CN101899394B - External circulation animal cell culture bioreactor - Google Patents
External circulation animal cell culture bioreactor Download PDFInfo
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- CN101899394B CN101899394B CN 201010242025 CN201010242025A CN101899394B CN 101899394 B CN101899394 B CN 101899394B CN 201010242025 CN201010242025 CN 201010242025 CN 201010242025 A CN201010242025 A CN 201010242025A CN 101899394 B CN101899394 B CN 101899394B
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- 210000004102 animal cell Anatomy 0.000 title claims abstract description 21
- 238000004113 cell culture Methods 0.000 title claims abstract description 21
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 30
- 239000007788 liquid Substances 0.000 claims description 30
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 25
- 239000001301 oxygen Substances 0.000 claims description 21
- 229910052760 oxygen Inorganic materials 0.000 claims description 21
- 239000000523 sample Substances 0.000 claims description 16
- 238000012544 monitoring process Methods 0.000 claims description 13
- 239000007789 gas Substances 0.000 claims description 12
- 238000005520 cutting process Methods 0.000 claims description 8
- 229910001220 stainless steel Inorganic materials 0.000 claims description 8
- 239000011229 interlayer Substances 0.000 claims description 7
- 239000010935 stainless steel Substances 0.000 claims description 7
- 230000001954 sterilising effect Effects 0.000 abstract description 15
- 238000000034 method Methods 0.000 abstract description 13
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 11
- 238000004114 suspension culture Methods 0.000 abstract description 4
- 230000002349 favourable effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 27
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 230000008569 process Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 241000700605 Viruses Species 0.000 description 5
- 239000003513 alkali Substances 0.000 description 5
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- 238000003756 stirring Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
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- 238000004519 manufacturing process Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
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- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229910001882 dioxygen Inorganic materials 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
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- 238000011017 operating method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
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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
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to an external circulation animal cell culture bioreactor which comprises a tank body, an external circulation pipeline, a heater, a peristaltic pump and a control system, wherein the tank body comprises a large tank and a small tank; the small tank is sheathed in the large tank and is clamped and fixed with the large tank respectively through ducts D, A and G; the external circulation pipeline comprises a tank external circulation pipeline and ducts A, B, E and D; on the upper part of the outside of the tank body, the ducts A and B are connected with the tank external circulation pipeline through a tee joint; on the lower part of the outside of the tank body, the ducts E and D are connected with the tank external circulation pipeline through a tee joint; the tank external circulation pipeline penetrates through the peristaltic pump; and the lowest position of the duct D is connected with a drainage pipe. The invention fully displays the characteristics of the animal cell large-scale culture technique, has the characteristics of thorough sterilization, simple operation, low pollution rate of cell culture, good quality and high yield, has favorable functionality and economical efficiency, and can be widely used for animal cell microcarrier suspension culture and animal cell suspension culture.
Description
Technical field
The present invention relates to adopt suspension process to carry out the device of animal cell culture, relate in particular to a kind of external circulation animal cell culture bioreactor.
Background technology
Bio-reactor refers to viable cell or enzyme to be that biological catalyst carries out the equipment that cell proliferation or biochemical reaction provide control environment, and it is the key equipment in the bioprocesses.Selected quality, yield (transformation efficiency) and the energy consumption to the biochemical industry product of the structure of bio-reactor, operating method and operational condition has substantial connection.
Along with the continuous development of biotechnology, extensive zooblast bioreactor culture technology just progressively replaces vial and leaves standstill cultivation and rotating and culturing technology, because it provide the living environment of the best can in time cell and virus; Thereby the output and the quality of the raising product of top efficiency; Should this be a qualitative leap on the animal cell culture history, also be the important breakthrough of medicine biological technique, and it has not only simplified technology; The more important thing is that it can produce high-quality product; A variety of bio-reactors are arranged in the market, comprise respectively having superiority home-made and import and defective; That have even have inevitable hard defects, thus limited the maximum performance of its advantage.
To compare analysis with regard to the design of aspects such as alr mode, supervisory system, sterilization method, ventilating mode, blowdown pattern to existing several kinds of animal cell culture bioreactors below, please see following table for details:
Summary of the invention
In view of existing existing the problems referred to above of animal cell culture bioreactor; And for the quality of cell cultures, the influence that yield caused; The present invention is intended to disclose a kind of external circulation animal cell culture bioreactor; With the effect of effective assurance cell cultures, embody good and economic and functional.
Technical solution of the present invention is achieved in that
A kind of external circulation animal cell culture bioreactor comprises tank body and external circulation line, well heater, peristaltic pump and system, it is characterized in that:
Said tank body comprises big jar and canister; Said big jar comprises big tank, water-bath interlayer and cover, and said cover is provided with ventage; Said canister comprises lower cover and middle tank body, and the said lower cover of going up is tightly connected with middle tank body respectively; Through being arranged at lower cover and middle tank body junction on the said canister inside and perpendicular to the web plate of stainless steel up and down of the axis of said canister, following three spaces during said canister is divided into;
Said canister is placed on said big jar inside; And fix through pipeline D, A, G and said big jar of cutting ferrule respectively; That is: the interior extreme lower position of lower cover of canister is fixed and be communicated to pipeline D one end; The bottom that its other end is arrogant jar is connected to outside big jar and through threeway and is connected with the I section and the pipeline E of jar external circulation line respectively, and the arrogant tank body of the other end of said pipeline E bottom is inserted in big jar; The extreme higher position that pipeline A one end is fixed in the upper cover of canister also is communicated in the canister since then; The top of the arrogant tank wall of its other end picks out outside the tank body; And be communicated with the II section and the pipeline B of jar external circulation line through threeway; Pipeline B is equipped with diaphragm valve in this port, and the arrogant tank wall of the other end of said pipeline B middle and upper part is connected in big jar; The end of pipeline G is fixed in the upper cover of canister and passes the upper cover of canister and the middle tank body that last web plate is connected to said canister successively, and its other end is switched to outside the big tank;
Said external circulation line comprises pipeline E and the pipeline D that pipeline A that the outer top of a jar external circulation line, tank body is connected with the II section of said jar of external circulation line through threeway and pipeline B, the outer bottom of tank body are connected with the I section of said jar of external circulation line through threeway; Said jar of external circulation line passes said peristaltic pump; Said peristaltic pump adopts electrodeless variable-speed;
Also connect liquid respectively on the said jar of external circulation line and replenished pipeline, gas make-up pipeline, liquid collecting tube road;
Said pipeline D connects blow-off pipe at its lowest order.
Further; The monitoring probe that has also connected pH control and dissolved oxygen control on the II section of said jar of external circulation line; Be placed with temperature controlled monitoring probe in the said big tank; Above-mentioned monitoring probe is connected with housing through lead, and said housing comprises indicating meter, dissolved oxygen unit, pH unit, temperature regulator, datalogger.
Further, the arrogant tank body of said pipeline E bottom is inserted in big jar along big tank wall tangential.
Further, the said lower cover of going up is tightly connected through external clip with middle tank body respectively.
Compared with prior art, the design of external circulation animal cell culture bioreactor of the present invention has demonstrated fully the characteristics of animal cell large-scale culture technique, and layout of beam line is succinct; Have excellent function property, safety, accessibility, the existing specific descriptions as follows:
(1) alr mode and sealing: utilize liquid circulation from bottom to top that nutrient solution and cell are stirred through peristaltic pump, fully airtight loop makes the PH, temperature, dissolved oxygen etc. of liquid reach full and uniform, and the shearing force of pair cell goes to zero; Solved the problem of the stirring of other form simultaneously to the seal request of tank body;
(2) ventilating mode: deep layer is directly ventilated, bubble fully not with cells contacting, thereby need not the pin bulb apparatus, can avoid the injury of bubble fully for cell;
(3) sterilization method: reasonable in design, layout of beam line is succinct, in the moist heat sterilization cabinet, sterilizes, and is simple to operate, and the dead angle of not sterilizing also need not the maintenance management of pressing force container, has more security; Simultaneously shortened the production preparatory period greatly;
(4) blowdown pattern: can emit at any time through the discharge pipe that is in least significant end come off, apoptotic cells and broken microcarrier, reduced pollution rate;
Monitoring such as (5) supervisory system: PH, temperature and dissolved oxygen probe is installed on the outer circulation loop, and because of outer circulation makes liquid mixing full and uniform, so monitoring data is more objective, accurate and more representative.
To sum up, scientific structure design of the present invention, layout of beam line is succinct; Cell and nutrient solution are isolated cleverly, and best living environment is provided in time for cell and virus, evaded many hard defects and the defective that exists the existing installation from structure; Give full play to the characteristics of animal cell large-scale culture technique, had sterilization thoroughly, simple to operate; The pollution of cell culture rate is low, and quality is good, the distinguishing feature that yield is high; Have excellent function property, economy, can be adaptable across zooblast microcarrier suspension culture and zooblast suspension culture.
Description of drawings
Fig. 1 is the structural representation of the embodiment of the invention;
Fig. 2 is the vertical view of Fig. 1.
Embodiment
Combine accompanying drawing at present, the present invention is made further specific descriptions.
Said external circulation animal cell culture bioreactor; Comprise tank body and external circulation line, well heater, peristaltic pump and system; Wherein well heater, peristaltic pump and system adopt existing molding device; Tank structure and external circulation line have then been done bigger innovation, and be as depicted in figs. 1 and 2:
Said tank body comprises big jar 1 and canister 2; Said big jar 1 comprises big tank 11, water-bath interlayer 12 and cover 13; One side lower part of said water-bath interlayer 12 is provided with hot water tube for entering can road F; The top of offside is provided with hot water and goes out a jar pipeline C; Said hot water tube for entering can road F goes out jar pipeline C with hot water and is connected with the circulation line of well heater respectively, and said cover 13 is provided with ventage and connects breather line H, K; Said canister 2 comprises lower cover 21,23 and middle tank body 22, through external clip 24,25 the said lower cover of going up is tightly connected with middle tank body 22 respectively; Said canister 2 is inner goes up lower covers and middle tank body junction and perpendicular to the web plate of stainless steel up and down 26,27 of the axis of said canister through being arranged at, following three spaces during said canister 2 is divided into;
Said canister 2 is placed on said big jar 1 inside; And fix through pipeline D, A, G and said big jar of 2 cutting ferrules respectively; That is: pipeline D one end fixes and is communicated to the extreme lower position in the lower cover 23 of canister through cutting ferrule; The bottom that its other end is arrogant jar 1 is connected to outside big jar and through threeway and is connected with the I section and the pipeline E of jar external circulation line respectively, and the arrogant tank body of the other end of said pipeline E bottom is inserted in the big jar 1 along the tangential of big tank wall; Pipeline A one end through cutting ferrule fixing with and be communicated in the canister 2 from the extreme higher position of the upper cover 21 of canister; The top of the arrogant tank wall of its other end picks out outside the tank body; And be communicated with the II section and the pipeline B of jar external circulation line through threeway; Pipeline B is equipped with diaphragm valve 6 in this port, and the arrogant tank wall of the other end of said pipeline B middle and upper part is connected in big jar 1; The end of pipeline G through cutting ferrule fixedly upper cover 21 and the upper cover 21 that passes canister successively and the last web plate 26 of canister be connected to the middle tank body 22 of said canister, its other end is switched to outside the tank body;
Said external circulation line comprises pipeline E and the pipeline D that pipeline A that the outer top of a jar external circulation line, tank body is connected with said jar of external circulation line through threeway and pipeline B, the outer bottom of tank body are connected with said jar of external circulation line through threeway; Said jar of external circulation line passes said peristaltic pump;
The said jar of external circulation line that is connected through threeway with pipeline A and pipeline B before the aforementioned entering peristaltic pump is referred to as the II section of jar external circulation line; The said jar of external circulation line that another section promptly comes out with pipeline E and pipeline D are connected through threeway of correspondence is referred to as the I section of jar external circulation line in said peristaltic pump; Automatic control probe, the additional pipeline 71 of liquid, gas make-up pipeline 72, liquid collecting tube road 73 have also been connected on the II section of said jar of external circulation line respectively; Said automatic control probe comprises the monitoring probe 31 of pH control and the monitoring probe 32 of dissolved oxygen control; Be placed with temperature controlled monitoring probe 33 in the big tank, above-mentioned monitoring probe is connected with housing through lead.
Said pipeline D connects blow-off pipe 5 at its lowest order.
The material of said tank body is the 316L stainless steel, and said water-bath interlayer is selected 304 stainless steels for use, the band thermal insulation layer.The tank body top is equipped with lamp mirror and visor, is used for inside reactor and observes.
When carrying out the tank body assembling; The last lower cover 21,23 of said canister connects with clip 24,25 respectively with middle tank body 22, catches up with down two end sockets 21,23 to middle tank body 22 through stainless steel web plate 26,27 and keeps apart, and lower cover 23 connects with pipeline D through the mode of cutting ferrule; Upper cover connects with pipeline A, G through cutting ferrule; After built-in canister 2 fixes, cover big jar cover 13, whole tank body assembled.
The connection of tank body pipeline comprises that pipeline C, F are connected to form big jar heating in water bath loop respectively with well heater; Pipeline A, B (connecting diaphragm valve 6) pass through the II section that the stainless steel threeway connects jar external circulation line, also insert liquid on the II section of said pipeline respectively and replenish pipeline, gas make-up pipeline, liquid collecting tube road and comprise the monitoring probe that pH control and dissolved oxygen are controlled; Pipeline E, D connect the I section of jar external circulation line through the stainless steel threeway; Said jar of external circulation line passes said peristaltic pump; Pipeline H, K are connected on the ventage of big jar cover 13, and pipeline G is that built-in canister 2 connects the outside passages of tank bodies.
Sterilising treatment:
After above-mentioned tank body assembling and pipe connection are accomplished; Directly be put into whole big jar that to carry out sterilising treatment in the moist heat sterilization cabinet subsequent use; So just need not do the checking of very complicated on-line cleaning and on-line cleaning; The more important thing is not stay the sterilization dead angle, thereby greatly reduce pollution rate, also do not need the pressing force container to come use and management because of online sterilization.About 1 day of above-mentioned sterilising treatment link required time, simple to operate, sterilization is thoroughly; And on average about 7 days of the online vapor sterilization processing links required time of existing installation, complicated operation stays the sterilization dead angle easily, pollutes, and must the maintenance management of pressing force container.
Stir control:
Realize the liquid circulation stirring through peristaltic pump.Microcarrier and cell are sent into through pipeline G in the middle tank body 22 of canister, and through web plate 26,27 liquid of microcarrier and cell and big jar are isolated; Under the effect of peristaltic pump, the liquid in the canister is mobile from bottom to top, and the liquid in the big jar rotates and produces eddy current, realizes that liquid fully stirs, and parameters such as fluid temperature, pH value, dissolved oxygen are in full accord, and recycle pump adopts electrodeless variable-speed, operates steadily.
System:
The reactor drum function is realized by housing, comprises image display, dissolved oxygen unit, PH unit, temperature regulator, datalogger.Wherein,
Temperature control:
Adopt the water-bath interlayer that reactor drum is carried out temperature control.The water-bath interlayer is equipped with water-bath heater, and water-bath heater has improved the susceptibility and the stability of temperature of reactor control.Reactor temperature control system adopts the pid control mode based on the tank body temperature, and temperature compensation function makes temperature control more stable, temperature control precision ± 0.3 ℃.
PH control:
Adopt the standalone module mode, the pH unit is provided with separately.Use the PID master mode to realize control automatically, have Adaptive PID Control adjustment function, the pid parameter that automatically setting is best.PH control and carbonic acid gas breather valve, and acid, alkali peristaltic pump is associated, and through the switch of control carbonic acid gas, acid solution, alkali lye, realizes the automatic control of pH.When pH was lower than set(ting)value, unit started the alkali lye peristaltic pump, mended alkali and improved pH to set(ting)value; When pH was higher than set(ting)value, unit started the dioxide gas SV, and the acid solution peristaltic pump, reduced pH to set(ting)value.Carbon dioxide flow can be regulated through spinner-type flowmeter, and acid solution, alkali lye peristaltic pump flow can be regulated through the pump speed-regulating valve, has further improved the sensitivity of pH control, pH control response deviation ± 0.01.
Dissolved oxygen control:
Adopt the standalone module mode, the dissolved oxygen unit is provided with separately.Use the PID master mode to realize control automatically, have Adaptive PID Control adjustment function, the pid parameter that automatically setting is best.Dissolved oxygen control is related with the oxygen ventilation valve, through the valve switch, realizes the automatic control of dissolved oxygen.When dissolved oxygen is lower than set(ting)value, unit start-up control valve, logical oxygen improves dissolved oxygen to set(ting)value in reactor drum.Oxygen flow can be regulated through spinner-type flowmeter, has further improved the sensitivity of dissolved oxygen control, dissolved oxygen response variance 0.5%.
The process pipeline system:
Wherein, Gas system comprises pressurized air gas piping, oxygen gas pipeline, dioxide gas pipeline, gas pressure reducer, and pneumatic diaphragm valve is controlled automatically; Be connected with a jar external circulation line through the gas make-up pipeline respectively, and under the effect of peristaltic pump, import in the tank body.
Liquid system comprises that liquid replenishes pipeline and liquid collecting tube road, is connected with the jar external circulation line respectively, and under the peristaltic pump effect, adds feed liquid in the entering tank body or in tank body, receive liquid.
The process implementing flow process
Cell cultures:
Connect all pipelines by aseptic requirement, inject about cell culture fluid 25L in big jar through peristaltic pump 4, water-bath is preheated to 25-28 ℃ then, and cell and microcarrier directly inject built-in canister through pipeline G, and total liquid is 30L-32L in the at this moment whole jar.Turn off pipeline B and E, low rate start peristaltic pump 4 lets cell fully be attached at (also can outside jar, let cell attach good back injects in the built-in canister) on the microcarrier at circulation some hrs in the built-in canister up to cell; Let temperature slowly be raised to 37 ℃ ± 0.3 ℃ simultaneously, pass through pipeline D behind the circulation some hrs outside the cell drain tank that does not attach, then opening conduits B and E; Make the liquid in big jar also participate in the canister circulation; The process of liquid circulation also is that slowly flowing fluid can not damage by pair cell to the well-beaten process of jar inner cell in the canister, and liquid is mobile from bottom to top; Not only do not have the dead angle, and stir very even.Data according to dissolved oxygen and the detection of pH probe; Can turn off pipeline A, D earlier, circulate to big jar through peristaltic pump injection oxygen and sodium hydrogencarbonate, gas and sodium hydrogencarbonate are not directly followed cells contacting like this; Can avoid the too high pair cell of bubble and local PH to damage; After mixing up, opening conduits A, D circulate, thereby make the interior physical and chemical index of big canister consistent.
The cleaning of cell:
Bleed off pot liquid through pipeline E, D, fill with the washing lotion circulation by peristaltic pump then, bleed off pot liquid through pipeline E, D again after the some time, look repeatedly repeated washing of particular case.
Virus culture:
In big jar, inject about virus-culturing fluid 25L through peristaltic pump, seed culture of viruses directly injects built-in canister through pipeline G, and total liquid is 30L-32L in the at this moment whole jar.Turn off pipeline B and E, the low rate start peristaltic pump lets liquid circulation some hrs in built-in canister make fully cells infected of virus; Let temperature remain on 35 ℃ ± 0.3 ℃, behind the circulation some hrs through pipeline D outside the cell drain tank that comes off, opening conduits B and E then; Make the liquid in big jar also participate in the canister circulation,, can turn off pipeline A, D earlier according to the data of dissolved oxygen and the detection of PH probe; Circulate to big jar through peristaltic pump injection oxygen and sodium hydrogencarbonate; Gas and sodium hydrogencarbonate directly with cells contacting, can not avoid the too high pair cell of bubble and local PH to damage, after mixing up like this; Opening conduits A, D circulate, thereby make the interior physical and chemical index of big canister consistent.After cultivating some hrs, infected viral cell (disposable results) according to different goods results, or collected virus-culturing fluid (keep situation according to cell and can repeatedly repeat to gather in the crops virus-culturing fluid), so just accomplished a production cycle through pipeline E, D.
The present invention can provide the external circulation animal cell culture bioreactor of working volume 30-150L.
The above; Be merely the preferable embodiment of the present invention; But protection scope of the present invention is not limited thereto; Any technician who is familiar with the present technique field is equal to replacement or change according to technical scheme of the present invention and inventive concept thereof in the technical scope that the utility model discloses, all should be encompassed within protection scope of the present invention.
Claims (3)
1. an external circulation animal cell culture bioreactor comprises tank body and external circulation line, well heater, peristaltic pump and system, it is characterized in that:
Said tank body comprises big jar and canister; Said big jar comprises big tank, water-bath interlayer and cover, and said cover is provided with ventage; Said canister comprises lower cover and middle tank body, and the said lower cover of going up is tightly connected with middle tank body respectively; Through being arranged at lower cover and middle tank body junction on the said canister inside and perpendicular to the web plate of stainless steel up and down of the axis of said canister, following three spaces during said canister is divided into;
Said canister is placed on said big jar inside; And fix through pipeline D, A, G and said big jar of cutting ferrule respectively; That is: the interior extreme lower position of lower cover of canister is fixed and be communicated to pipeline D one end; The bottom that its other end is arrogant jar is connected to outside big jar and through threeway and is connected with the I section and the pipeline E of jar external circulation line respectively, and the arrogant tank body of the other end of said pipeline E bottom is inserted in big jar; The extreme higher position that pipeline A one end is fixed in the upper cover of canister also is communicated in the canister since then; The top of the arrogant tank wall of its other end picks out outside the tank body; And be communicated with the II section and the pipeline B of jar external circulation line through threeway; Pipeline B is equipped with diaphragm valve in this port, and the arrogant tank wall of the other end of said pipeline B middle and upper part is connected in big jar; The end of pipeline G is fixed in the upper cover of canister and passes the upper cover of canister and the middle tank body that last web plate is connected to said canister successively, and its other end is switched to outside the big tank;
Said external circulation line comprises pipeline E and the pipeline D that pipeline A that the outer top of a jar external circulation line, tank body is connected with said jar of external circulation line through threeway and pipeline B, the outer bottom of tank body are connected with said jar of external circulation line through threeway; Said jar of external circulation line passes said peristaltic pump; Said peristaltic pump adopts electrodeless variable-speed;
Also connect liquid respectively on the said jar of external circulation line and replenished pipeline, gas make-up pipeline, liquid collecting tube road;
Said pipeline D connects blow-off pipe at its lowest order;
The monitoring probe that has also connected pH control and dissolved oxygen control on the II section of said jar of external circulation line; Be placed with temperature controlled monitoring probe in the said big tank; Above-mentioned monitoring probe is connected with housing through lead, and said housing comprises indicating meter, dissolved oxygen unit, pH unit, temperature regulator, datalogger.
2. external circulation animal cell culture bioreactor according to claim 1 is characterized in that:
The arrogant tank body of said pipeline E bottom is inserted in big jar along big tank wall tangential.
3. external circulation animal cell culture bioreactor according to claim 2 is characterized in that:
The said lower cover of going up is tightly connected through external clip with middle tank body respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010242025 CN101899394B (en) | 2010-07-29 | 2010-07-29 | External circulation animal cell culture bioreactor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010242025 CN101899394B (en) | 2010-07-29 | 2010-07-29 | External circulation animal cell culture bioreactor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101899394A CN101899394A (en) | 2010-12-01 |
| CN101899394B true CN101899394B (en) | 2012-09-26 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010242025 Expired - Fee Related CN101899394B (en) | 2010-07-29 | 2010-07-29 | External circulation animal cell culture bioreactor |
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| CN102229888B (en) * | 2011-06-08 | 2013-06-12 | 南京大学医学院附属鼓楼医院 | Feedback-type pneumatic-control pressure stress cell culture device |
| US8183035B1 (en) * | 2011-09-07 | 2012-05-22 | Therapeutic Proteins International, LLC | Single container manufacturing of biological product |
| CN103589638B (en) * | 2013-11-12 | 2014-12-17 | 罗火生 | Pneumatic self-circulation animal cell culture bioreactor and application method thereof |
| CN110129197A (en) * | 2018-02-08 | 2019-08-16 | 江阴瑞康健生物医学科技有限公司 | A kind of incubator with liquid circulation function |
| CN109652299A (en) * | 2018-11-15 | 2019-04-19 | 上海量能生物科技有限公司 | Bioreactor with resistance to movement function |
| CN110106085B (en) * | 2019-05-24 | 2022-06-28 | 中山大学 | Integrated bioreactor for adherent cell culture |
| CN110551631B (en) * | 2019-10-10 | 2024-03-08 | 南京比瑞生物科技有限公司 | Fixed bed bioreactor system for mass production of mesenchymal stem cells |
| CN111690509A (en) * | 2020-06-11 | 2020-09-22 | 连云港百仑生物反应器科技有限公司 | External circulation cell jar |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101182454A (en) * | 2007-11-05 | 2008-05-21 | 程龙 | External-internal double-cycling bioreactor |
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