CN208857315U - A kind of plant cell and organ Liquid Culture bag - Google Patents

A kind of plant cell and organ Liquid Culture bag Download PDF

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Publication number
CN208857315U
CN208857315U CN201820626195.4U CN201820626195U CN208857315U CN 208857315 U CN208857315 U CN 208857315U CN 201820626195 U CN201820626195 U CN 201820626195U CN 208857315 U CN208857315 U CN 208857315U
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China
Prior art keywords
culture bag
culture
air inlet
bag
inoculation mouth
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CN201820626195.4U
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刘汉石
吴俊罡
金银兵
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PURUIKANG BIOTECHNOLOGY CO Ltd DALIAN
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PURUIKANG BIOTECHNOLOGY CO Ltd DALIAN
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Abstract

The utility model provides a kind of easy plant cell and organ Liquid Culture bag and the gas outlet of culture bag bottom end is arranged in including culture bag and the handle that culture bag bottom end is arranged in, and the inoculation mouth and air inlet of the culture bag other end is arranged in;Wherein, one end of the culture bag setting inoculation mouth is wedge shape, and inoculation mouth cooperates sealing cover.Compared with existing culture bag, the culture bag product of the utility model is 2-100L, easy to operate, operates repeatability by force, low in cost, risk is small, and 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

A kind of plant cell and organ Liquid Culture bag
Technical field
The utility model relates to technical field of cell culture, and in particular to a kind of a kind of plant of suitable large-scale production is thin Born of the same parents, organ Liquid Culture bag.
Background technique
For plant available resources, China is one of the most abundant country of resources of medicinal plant 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 mainly includes stirring Formula reactor, airlift reactor;The latter mainly includes 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, serfin [A.H.Scragg et al., Enzyme Microb is produced with airlift bioreactor culture Catharanthus Roseus Cell Technol, 1987,9:619];South Korea S.H.Son et al. to yew cell carry out reactor large-scale culture, using from The airlift reactor [S.H.Son et al., Plant Cell Reports (2000) 19:628-633] of 100L to 500L; After several years, Kee-Yoeup Paek carries out large-scale culture to ginseng adventitious root using spherical bubbling style reactor and obtains 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's using gas is followed 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 Usually 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 have the following problems: 1) currently used reactor is all to be fixed on 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 into using stainless steel as material, therefore in production It requires setting 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 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 not can be carried out sterilizing, the type is set It needs steam-out (SO) to sterilize when standby sterilizing, pays particular attention to the risk and complexity of high steam, and steam goes out Bacterium sterilizes while will lead to culture volume increase, and it is unstable to accelerate, and also results in substance volatile in culture medium And gasification temperature is discharged lower 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 Liquid phase contact stirs liquid to improve mass transfer rate while reaction.However the advantages of being above theoretically, really Realize 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 easy to appear without agitating device It is mixed serious, bubble be also easy to produce it is poly- simultaneously, and phenomena such as 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 ventilates and will lead to a large amount of evaporations of culture medium, generates not to the growth of plant cell Good influence, and efficiency is lower, although can weaken this influence by the way that defoaming agent is added, defoaming 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 ventilatory capacity is excessive leads to some materials for needing long-term cultivation Material, later period can have to terminate culture because of the evaporation of culture medium.Many equipment and stirring type bioreactor one of the type Sample needs to sterilize using external high steam, increases risk and complexity when operation, and culture medium holds when sterilizing The unstability of amount.
Complicated for operation to solve equipment valuableness 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, this is practical It is novel it is quasi- provide can meet dissolved oxygen demand under a kind of low pass gas, feed supplement equipment can be sterilized simultaneously together with culture device, be cultivated The lower incubator of bubble formation rate in the process.
Utility model content
The purpose of the utility model is to provide a kind of plant cells and organ Liquid Culture bag.
In order to solve the above technical problems, the utility model adopts the following technical solution:
A kind of plant cell and organ Liquid Culture bag, including culture bag and the handle that culture bag bottom surface one end is arranged in, The gas outlet of culture bag bottom end is set, and the inoculation mouth and air inlet of the culture bag other end are set;Wherein, the training The one end for supporting bag setting inoculation mouth is wedge shape, and inoculation mouth cooperates sealing cover.
The culture bag of the utility model is made of transparent material, can observe the life of culture during the cultivation process in this way Long status, if 121 DEG C of high pressure sterilizations can be carried out using heat-resisting material, if can be with using the non-refractories such as PE material Irradiation sterilization is carried out, in sterilizing, can integrally be sterilized.
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 culture medium after shaking up from inoculation mouth;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 below, and gas outlet is located above, and then passes to gas, utilizes the rising of bubble Journey comes into full contact with bubble 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 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, increase the gas out of plumb being discharged into;
Further, the air inlet is additionally provided with air sparger, to promote gas presentation to circulate state It is non-perpendicular upward, extend running track 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 guarantee 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 2-10cm.
Further, the angle of the wedge shape of the culture bag is between 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;During the cultivation process, culture medium liquid level with go out It is greater than 3cm between 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, the outlet of the gas outlet is provided with filtration sterilization component to prevent living contaminants, 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 at the gas outlet of bottom with devices such as tongs, 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, is connected 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 utility model is 2-100L, and culture bag can fold, be highly convenient for operating.
The purpose of the utility model is to provide a kind of easy plant cells and organ Liquid Culture bag, and main advantage is such as Under:
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, can both find to handle as early as possible early 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 rotate along bucket wall, promote gas between culture medium and bubble Exchange is realized and meets the needs of culture medium dissolved oxygen under low pass tolerance, therefore can not be seldom or even in practical incubation Use defoaming 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 is evaporated 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 placed on aseptic operating platform, to carry out, and greatly reduces and connects Contamination rate when kind;Operating process is simple and convenient, and risk is small, and contamination rate is low, operates repeatability by force, low in cost, can be significantly Reduce the production cost of culture.
Detailed description of the invention
Fig. 1 is the schematic diagram of plant cell and organ Liquid Culture bag described in the utility model embodiment 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 embodiment
Plant cell described in the utility model and organ Liquid Culture bag are further described with reference to the accompanying drawing.
Embodiment 1
A kind of plant cell and organ Liquid Culture bag, including culture bag 1 (by can PE be made, capacity 40L), including Culture bag 1 and the handle 10 that 1 bottom surface one end of culture bag is set, and the inoculation mouth 2 and air inlet of 1 other end of culture bag are set Mouth 4,2 diameter of inoculation mouth are 10cm;The gas outlet 5 of 1 bottom end of culture bag is set;Wherein, the setting of culture bag 1 inoculation One end of mouth 2 is wedge shape, and the angle of the wedge shape of the culture bag 1 is 64 DEG C, and inoculation mouth 2 is arranged in wedge-shaped top, inoculation mouth 2 Height is 12cm, and inoculation mouth 2 cooperates sealing cover 3;The gas outlet 5 is positioned close to the side of culture bag bottom surface;The air inlet Mouth 4 is arranged on sealing cover 3, and the inoculation mouth 2 is spiral mouth, and the sealing cover 3 is helixseal lid, sets inside sealing cover 3 It is equipped with silica gel ring, to guarantee the sealing of sealing cover 3.
Air inlet 4 is arranged using inclined mode, increase the gas out of plumb being discharged into;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 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: being closed at the gas outlet of 1 bottom of culture bag with devices such as tongs 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 be placed, and closes gas outlet, culture medium is injected In culture bag 1, it is introduced directly into culture medium after then shaking up a certain amount of cell together with culture solution from inoculation mouth 2;Then it is inoculated with Mouth 2 seals, and is hung over culture bag on culturing rack using handle, makes inoculation mouth 2 downwards, and upwards, connection intake pipe is beaten for gas outlet 5 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;During the cultivation process, 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 utility model is 2-100L, easy to operate, operates repeatability by force, and low in cost, risk is small, 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 cultivate Cheng Zhiguan is visible;Air inlet is equipped with air sparger and is obliquely installed, and carries out rotary air inlet, can be just full under low pass tolerance The needs of sufficient culture medium dissolved oxygen reduce the loss of gas supply cost and culture medium.

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 the inoculation mouth and air inlet of the culture bag other end is arranged in;Its In, one end of the culture bag setting inoculation mouth is wedge shape, and inoculation mouth cooperates sealing cover.
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 the inoculation mouth diameter is 2-10cm.
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 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 guarantee 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 Sterilization device, the outlet of the gas outlet are provided with filtration sterilization component;The capacity of the culture bag is 2-100L.
CN201820626195.4U 2018-04-28 2018-04-28 A kind of plant cell and organ Liquid Culture bag Active CN208857315U (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021103863A1 (en) 2019-11-28 2021-06-03 大连普瑞康生物技术有限公司 Method for increasing the content of flavonoid phenylpropanoid compounds in saussurea involucrata cell culture

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021103863A1 (en) 2019-11-28 2021-06-03 大连普瑞康生物技术有限公司 Method for increasing the content of flavonoid phenylpropanoid compounds in saussurea involucrata cell culture

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GR01 Patent grant
GR01 Patent grant
PE01 Entry into force of the registration of the contract for pledge of patent right
PE01 Entry into force of the registration of the contract for pledge of patent right

Denomination of utility model: A liquid culture bag for plant cells and organs

Effective date of registration: 20200930

Granted publication date: 20190514

Pledgee: Dalian Xigang District Enterprise Credit Financing Guarantee Co., Ltd

Pledgor: DALIAN PRACTICAL BIOTECHNOLOGY Co.,Ltd.

Registration number: Y2020210000054

PC01 Cancellation of the registration of the contract for pledge of patent right
PC01 Cancellation of the registration of the contract for pledge of patent right

Date of cancellation: 20210929

Granted publication date: 20190514

Pledgee: Dalian Xigang District Enterprise Credit Financing Guarantee Co., Ltd

Pledgor: DALIAN PRACTICAL BIOTECHNOLOGY Co.,Ltd.

Registration number: Y2020210000054