CN108441425A - A kind of plant cell, organ culture device - Google Patents
A kind of plant cell, organ culture device Download PDFInfo
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- CN108441425A CN108441425A CN201810398091.7A CN201810398091A CN108441425A CN 108441425 A CN108441425 A CN 108441425A CN 201810398091 A CN201810398091 A CN 201810398091A CN 108441425 A CN108441425 A CN 108441425A
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- 210000000056 organ Anatomy 0.000 title claims abstract description 15
- 238000011081 inoculation Methods 0.000 claims abstract description 40
- 230000001954 sterilising effect Effects 0.000 claims abstract description 17
- 238000009630 liquid culture Methods 0.000 claims abstract description 9
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 7
- 239000012780 transparent material Substances 0.000 claims abstract description 5
- 238000007789 sealing Methods 0.000 claims description 24
- 239000006052 feed supplement Substances 0.000 claims description 13
- 238000001914 filtration Methods 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 230000008676 import Effects 0.000 claims description 2
- 239000007789 gas Substances 0.000 abstract description 43
- 239000001963 growth medium Substances 0.000 abstract description 24
- 238000011534 incubation Methods 0.000 abstract description 10
- 239000000463 material Substances 0.000 abstract description 10
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 8
- 239000001301 oxygen Substances 0.000 abstract description 8
- 229910052760 oxygen Inorganic materials 0.000 abstract description 8
- 238000011109 contamination Methods 0.000 abstract description 4
- 210000004027 cell Anatomy 0.000 description 27
- 241000196324 Embryophyta Species 0.000 description 26
- 238000000034 method Methods 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000010008 shearing Methods 0.000 description 6
- 230000005587 bubbling Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000011169 microbiological contamination Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 230000008020 evaporation Effects 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 229920000742 Cotton Polymers 0.000 description 3
- 239000002518 antifoaming agent Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000003519 ventilatory effect Effects 0.000 description 3
- 241000208340 Araliaceae Species 0.000 description 2
- 229930012538 Paclitaxel Natural products 0.000 description 2
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 2
- 235000003140 Panax quinquefolius Nutrition 0.000 description 2
- 238000005842 biochemical reaction Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 235000008434 ginseng Nutrition 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229960001592 paclitaxel Drugs 0.000 description 2
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 240000001829 Catharanthus roseus Species 0.000 description 1
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Diphosphoinositol tetrakisphosphate Chemical compound OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000004581 coalescence Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 238000002309 gasification Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229940089161 ginsenoside Drugs 0.000 description 1
- 229930182494 ginsenoside Natural products 0.000 description 1
- WHWDWIHXSPCOKZ-UHFFFAOYSA-N hexahydrofarnesyl acetone Natural products CC(C)CCCC(C)CCCC(C)CCCC(C)=O WHWDWIHXSPCOKZ-UHFFFAOYSA-N 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000010907 mechanical stirring Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000011819 refractory material Substances 0.000 description 1
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000013022 venting Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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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
- 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
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
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- 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
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- 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
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Abstract
The present invention provides a kind of easy plant cell and organ Liquid Culture bag, including culture bag and the handle in culture bag bottom surface one end is set, is arranged in the gas outlet of culture bag bottom end, and inoculation mouth and air inlet in the culture bag other end are set；Wherein, one end of the culture bag setting inoculation mouth is wedge shape, inoculation mouth fitted seal lid.Compared with existing culture bag, culture bag of the invention product is 2 100L, easy to operate, operates repeatability by force, of low cost, and dangerous small, contamination rate is low；The materials'use transparent material of culture bag and key fitment, can directly sterilize or irradiation sterilization, culture medium capacity are 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.
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 Culture bag.
Background technology
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 culture bag is needed when feed supplement, microbiological contamination probability can obviously increase, and microbiological contamination problem is to plant
It is most difficult to solve the problems, such as in object tissue cultures；2) culture bag that reactor is mostly made using stainless steel as material, therefore when making
It is required for setting one to arrive multiple transparent viewing ports.Many dead angles are still had in actual use to be difficult to observe by；3) stirring-type
Bioreactor operation range is big, realizes the also more of industrialization production, but the shearing force generated in whipping process is larger, is easy
Vitro Plant tissue is caused to damage, and energy consumption is larger；4) since mechanical stirring device cannot sterilize, the type is set
It needs steam-out (SO) to sterilize when standby sterilizing, pays particular attention to the danger and complexity of high steam, and steam goes out
Bacterium sterilizing can cause culture volume to increase simultaneously, and it is unstable to accelerate, and also results 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.
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 Culture bag.
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 Culture bag, including culture bag and the handle in culture bag bottom surface one end is set,
And inoculation mouth and air inlet in the culture bag other end are set, and be arranged in the gas outlet of culture bag bottom end；Wherein, described
One end that inoculation mouth is arranged in culture bag is wedge shape, inoculation mouth fitted seal lid.
The culture bag of the present invention is made of transparent material, can observe growth shape of the culture in incubation in this way
State, if 121 DEG C of high pressure sterilizations can be carried out using heat-resisting material, if spoke can be carried out using the non-refractories such as PE material
It can integrally sterilize in sterilizing according to sterilizing.
Application method is as follows：Inoculation when, forward direction place, inoculation mouth upward, close gas outlet, by a certain amount of cell together with
Culture solution is introduced directly into from inoculation mouth in culture medium after shaking up；In culture, inoculation mouth seal cap sealing will be cultivated with handle
Bag is hung up properly, and culture bag is inverted, and air inlet is located at lower section, and gas outlet is located above, and then passes to gas, utilizes the rising of bubble
Journey makes bubble be come into full contact with the culture solution reacted in culture bag.
Therefore, for convenience of using, the gas outlet is positioned close to the side of bottom surface；The air inlet setting is trained in wedge shape
It supports on the inclined-plane of bag or is arranged on sealing cover, the air inlet connects air inlet pipe, and the gas needed for culture is imported culture bag
In..
Further, the air inlet may be provided on the inclined-plane of wedge-shaped culture bag, and the air inlet connects air inlet pipe, will
Gas needed for culture imports in culture bag.
Alternatively, the air inlet is arranged on sealing cover, leakage can be effectively reduced in this way, reduce 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 culture bag.The air sparger can be such that bubble diameter greatly reduces, excellent
Choosing can make air sparger of the Air Bubble Size between 0.2mm-8mm.
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 helixseal lid, and sealing cover is internally provided with silicon
Cushion rubber, to ensure the sealing of sealing cover.
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 2-20cm.
Further, between the angle of the wedge shape of the culture bag is 60-150 DEG C.
Further, the inoculation mouth is generally cylindrical, 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 with go out
It is greater than 3cm between gas port.
The gas outlet can be used for feed supplement in culture midway；Or gas outlet separates branch pipe(tube) for feed supplement；
Further, the culture bag gas outlet connects one or more opening, or close to gas outlet
Side is additionally provided with one or more opening, and the opening is used for feed supplement, connects one or more feed supplement pump.
The opening diameter is 0.2-4cm.
After culture, culture can be poured out from inoculation mouth；
In addition, to prevent living contaminants, the outlet of the gas 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 gas outlet.
Culture bag is just put when sterilizing, is closed with devices such as tongs at the gas outlet of bottom, and inoculation mouth uses sterile sealing
Film is sealed, the sealing cover and air inlet pipe of inoculation mouth, and air sparger and air disinfector connect together, then into
Row sterilizing.
When in use, after culture seed being poured into culture bag together with partial medium, by inoculation mouth seal cap sealing,
Inversion is placed on culturing rack with holes, and for culture device on culturing rack, making inoculation mouth downward, gas outlet is upward, and connection is enterprising
The tongs that tracheae is opened on exhaust pipe can ventilate, according to oneself needing to adjust ventilatory capacity size when use.
The capacity of the culture bag of the present invention is 2-100L, and culture bag can fold, be highly convenient for operating.
The object of the present invention is to provide a kind of easy plant cell and organ Liquid Culture bag, main advantage is as follows：
1) the materials'use transparent material of culture bag and key fitment, such as PC, PE etc. directly can integrally sterilize；It cultivated
Cheng Zhiguan cultivates the growth conditions of bucket as it can be seen that culture can be observed obviously in any corner of cylinder in incubation
It is very clear, it when there is phenomena such as microbiological contamination, both can early find to handle as early as possible before sporogenesis, reduce to other cylinders
The pollution of body；
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) culture bag and feed supplement equipment can sterilize simultaneously, open the secondary operation of culture bag when reducing feed supplement, and then subtract
Of low pollution rate；
4) 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 Culture bag described in the embodiment of the present invention 1；
Wherein, 1, culture bag；2, inoculation mouth；3, sealing cover；4, air inlet；5, gas outlet；6, material-feeding port；7, air is distributed
Device；8, air filter；9, cotton is filtered；10, handle.
Specific implementation mode
Plant cell of the present invention and organ Liquid Culture bag are further described below in conjunction with the accompanying drawings.
Embodiment 1
A kind of plant cell and organ Liquid Culture bag, including culture bag 1 (by can PE be made, capacity 40L), including training
It supports bag 1 and the handle 10 in 1 bottom surface one end of culture bag is set, and inoculation mouth 2 and air inlet in 1 other end of culture bag are set
4,2 a diameter of 10cm of inoculation mouth；It is arranged in the gas outlet 5 of 1 bottom end of culture bag；Wherein, inoculation mouth 2 is arranged in the culture bag 1
One end be wedge shape, the culture bag 1 wedge shape angle be 64 DEG C, wedge-shaped top be arranged inoculation mouth 2, the height of inoculation mouth 2
Degree is 12cm, 2 fitted seal lid 3 of inoculation mouth；The gas outlet 5 is positioned close to the side of culture bag bottom surface；The air inlet 4
It is arranged on sealing cover 3, the inoculation mouth 2 is spiral mouth, and the sealing cover 3 is helixseal lid, and sealing cover 3 is internally provided with
Silica gel ring, to ensure the sealing of 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 operation of the bubble in culture bag 1 to promote gas presentation to circulate state rather than vertically upward
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 gas outlet 5 is provided with
Filtering cotton 9 is for preventing miscellaneous bacteria from entering culture bag 1.In addition, the gas outlet 5 separates branch pipe(tube) as material-feeding port 6, for mending
Material.
Its application method is as follows：It is closed with devices such as tongs at the gas outlet of 1 bottom of culture bag when sterilizing, inoculation mouth 2
It is sealed using sterile sealed membrane, the sealing cover 3 and air inlet pipe of inoculation mouth 2, air sparger 7 and air disinfector, even
It is connected to together, then carries out whole sterilizing；After sterilizing, culture bag 1 is positive to place, and closes gas outlet, culture medium is injected
In culture bag 1, it is introduced directly into culture medium from inoculation mouth 2 after then shaking up a certain amount of cell together with culture solution；Then it is inoculated with
Mouth 2 seals, and is hung over culture bag on culturing rack using handle, keeps inoculation mouth 2 downward, and gas outlet 5 is upward, and connection intake pipe is beaten
The tongs on tracheae that begin to rehearse can ventilate, and make bubble and the culture solution that reacts in culture bag using the uphill process of bubble
It comes into full contact with, according to oneself needing to adjust ventilatory capacity size when use.In seeded process, it is big to cultivate culture medium height in bucket
In 20cm；In incubation, it is greater than 3cm between culture medium liquid level and gas outlet.
After culture, culture is poured out from inoculation mouth 2.
The culture bag product of the present invention is 2-100L, easy to operate, operates repeated strong, of low cost, dangerous small, microbiological contamination
Rate is low；The materials'use transparent material of culture bag and key fitment, can integrally sterilize, and culture medium capacity is stablized, and incubation is straight
It sees visible；Air inlet is equipped with air sparger and is obliquely installed, and carries out rotary air inlet, and training can be just met under low pass tolerance
The needs of base dissolved oxygen are supported, the loss of gas supply cost and culture medium is reduced.
Claims (10)
1. a kind of plant cell and organ Liquid Culture bag, which is characterized in that including culture bag and be arranged in culture bag bottom surface one
The handle at end is arranged in the gas outlet of culture bag bottom end, and inoculation mouth and air inlet in the culture bag other end is arranged；Its
In, one end of the culture bag setting inoculation mouth is wedge shape, inoculation mouth fitted seal lid.
2. culture bag as described in claim 1, which is characterized in that the culture bag uses transparent material.
3. culture bag as claimed in claim 1 or 2, which is characterized in that the gas outlet is positioned close to the side of bottom surface；Institute
It states air inlet to be arranged on the inclined-plane of wedge-shaped culture bag or be arranged on sealing cover, the air inlet connects air inlet pipe, will cultivate
Required gas imports in culture bag.
4. culture bag as claimed in claim 1 or 2, which is characterized in that a diameter of 2-20cm of inoculation mouth.
5. culture bag as claimed in claim 3, which is characterized in that the air inlet is arranged using inclined mode, it is described into
Gas port is provided with air sparger.
6. culture bag as claimed in claim 3, which is characterized in that the air inlet is arranged on sealing cover, the inoculation mouth
For spiral mouth, the sealing cover is helixseal lid, and sealing cover is internally provided with silica gel ring, to ensure the sealing of sealing cover.
7. culture bag as described in claim 1, which is characterized in that the angle of the wedge shape of the culture bag is 60-150 DEG C.
8. culture bag as claimed in claim 3, which is characterized in that the culture bag gas outlet connection is one or more
Opening, or it is additionally provided with one or more opening in the side close to gas outlet, the opening is used for feed supplement, connection
One or more feed supplement pump.
9. culture bag as claimed in claim 8, which is characterized in that the opening diameter is 0.2-4cm.
10. culture bag as described in claim 1, which is characterized in that gas filtration is arranged at the gas access of the air inlet pipe
The outlet of sterilization device, the gas outlet is provided with filtration sterilization component；The capacity of the culture bag is 2-100L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810398091.7A CN108441425A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ culture device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810398091.7A CN108441425A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ culture device |
Publications (1)
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| CN108441425A true CN108441425A (en) | 2018-08-24 |
Family
ID=63202223
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810398091.7A Pending CN108441425A (en) | 2018-04-28 | 2018-04-28 | A kind of plant cell, organ culture device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112430539A (en) * | 2019-08-26 | 2021-03-02 | 苏州君盟生物医药科技有限公司 | Bioreactor and application thereof |
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| JP2005040061A (en) * | 2003-07-23 | 2005-02-17 | Satoshi Takeuchi | Fermentation tank (jar fermenter) |
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Address after: 116000 Changdianbao Community, Pulandian Economic Development Zone, Dalian, Liaoning Province Applicant after: DALIAN PRACTICAL BIOTECHNOLOGY Co.,Ltd. Address before: 116023 No. 32 Lixian Street, Qixianling, Dalian High-tech Industrial Park, Liaoning Province Applicant before: DALIAN PRACTICAL BIOTECHNOLOGY Co.,Ltd. |
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Application publication date: 20180824 |