CN108424855A - A kind of plant cell, organ liquid culturing apparatus - Google Patents
A kind of plant cell, organ liquid culturing apparatus Download PDFInfo
- Publication number
- CN108424855A CN108424855A CN201810398376.0A CN201810398376A CN108424855A CN 108424855 A CN108424855 A CN 108424855A CN 201810398376 A CN201810398376 A CN 201810398376A CN 108424855 A CN108424855 A CN 108424855A
- Authority
- CN
- China
- Prior art keywords
- culture
- tank body
- air inlet
- inoculation mouth
- mouth
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
-
- 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
- C12M23/22—Transparent or translucent parts
-
- 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
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- 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
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
-
- 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
- C12M29/20—Degassing; Venting; Bubble traps
-
- 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
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/02—Filters
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention provides a kind of easy plant cell and organ liquid culturing apparatus, including hollow culture tank body, are located at the inoculation mouth and air inlet of culture tank body one end, and the exhaust outlet positioned at the tank body other end；Wherein, described tank body one end is plane, and one end is taper, and inoculation mouth, inoculation mouth fitted seal lid is arranged in the top of taper.Compared with existing culture apparatus, culture tank volume of the invention is 2 100L, easy to operate, operates repeatability by force, of low cost, and dangerous small, contamination rate is low；The materials'use of culture tank body and key fitment can directly be sterilized with heat safe transparent material, and culture medium capacity is stablized, and incubation is intuitively visible；Air inlet is equipped with air sparger and is obliquely installed, and carries out rotary air inlet, and the needs of culture medium dissolved oxygen can be just met under low pass tolerance, reduces the loss of gas supply cost and culture medium；And such as adventitious root of the organ for being difficult to collect, equipment are furnished with special collection port.
Description
Technical field
The present invention relates to technical field of cell culture, and in particular to a kind of a kind of plant cell of suitable large-scale production,
Organ liquid culturing apparatus.
Background technology
With world rapid economic development, the ample supply of the fossil resources such as existing oil coal becomes to hold
It is continuous, it is difficult to support the advanced developing goal of human society.Human social development will be from dependent on hydrocarbons such as fossil fuels
Resource is changed into the carbon aquation that solar continuous synthesis can be utilized by starch, cellulose, polysaccharide, plant and microbial grease etc.
Close goods and materials source.The transformation of this energy and resource structures will be that biotechnology and its industry development bring very big opportunities and challenges.
For plant available resources, China is one of the country that resources of medicinal plant is most abundant in the world, with plant
A large amount of uses of medicine, the excavation of medicinal plant and using the speed of growth considerably beyond plant, resources of medicinal plant is increasingly withered
It exhausts.And traditional medicinal plant field production is influenced by " favourable climate and geographical position ", and output and quality fluctuation is big, and pesticide residual contamination is asked
Topic is also than more serious.
Therefore, people are by means of various biosystems, using carbohydrate come all needed for large-scale production modern society
Such as plant cell, tissue and the various useful components of organ.And the biochemical reaction that biosystem carries out substance conversion is in biology
It is carried out in the microhabitat of this relative closure of reactor.Bioreactor provides controllable for the biochemical reaction of biosystem
Environmental condition promotes biomass conversion process efficiently to carry out, for example, temperature, pH, dissolved oxygen, mixing, shearing, feed supplement etc..It passes
System bioreactor generally comprises suspension culture systems reactor and Immobilized culture system response device.The former includes mainly stirring
Formula reactor, airlift reactor；The latter includes mainly membrane reactor, packed bed reactor.
The bioreactor of culture plant cell, tissue and organ has arrived the stage of industrialized production at present.Example
Such as, with airlift bioreactor culture Catharanthus Roseus Cell production serfin [A.H.Scragg et al., Enzyme Microb
Technol, 1987,9：619]；South Korea S.H.Son et al. to yew cell carry out reactor large-scale culture, using from
Airlift reactor [S.H.Son et al., the Plant Cell Reports (2000) 19 of 100L to 500L：628-633]；
After several years, Kee-Yoeup Paek carry out large-scale culture to ginseng adventitious root using spherical bubbling style reactor and obtain with very
Ginsenoside [Kee-Yoeup Paek etal., Adv the Biochem Engin/Biotechnol (2009) of strong pharmacological activity
113：151-176], obtain the ginseng adventitious root etc. compared with height increament.Meiyan Inst. of Biological Engineering, Meixian County is followed using gas
Ring type bioreactor production yew cell has been also up to level [the gas circulating type plant cell reactor of 500L
CN2510499Y].U.S. Phyton, Inc have been reached using stirring type bioreactor culture yew cell production taxol
To 7 tons of scale.South Korea SamyangGenex is also producing taxol using stirred-tank fermenter culture yew cell.
Stirring reactor is one of most commonly used reactor of industrial applications, due to plant cell have volume compared with
Greatly, cell wall is more crisp and has the characteristics that big vacuole, and plant cell is very sensitive to shearing force, sensibility by cell kind
The many factors such as class, state, shearforce tolerance, the structure of shear rate and reactor itself determine.Currently, the approach solved
Typically from cell line, the high yield system of anti-shearing force is screened or by cell engineering and genetic engineering means to existing
Cell line is transformed, and establishes the strain of anti-shearing force；Structure of reactor is transformed on this basis, makes it have the flow field of mitigation
And good mixed performance.But still it has the following problems：1) currently used reactor is all integrally fixed at a certain position, inoculation
When need outdoor inoculation, while secondary opening tank body is needed when feed supplement, microbiological contamination probability can obviously increase, and microbiological contamination problem is plant
It is most difficult to solve the problems, such as in tissue cultures；2) tank body that reactor is mostly made using stainless steel as material, therefore all needed when making
It is arranged one and arrives multiple transparent viewing ports.Many dead angles are still had in actual use to be difficult to observe by；3) stirring-type biology
Reactor opereating specification is big, realizes the also more of industrialization production, but the shearing force generated in whipping process is larger, is easy to planting
Object in vitro tissue causes to damage, and energy consumption is larger；4) since mechanical stirring device cannot sterilize, the type equipment is gone out
It needs steam-out (SO) to sterilize when bacterium, pays particular attention to the danger and complexity of high steam, and steam sterilizing goes out
Bacterium can cause culture volume to increase simultaneously, and it is unstable to accelerate, also result in substance volatile in culture medium and
Gasification temperature is discharged less than the substance of water with steam；
In contrast, airlift reactor and bubbling style reactor shearing force are smaller, generally believe bubbling both at home and abroad at present
Formula reactor is a kind of widely applicable bioreactor, and this kind of reactor is structurally characterized in that bubble rises along liquid phase, with
Agitation liquid is to improve mass transfer rate while liquid phase contact is reacted.However the advantages of being in theory above, really
Realize that the very few of industrialization, most of bubbling style reactors only remove the mixing plant of stirring reactor, because
This its use scope is far below stirred-tank fermenter, and traditional bubbling style reactor is returned due to being susceptible to without agitating device
It is mixed serious, phenomena such as bubble is also easy to produce coalescence, and foam can carry some 2 volatile materials of CO secretly, while in order to which culture is molten
The demand of oxygen needs largely to be passed through gas, and intercourse meeting leads to a large amount of evaporations of culture medium, is generated not to the growth of plant cell
Good influence, and it is less efficient, although can weaken this influence by the way that antifoaming agent is added, antifoaming agent is added can be thin to plant
Born of the same parents, the growth of tissue and organ and the accumulation of secondary metabolite generate bad.Simultaneously because air inlet is to dissolved oxygen in culture medium
Incrementss are insufficient, therefore culture medium dissolved oxygen problem is than more serious, while the excessive material for causing some to need long-term cultivation of ventilatory capacity
Material, later stage can have to terminate culture because of the evaporation of culture medium.Many equipment of the type and stirring type bioreactor one
Sample needs to sterilize using external high steam, increases danger and complexity when operation, and culture medium holds when sterilizing
The unstability of amount.
The reactor of any of the above type is based on amplification scale, due to needing to amplify step by step, centre any one
Go wrong such as microbiological contamination, browning of step can lead to the failure finally cultivated, and mini-reactor simple to operation can be with
Problem above is solved well.
In small-sized biological reactor simple to operation, that has been applied at present has GE Healthcare Wave
The wave bioreactor of Products Group productions, WAVE wave bioreactor cultures volume range is flexible, operates
Simply, control precision is reliable, easy to technique amplification.But wave wave reactors use composite material, are used as fermentation tank
Polybag is combined by three-layer plastic, internal layer LDPE, and centre is EVOH, and outermost layer is EVA materials, cannot be gone out into horizontal high voltage
Bacterium needs that culture bag is first needed irradiation sterilization when in use, and then culture medium pours into after individually sterilizing in culture bag, and centre is such as
Fruit needs feed supplement to need to reopen culture bag and operated, and adventitious root class culture is difficult to cultivate in wave reactors,
And culture bag can not reuse, therefore environmental pollution can be caused, in addition, the oxygen of wave culture bag culture mediums is to pass through
Liquid surface gas exchanges carry out, therefore dissolved oxygen is easy deficiency in culture medium, and culture late growth will be severely limited, most
Culture quality is caused to go wrong with yield eventually.
Complicated for operation to solve equipment costliness in the prior art, culture medium evaporation capacity is big in incubation, incubation
In largely generate bubble, feed supplement equipment, culture medium and culture device separately sterilize, the problem of increasing microbiological contamination when feed supplement, the present invention
Dissolved oxygen demand can be met under a kind of quasi- low pass gas of offer, feed supplement equipment can simultaneously sterilize together with culture device, incubation
The lower incubator of middle bubble formation rate.
Invention content
The object of the present invention is to provide a kind of plant cells and organ liquid culturing apparatus.
In order to solve the above technical problems, the present invention adopts the following technical scheme that：
A kind of plant cell and organ liquid culturing apparatus, including hollow culture tank body are located at culture tank body one end
Inoculation mouth and air inlet, and the exhaust outlet positioned at the tank body other end；Wherein, described tank body one end is plane, and one end is taper,
Inoculation mouth, inoculation mouth fitted seal lid is arranged in the top of taper.
The present invention biotic component liquid culturing apparatus using can high-temperature sterilization material make, be especially resistant to 121
DEG C autoclaved material, more preferable transparent material, such as PC, PE, glass etc. in this way can directly put accessory and culture tank body
Enter in autoclaving equipment and sterilizes.Its application method is as follows：In inoculation, forward direction is placed, and plane is located at lower section, closing row
Gas port is introduced directly into from inoculation mouth in culture medium after shaking up a certain amount of cell together with culture solution；In culture, inoculation mouth is close
Envelope, tank body is inverted, air inlet is made to be located at lower section, exhaust outlet is located above, and is passed through gas, makes gas using the uphill process of bubble
Bubble comes into full contact with the culture solution in reacting tank body.
Consequently, to facilitate placing, the exhaust outlet is positioned close to the side of its bottom end plane.
Further, the air inlet may be provided on the inclined-plane of taper tank body, and the air inlet connects air inlet pipe, will train
Required gas is supported to import in culture tank body.
Further, the air inlet is arranged on sealing cover, can effectively reduce leakage in this way, reduces sealing difficulty.
Further, air inlet is arranged using inclined mode, and the gas out of plumb being discharged into is made to increase；
Further, the air inlet is additionally provided with air sparger, to promote gas presentation to circulate state
It is non-perpendicular upward, extend running orbit of the bubble in cultivating tank body.The air sparger can be such that bubble diameter greatly reduces,
It is preferred that air sparger of the Air Bubble Size between 0.2mm-8mm can be made.
Further, gas filtration sterilization device is set at the gas access of the air inlet pipe, it is preferably commonly used in the art
The air filtration sterilization device of 0.22um, prevents the miscellaneous bacteria in air from entering culture tank.
Further, the inoculation mouth is spiral mouth, and the sealing cover is twoport helixseal lid, connection air inlet suitable for reading
Pipe, the inoculation mouth of lower mouth connection culture tank body.
The inoculation mouth actual aperture size is depending on culture state, it is contemplated that be mainly used for cultivate plant cell and
Organ, inoculation mouth diameter can be 2-15cm.
Further, between the angle of the circular cone of the culture tank body is 60-150 DEG C.
Further, the inoculation mouth is cylinder, and height is no more than 20cm.
In seeded process, cultivates culture medium height in bucket and be greater than 20cm；In incubation, culture medium liquid level and row
It is greater than 3cm between gas port.
The exhaust outlet can be used for feed supplement in culture midway；Or exhaust outlet separates branch pipe(tube) for feed supplement；
Further, the side close to exhaust outlet of the culture tank body is additionally provided with one or more opening,
The opening is used for feed supplement, connects one or more feed supplement pump.
After culture, culture can be poured out from inoculation mouth；
Or it is further, collection port is arranged in culture tank body side surface.Such as encounter that the culture for being difficult to collect is for example agglomerating or ball is difficult
With the culture poured out from inoculation mouth, or large-scale culture such as adventitious root, can be collected using collection port, in actual use
Determine whether to be equipped with according to culture state.The collection port is being inoculated with and is being sealed in incubation.
In addition, to prevent living contaminants, the outlet of the exhaust outlet to be provided with filtration sterilization component, such as simple gauze or
Cotton etc. prevents the microorganism of outside from entering culture device from exhaust outlet.
It cultivates tank body when sterilizing just to put, the devices such as exhaust ports tongs of bottom are closed, and inoculation mouth uses sterile envelope
Membrana oralis is sealed, the sealing cover and air inlet pipe of inoculation mouth, and air sparger and air disinfector connect together, then
It sterilizes.
When in use, after culture seed being poured into tank body together with partial medium, by inoculation mouth seal cap sealing,
It sets and is placed on culturing rack with holes, for culture device on culturing rack, making inoculation mouth downward, exhaust outlet is upward, connects upper air
The tongs that pipe is opened on exhaust pipe can ventilate, according to oneself needing to adjust ventilatory capacity size when use.
The capacity of the culture tank body of the present invention is 2-100L, is highly convenient for operating, sterilizing, which can also be directly placed into, commonly goes out
Bacterium pot sterilizes.
The object of the present invention is to provide a kind of easy plant cell and organ liquid culturing apparatus, main advantage are as follows：
1) it cultivates tank body and the materials'use of key fitment can be with heat safe transparent material, such as PC, PE, glass etc., it can
It directly sterilizes, when sterilizing is positioned in common autoclave, need not be passed through steam, therefore culture medium capacity is stable；Training
The process of supporting is intuitive as it can be seen that culture can be observed obviously in any corner of cylinder in incubation, cultivates the growth of bucket
State is very clear, when there is phenomena such as microbiological contamination, both can early find to handle as early as possible before sporogenesis, reduces to it
The pollution of his cylinder；
2) air inlet is equipped with air sparger, and air inlet is discharged into gas using inclined mode, carries out rotary air inlet, gas
While bubble drives liquid vertical ascent, liquid can also be driven to be rotated along bucket wall, promote gas between culture medium and bubble
It exchanges, realizes and meet the needs of culture medium dissolved oxygen under low pass tolerance, therefore can not be seldom or even in practical incubation
Use antifoaming agent；And then reduce in venting process and injured caused by culture in the bubble for cultivating the formation of its surface, reduce gas supply
Cost；The loss of culture medium caused by ventilation evaporation can also substantially reduce, and the proportion of goods damageds of culture medium can be down to 10% or less；
3) tank body and feed supplement equipment can sterilize simultaneously, reduce the secondary operation that tank body is opened when feed supplement, and then reduce dirty
Dye rate；
4) for the culture for the organ such as adventitious root for being difficult to collect, equipment is furnished with special collection port, greatly reduces culture
Collection work；
5) volume selects 2-100L capacity, easy to operate, and when inoculation, which is positioned on aseptic operating platform, to carry out, and greatly reduces and connects
Contamination rate when kind；Operating process is simple and convenient, and dangerous small, contamination rate is low, operates repeatability by force, of low cost, can be significantly
Reduce the production cost of culture.
Description of the drawings
Fig. 1 is the schematic diagram of the plant cell and organ liquid culturing apparatus described in the embodiment of the present invention 1；
Fig. 2 is the schematic diagram of the plant cell and organ liquid culturing apparatus described in the embodiment of the present invention 2；
Wherein, 1, culture tank body；2, inoculation mouth；3, sealing cover；4, air inlet；5, exhaust outlet；6, material-feeding port；7, air point
Cloth device；8, air filter；9, cotton is filtered；10, collection port.
Specific implementation mode
Plant cell of the present invention and organ liquid culturing apparatus are further described below in conjunction with the accompanying drawings.
Embodiment 1
A kind of plant cell and organ liquid culturing apparatus, including culture tank body 1 are (autoclaved by being resistant to 121 DEG C
PC is made, capacity 40L), it is located at the inoculation mouth 2 and air inlet 4 of 1 one end of culture tank body, 2 a diameter of 10cm of inoculation mouth；And
Positioned at the exhaust outlet of the tank body other end；Wherein, the tank body is cylinder taper, and the angle of the circular cone of the culture tank body 1 is 64
DEG C, inoculation mouth 2 is arranged in the top of taper, and the height of inoculation mouth 2 is 12cm, 2 fitted seal lid 3 of inoculation mouth；The exhaust outlet is set
Set the side close to its bottom surface in cylinder；The air inlet 4 is arranged on sealing cover 3, and the inoculation mouth 2 is spiral mouth, institute
It is twoport helixseal lid, connection air inlet pipe suitable for reading, the inoculation mouth 2 of lower mouth connection culture tank body 1 to state sealing cover 3.
Air inlet 4 is arranged using inclined mode, and the gas out of plumb being discharged into is made to increase；The air inlet 4 is additionally provided with
Air sparger 7 extends fortune of the bubble in cultivating tank body 1 to promote gas presentation to circulate state rather than vertically upward
Row track.The air sparger 7 makes bubble diameter in 0.2mm-8mm.
The air filter 8 of 0.22um is set at the gas access of the air inlet pipe, and the outlet of the exhaust outlet 5 is provided with
Filtering cotton 9 is for preventing miscellaneous bacteria from entering culture tank body 1.In addition, the exhaust outlet 5 separates branch pipe(tube) as material-feeding port 6, it is used for
Feed supplement；
Its application method is as follows：Tank body 1 to be cultivated when sterilizing just to put, the devices such as exhaust ports tongs of bottom are closed,
Inoculation mouth 2 is sealed using sterile sealed membrane, and the sealing cover 3 and air inlet pipe of inoculation mouth 2, air sparger 7 and air remove
Bacterium device, connects together, and then sterilizes；After sterilizing, culture tank body 1 is positive to be placed, and cylinder is located at lower section, closes
Exhaust outlet injects culture medium in culture tank body 1, direct from inoculation mouth 2 after then shaking up a certain amount of cell together with culture solution
It imports in culture medium；Then inoculation mouth 2 seals, and inversion is placed on culturing rack with holes, and culture device is on culturing rack, making to connect
Kind mouth 2 is downward, and exhaust outlet 5 is upward, and the tongs that connection intake pipe is opened on exhaust pipe can ventilate, and utilize bubble
Uphill process makes bubble be come into full contact with the culture solution in reacting tank body, according to oneself needing to adjust ventilatory capacity size when use.
In seeded process, cultivates culture medium height in bucket and be greater than 20cm；In incubation, between culture medium liquid level and exhaust outlet
It is greater than 3cm.
After culture, culture is poured out from inoculation mouth 2.
Embodiment 2
A kind of plant cell and organ liquid culturing apparatus, including culture tank body 1 are ((autoclaved by being resistant to 121 DEG C
PC is made, capacity 100L), it is located at the inoculation mouth 2 and air inlet 4 of 1 one end of culture tank body, 2 a diameter of 10cm of inoculation mouth；And
Positioned at the exhaust outlet of the tank body other end；Wherein, the tank body is cylinder taper, and the angle of the circular cone of the culture tank body 1 is 120
DEG C, inoculation mouth 2 is arranged in the top of taper, and the height of inoculation mouth 2 is 8cm, 2 fitted seal lid 3 of inoculation mouth；The exhaust outlet setting
In cylinder close to the side of its bottom surface；The air inlet 4 is arranged on the inclined-plane of taper tank body, the air inlet 4 connect into
Tracheae imports the gas needed for culture in culture tank body 1.
The side of cylinder close to the exhaust outlet 2 of the culture tank body 1 is also set up there are one material-feeding port 6, for connecting one
Or more than one feed supplement pump, the feed supplement in incubation.
The side of culture tank body 1 is additionally provided with the collection port 10 of a diameter of 30cm.Such as encounter the culture for being difficult to collect such as
Agglomerating or ball is difficult to the culture poured out from inoculation mouth 2, or large-scale culture such as adventitious root, can be received using collection port 10
Collection.The collection port 10 is being inoculated with and is being sealed in incubation.
Air inlet 4 is arranged using inclined mode, and the gas out of plumb being discharged into is made to increase；The air inlet 4 is additionally provided with
Air sparger 7 extends fortune of the bubble in cultivating tank body 1 to promote gas presentation to circulate state rather than vertically upward
Row track.The air sparger 7 makes bubble diameter in 0.2mm-8mm.
The air filter 8 of 0.22um is set at the gas access of the air inlet pipe, and the outlet of the exhaust outlet 5 is provided with
Filtering cotton 9 is for preventing miscellaneous bacteria from entering culture tank body 1.
It is identical as process described in embodiment 1 using process.
The culture tank volume of the present invention is 2-100L, easy to operate, operates repeatability by force, of low cost, dangerous small, dye
Bacterium rate is low；The materials'use of culture tank body and key fitment can directly be sterilized with heat safe transparent material, culture medium capacity
Stablize, incubation is intuitively visible；Air inlet is equipped with air sparger and is obliquely installed, and carries out rotary air inlet, can be low
Just meet the needs of culture medium dissolved oxygen under ventilatory capacity, reduces the loss of gas supply cost and culture medium；And for being difficult to collect
Organ such as adventitious root, equipment are furnished with special collection port.
Claims (10)
1. a kind of plant cell and organ liquid culturing apparatus, which is characterized in that including hollow culture tank body, be located at culture tank
The inoculation mouth and air inlet of body one end, and the exhaust outlet positioned at the tank body other end；Wherein, described tank body one end be plane, one
End is taper, and inoculation mouth, inoculation mouth fitted seal lid is arranged in the top of taper.
2. culture apparatus as described in claim 1, which is characterized in that the culture apparatus is gone out using can tolerate 121 DEG C of high pressures
The material of bacterium is made；The 121 DEG C of autoclaved materials that can tolerate of preferably clear are made.
3. culture apparatus as claimed in claim 1 or 2, which is characterized in that the exhaust outlet is positioned close to its flat bottom
Side；The air inlet is arranged on the inclined-plane of taper tank body or is arranged on sealing cover, and the air inlet connects air inlet pipe,
Gas needed for culture is imported in culture tank body.
4. culture apparatus as claimed in claim 1 or 2, which is characterized in that a diameter of 2-15cm of inoculation mouth.
5. culture apparatus as claimed in claim 3, which is characterized in that the air inlet is arranged using inclined mode, described
Air inlet is provided with air sparger.
6. culture apparatus as claimed in claim 3, which is characterized in that the air inlet is arranged on sealing cover, the inoculation
Mouth is spiral mouth, and the sealing cover is twoport helixseal lid, connection air inlet pipe suitable for reading, the inoculation of lower mouth connection culture tank body
Mouthful.
7. culture apparatus as described in claim 1, which is characterized in that the angle of the taper of the culture tank body is 60-150
℃。
8. culture apparatus as described in claim 1, which is characterized in that the side close to exhaust outlet of the culture tank body is also set
It is equipped with one or more opening, the opening is used for feed supplement, connects one or more feed supplement pump.
9. culture apparatus as described in claim 1, which is characterized in that collection port is arranged in the tank body side of the culture tank body.
10. culture apparatus as described in claim 1, which is characterized in that gas mistake is arranged at the gas access of the air inlet pipe
Sterilization device is filtered, the outlet of the exhaust outlet is provided with filtration sterilization component；The capacity of the culture tank body is 2-100L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810398376.0A CN108424855A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ liquid culturing apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810398376.0A CN108424855A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ liquid culturing apparatus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108424855A true CN108424855A (en) | 2018-08-21 |
Family
ID=63162070
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810398376.0A Pending CN108424855A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ liquid culturing apparatus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108424855A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108641950A (en) * | 2018-04-28 | 2018-10-12 | 大连普瑞康生物技术有限公司 | A kind of bioreactor of culture plant cell |
| CN117256478A (en) * | 2023-11-22 | 2023-12-22 | 上海科缔联科技有限公司 | Continuous fermentation culture system for plant tissues |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2301847Y (en) * | 1997-07-14 | 1998-12-30 | 中国科学院化工冶金研究所 | Plant organ biological culture reactor |
| US20050032211A1 (en) * | 1996-09-26 | 2005-02-10 | Metabogal Ltd. | Cell/tissue culturing device, system and method |
| CN102321537A (en) * | 2011-08-31 | 2012-01-18 | 天津大学 | Bubbling bioreactor used for plant tissue culture |
| CN102517199A (en) * | 2011-11-17 | 2012-06-27 | 西安建筑科技大学 | Multi-cycle stirring egg-shaped anaerobic fermentation system |
-
2018
- 2018-04-28 CN CN201810398376.0A patent/CN108424855A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050032211A1 (en) * | 1996-09-26 | 2005-02-10 | Metabogal Ltd. | Cell/tissue culturing device, system and method |
| CN2301847Y (en) * | 1997-07-14 | 1998-12-30 | 中国科学院化工冶金研究所 | Plant organ biological culture reactor |
| CN102321537A (en) * | 2011-08-31 | 2012-01-18 | 天津大学 | Bubbling bioreactor used for plant tissue culture |
| CN102517199A (en) * | 2011-11-17 | 2012-06-27 | 西安建筑科技大学 | Multi-cycle stirring egg-shaped anaerobic fermentation system |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108641950A (en) * | 2018-04-28 | 2018-10-12 | 大连普瑞康生物技术有限公司 | A kind of bioreactor of culture plant cell |
| CN108641950B (en) * | 2018-04-28 | 2024-01-02 | 大连普瑞康生物技术有限公司 | Bioreactor for culturing plant cell line |
| CN117256478A (en) * | 2023-11-22 | 2023-12-22 | 上海科缔联科技有限公司 | Continuous fermentation culture system for plant tissues |
| CN117256478B (en) * | 2023-11-22 | 2024-02-02 | 上海科缔联科技有限公司 | Continuous fermentation culture system for plant tissues |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN203048953U (en) | Dismountable plant tissue fluid culture device | |
| CN106520552B (en) | A kind of biological reactor for cell culture | |
| CN201459137U (en) | Microbial dialysis incubator | |
| CN102321537B (en) | Bubbling bioreactor used for plant tissue culture | |
| CN104770304B (en) | Plant bioreactor and its application method | |
| CN208857315U (en) | A kind of plant cell and organ Liquid Culture bag | |
| CN108424855A (en) | A kind of plant cell, organ liquid culturing apparatus | |
| CN208857314U (en) | A kind of plant cell, organ liquid culturing apparatus | |
| CN103224882A (en) | Disposable wave type bioreactor with aerating apparatus | |
| CN110437973A (en) | A kind of culture apparatus and cultural method of anaerobic bacteria | |
| CN102816689B (en) | Device and method for flat membrane oxygen-supply self-circulation-type microbial culture | |
| CN102234205B (en) | Multifunctional solid state fermentation reactor | |
| CN101988075B (en) | Method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum | |
| CN108441425A (en) | A kind of plant cell, organ culture device | |
| CN203057904U (en) | Simple fermentation device for liquid strain submerged cultivation | |
| CN207632812U (en) | A kind of acidophilus Propiram fermentation of bacillus system | |
| CN201762330U (en) | Animal cell culture bioreactor | |
| CN202415569U (en) | Tidal animal cell culture bioreactor | |
| Kalaichelvan et al. | Bioprocess technology | |
| CN103125269A (en) | Simple fermentation device for liquid spawn submerged cultivation | |
| CN209481660U (en) | A kind of microalgae carbon sequestration is at mine coproduction integrated apparatus | |
| CN109251847A (en) | Utilize the device and method of sunlight culture photosynthetic microorganism | |
| CN202968544U (en) | Continuous fermentation propagation device for reducing contamination probability | |
| CN206545025U (en) | Liquid strain culturing device and liquid bacterial strain production system | |
| CN1473924A (en) | Ferment method of solid ferment adseptic manipulation and its special device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 116000 Changdianbao Community, Pulandian Economic Development Zone, Dalian, Liaoning Province Applicant after: Puruikang Biotechnology Co., Ltd., Dalian Address before: 116023 No. 32 Lixian Street, Qixianling, Dalian High-tech Industrial Park, Liaoning Province Applicant before: Puruikang Biotechnology Co., Ltd., Dalian |
|
| CB02 | Change of applicant information |