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. 1-6, the present embodiment is an improved cell culture perfusion apparatus, including a perfusion bottle 1, wherein the perfusion bottle 1 has a downward opening, a hopper 2 is installed at the bottom of the perfusion bottle 1, a discharging pipe 3 is communicated with the lower end of the hopper 2, a vertical frame 4 is connected to the bottom of the outer side wall of the perfusion bottle 1, a storage tank 5 is connected to the left side of the top of the vertical frame 4, a feeding pipe 6 is communicated between the storage tank 5 and the top of the perfusion bottle 1, a gas injection pipe 7 is communicated with the top of the right side wall of the perfusion bottle 1, a cleaning mechanism 8 is disposed on the upper portion of the inner cavity of the perfusion bottle 1, and a shut-off mechanism 9 is disposed at the center of the perfusion bottle.
The closure mechanism 9 includes montant 901, the lower extreme of montant 901 is connected with toper stopper 902, sealed bearing 903 is installed on the circumference lateral wall upper portion of montant 901, sealed bearing 903's outer wall connection has limiting plate 904, be connected with folding tube 905 between limiting plate 904 and perfusion bottle 1's the inner chamber top, the upper end of montant 901 is equipped with external screw thread 906, perfusion bottle 1 top is seted up with external screw thread 906 matched with screw hole, the outside cover of montant 901 is equipped with reset spring 907, reset spring 907 is located folding tube 905.
The cleaning mechanism 8 comprises two groups of sealing shells 801 communicated with the top of the perfusion bottle 1, the two groups of sealing shells 801 are arranged in a bilateral symmetry mode, the tops of inner cavities of the two groups of sealing shells 801 are connected with electric push rods 802, one end, extending into the inner cavity of the perfusion bottle 1, of each electric push rod 802 is connected with an annular plate 803, the outer circumferential side wall of each annular plate 803 is connected with a scraping plate 804, and the scraping plates 804 are attached to the inner cavity side wall of the perfusion bottle 1.
The gas injection pipe 7 comprises an L-shaped pipe 701 communicated with the perfusion bottle 1, an opening of the L-shaped pipe 701 is arranged downwards, a piston 702 is arranged in an inner cavity of a vertical end of the L-shaped pipe 701, a push rod 703 is connected to the bottom of the piston 702, and a gas inlet hole 704 is formed in the vertical end of the L-shaped pipe 701.
The feed pipe 6 is provided with a liquid pump 601, and one end of the feed pipe 6 extending into the perfusion bottle 1 is positioned above the opening of the annular plate 803, so that the culture solution can be conveniently injected.
The upper part of the hopper 2 is connected with the lower end of the perfusion bottle 1 in a screw connection mode, and the bottom of the perfusion bottle 1 is connected with a sealing ring, so that the loading and unloading are convenient.
One specific application of this embodiment is:
when the device works, the vertical frame 4 is moved to a working place and is connected with an external power supply, the discharge pipe 3 is arranged at the upper end of a culture dish and is stored with culture solution through the storage box 5, the culture solution is heated through the heating pipe 501, the stirring shaft 503 is driven to rotate through the driving motor 502, the culture solution is uniformly stirred by the stirring blades 504, the culture solution is pumped into the feed pipe 6 through the liquid pumping pump 601 and is further fed into the perfusion bottle 1, the vertical rod 901 is rotated, the external thread 906 is separated from the threaded hole, the limiting plate 904 is pushed by the elastic force of the reset spring 907 to drive the vertical rod 901 to descend, the end part of the discharge pipe 3 can be attached and sealed through the conical plug 902, so that the interception operation of the culture solution is completed, the sealing performance is improved through the folding pipe 905 and the sealing bearing 903, the invasion of external impurities is reduced, the piston 702 is driven to move upwards through the push rod 703, can accomplish the notes oxygen operation of culture solution, 7 openings of gas injection pipe set up downwards, can reduce the invasion of external impurity, the culture solution can flow out along discharging pipe 3 when upwards pulling montant 901, the perfusion finishes the back, drive annular plate 803 through electric putter 802 and descend, scraper blade 804 can clear up perfusion bottle 1 inner wall, thereby prevent remaining adhering to, hopper 2 adopts the spiro union installation with perfusion bottle 1, be convenient for dismantle, be favorable to the clearance maintenance of subassembly.
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 (8)
1. An improved cell culture perfusion device comprises a perfusion bottle (1), and is characterized in that: irritate and flow bottle (1) opening and set up downwards, hopper (2) are installed to the bottom of irritating bottle (1), hopper (2) lower extreme intercommunication has discharging pipe (3), the lateral wall bottom of irritating bottle (1) is connected with grudging post (4), the top left side of grudging post (4) is connected with storage case (5), the intercommunication has inlet pipe (6) between storage case (5) and the irritating bottle (1) top, the right side wall top intercommunication of irritating bottle (1) has gas injection pipe (7), the inner chamber upper portion of irritating bottle (1) is equipped with clean mechanism (8), the center department of irritating bottle (1) is equipped with cut-off mechanism (9).
2. An improved cell culture perfusion apparatus as claimed in claim 1, wherein: the cutoff mechanism (9) comprises a vertical rod (901), the lower end of the vertical rod (901) is connected with a conical plug (902), a sealing bearing (903) is installed on the upper portion of the outer side wall of the circumference of the vertical rod (901), the outer wall of the sealing bearing (903) is connected with a limiting plate (904), a folding pipe (905) is connected between the limiting plate (904) and the top of an inner cavity of the perfusion bottle (1), an external thread (906) is arranged at the upper end of the vertical rod (901), a threaded hole matched with the external thread (906) is formed in the top of the perfusion bottle (1), a reset spring (907) is sleeved outside the vertical rod (901), and the reset spring (907) is located in the folding pipe (905).
3. An improved cell culture perfusion apparatus as claimed in claim 1, wherein: clean mechanism (8) are including sealed shell (801) at two sets of intercommunication perfusion bottle (1) tops, and are two sets of sealed shell (801) bilateral symmetry sets up, and is two sets of the inner chamber top of sealed shell (801) all is connected with electric putter (802), the one end that electric putter (802) stretched into perfusion bottle (1) inner chamber is connected with annular plate (803), the circumference lateral wall of annular plate (803) is connected with scraper blade (804), the inner chamber lateral wall of scraper blade (804) laminating perfusion bottle (1).
4. An improved cell culture perfusion apparatus as claimed in claim 1, wherein: the gas injection pipe (7) comprises an L-shaped pipe (701) communicated with the perfusion bottle (1), an opening of the L-shaped pipe (701) is arranged downwards, a piston (702) is arranged in an inner cavity of a vertical end of the L-shaped pipe (701), a push rod (703) is connected to the bottom of the piston (702), and an air inlet hole (704) is formed in the vertical end of the L-shaped pipe (701).
5. An improved cell culture perfusion apparatus as claimed in claim 1, wherein: the heating pipe (501) is connected to the inner chamber bottom of storage case (5), the left side wall of storage case (5) is connected with driving motor (502), the right side power take off end of driving motor (502) is connected with (mixing) shaft (503), the one end that (mixing) shaft (503) stretched into storage case (5) inner chamber is connected with stirring leaf (504).
6. An improved cell culture perfusion apparatus as claimed in claim 3, wherein: the liquid pump (601) is installed on the feeding pipe (6), and one end, extending into the filling bottle (1), of the feeding pipe (6) is located above an opening of the annular plate (803).
7. An improved cell culture perfusion apparatus as claimed in claim 1, wherein: the upper part of the hopper (2) is connected with the lower end of the perfusion bottle (1) in a threaded connection mode, and the bottom of the perfusion bottle (1) is connected with a sealing ring.
8. An improved cell culture perfusion apparatus as claimed in claim 2, wherein: the conical plug (902) is positioned right above the discharge pipe (3), and the diameter of the upper end of the conical plug (902) is larger than the inner diameter of the discharge pipe (3).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010773673.6A CN111826287A (en) | 2020-08-04 | 2020-08-04 | Improved cell culture perfusion device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010773673.6A CN111826287A (en) | 2020-08-04 | 2020-08-04 | Improved cell culture perfusion device |
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| CN111826287A true CN111826287A (en) | 2020-10-27 |
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| CN202010773673.6A Pending CN111826287A (en) | 2020-08-04 | 2020-08-04 | Improved cell culture perfusion device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112877210A (en) * | 2021-02-25 | 2021-06-01 | 孔祥玲 | Stem cell culture perfusion device with controllable pH value |
| CN113897278A (en) * | 2021-09-26 | 2022-01-07 | 李同辉 | Nutrient solution adding equipment used in biological cultivation |
| CN114159223A (en) * | 2021-11-10 | 2022-03-11 | 南京纳中安商贸有限公司 | Ophthalmic dropping device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN207418758U (en) * | 2017-10-26 | 2018-05-29 | 天津百恩生物科技有限公司 | A kind of stem cell culture perfusing device |
| CN208857308U (en) * | 2018-08-09 | 2019-05-14 | 卡替（上海）生物技术股份有限公司 | Stem cell culture perfusing device |
| CN209685811U (en) * | 2019-03-07 | 2019-11-26 | 山东大学 | A kind of cell culture gaseous environment regulating device |
-
2020
- 2020-08-04 CN CN202010773673.6A patent/CN111826287A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN207418758U (en) * | 2017-10-26 | 2018-05-29 | 天津百恩生物科技有限公司 | A kind of stem cell culture perfusing device |
| CN208857308U (en) * | 2018-08-09 | 2019-05-14 | 卡替（上海）生物技术股份有限公司 | Stem cell culture perfusing device |
| CN209685811U (en) * | 2019-03-07 | 2019-11-26 | 山东大学 | A kind of cell culture gaseous environment regulating device |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112877210A (en) * | 2021-02-25 | 2021-06-01 | 孔祥玲 | Stem cell culture perfusion device with controllable pH value |
| CN112877210B (en) * | 2021-02-25 | 2022-07-26 | 上海揽微赛尔生物科技有限公司 | Stem cell culture perfusion device with controllable pH value |
| CN113897278A (en) * | 2021-09-26 | 2022-01-07 | 李同辉 | Nutrient solution adding equipment used in biological cultivation |
| CN114159223A (en) * | 2021-11-10 | 2022-03-11 | 南京纳中安商贸有限公司 | Ophthalmic dropping device |
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