CN1320097C - Culturing system of built-in light biological reactor of plant tissue cutter - Google Patents

Culturing system of built-in light biological reactor of plant tissue cutter Download PDF

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Publication number
CN1320097C
CN1320097C CNB2005100232851A CN200510023285A CN1320097C CN 1320097 C CN1320097 C CN 1320097C CN B2005100232851 A CNB2005100232851 A CN B2005100232851A CN 200510023285 A CN200510023285 A CN 200510023285A CN 1320097 C CN1320097 C CN 1320097C
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coupling
cultivate
gas
cultivate system
built
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CN1680540A (en
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张宝红
齐瀚实
陈思晔
赵锐
黄娜
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Shanghai Jiaotong University
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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
    • C12M21/02—Photobioreactors
    • 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
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/02—Stirrer or mobile mixing elements
    • 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
    • C12M31/00—Means for providing, directing, scattering or concentrating light
    • C12M31/02—Means for providing, directing, scattering or concentrating light located outside the reactor
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/06—Means for regulation, monitoring, measurement or control, e.g. flow regulation of illumination

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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Abstract

The present invention relates to a cultivating system of a built-in photobiologic reactor of a plant tissue cutter, which comprises a liquid transferring system, a gas transferring system, a light irradiating system, a coupling cultivation system, a controller and an on-line light-sensitive monitor, wherein the liquid transferring system is connected with the coupling cultivation system by silicone rubber pipes, the gas transferring system is connected with the coupling cultivation system by silicone rubber pipes, the light irradiating system is fixed on the periphery of the coupling cultivation system, the on-line light-sensitive monitor is arranged on one side of the coupling cultivation system and is used for transmitting signals to the controller, and the controller receives electric optical signals from the on-line light-sensitive monitor, and is connected with the coupling cultivation system by a data wire. The present invention has the advantages that plant cells and plant tissues can be cut in an on-line mode in the cultivating process, and the metabolic growth state of the cultivated substances in the cultivating process is kept at an optimized point, and the polluting rate of the off-line cut is greatly reduced.

Description

The built-in light biological reactor of plant tissue cutter culture systems
Technical field
The present invention relates to a kind of culture systems, specifically is a kind of built-in light biological reactor of plant tissue cutter culture systems.Be used for plant tissue engineering field.
Background technology
The research of bioreactor starts from the large scale culturing of little algae forties in 20th century, developed rapidly after the eighties, developed miscellaneous bioreactor up to now, and can in bioreactor, insert pH value electrode, dissolved oxygen electrode, temperature electrode, cell density electrode, viscosity electrode etc., the signal that the sensor obtains can further be changed, realize the sequencing record and the control automatically of bioreactor.Land plant that is used to cultivate and large-scale seaweed cell or to organize mainly be forms such as callus, micropropagation body or gametophyte cell, its common feature is to have very strong growth, amplification ability, but continued growth after the cutting, and, individuality is constantly grown up in the culturing process, must smash just by regular cutting and can keep its undifferentiated state.The research of cultivating for the bioreactor of present plant tissue, in the culturing process when the tissue of cultivating and cell grow into certain volume and begin to influence its growth, because of present design does not all have at wire-electrode cutting device, cutting is smashed and is then wanted complete off-line operation as need, influence the experiment process, make troubles to experimental implementation, therefore, under the prerequisite that can not satisfy the demand on the hardware, the continuously long-term bioreactor culture apparatus system of online cutting can further realize the sequencing record and the control automatically of bioreactor in original design.
Find by prior art documents, people such as Yao-ming Huang are at " BiotechnologyProgress " 2002,18:62-71 has " the Dynamics of oxygen evolution and biomassproduction during cultivation of Agardhiella subulata microplantlrts ina bubble-column photobioreactor under medium perfusion " that deliver and (utilizes airlift photobioreactor to cultivate and visit the research that formula new Asia algae micropropagation body carries out oxygen transferring power and biomass, " biotechnology progress ") in the literary composition, the size that proposes the suitable suspension culture of Agardhiella subulata micropropagation body is 3-10mm, must regularly cut in the micropropagation body culturing process and smash, in order to keep the high growth rates of micropropagation body, when the micropropagation bulk-growth surpasses 10mm, be cut to the suitable size of 3-5mm again and continue again to cultivate in the culturing process; The culturing system of light bioreactor of the Agardhiella subulata micropropagation body of this article proposition simultaneously is made up of gas transmission system, illumination system and reactor body three parts.Its weak point is: this system can only cultivate for the micropropagation body provides interim, can not realize the cultivation of continuously long-term bioreactor, in addition, this system can only carry out the off-line cutting to the micropropagation body, this off-line cutting mode has increased the microbial contamination rate of culturing process and loaded down with trivial details cutting work greatly, makes troubles to experiment; When being applied to large scale culturing, the off-line cutting then becomes conditioning step, has a strong impact on carrying out smoothly of experiment.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, provide the online cutting of a kind of plant tissue continuously long-term bioreactor culture apparatus system, achieve the online cutting in vegetable cell and the tissue culture procedures, the growth metabolism state that guarantees culture in the culturing process maintains the optimization point, greatly reduces the pollution rate of off-line cutting simultaneously.
The present invention is achieved by the following technical solutions, the present invention includes: transfusion system, gas delivery system, illumination system, coupling cultivate system, controller, on-line photosensitive monitor.Its annexation is: transfusion system is connected with coupling cultivate system by silicone tube; Gas delivery system is connected with coupling cultivate system by silicone tube; Illumination system is fixed on the coupling cultivate system periphery; On-line photosensitive monitor is installed in coupling cultivate system one side, transmits signals to controller, and controller receives the photosignal from on-line photosensitive monitor, is connected with coupling cultivate system by data line.
Transfusion system is made up of container for storing liquid and infusion pump.Container for storing liquid is connected with coupling cultivate system through infusion pump by silicone tube, and the fresh medium in the container for storing liquid is in infusion pump flows into coupling cultivate system.
Gas delivery system is by air pump, first air filter, and gas meter, the gas humidification device is formed.The gas humidification apparatus main body is a cylinder shape hemisphere top Glass Containers, and its top is integrated with gas inlet and pneumatic outlet, and the gas inlet stretches to apart from about 1/6 place of container bottom; Be connected in turn by the silicone tube head and the tail between the gas inlet of air pump, first air filter, gas meter and gas humidifying device; The pneumatic outlet of gas humidification device is connected with gas distributor.
Coupling cultivate system is that bioreactor culture system and diced system are coupled, by fluid inlet, and gas distributor, dissolved oxygen electrode, pH electrode, whipping appts, stopple coupon, air outlet, second air filter, liquid outlet filters bolting silk, cutting unit, and the coupling cultivate system control panel is formed.Gas distributor is the hollow tube of being made by stainless material, form by ventpipe and gas distribution tube, ventpipe is inserted into the coupling cultivate system bottom position by the coupling cultivate system top, the gas distributor bottom is a gas distribution tube, be circular arc, be distributed in the periphery of coupling cultivate system rounded bottom surface, be integrated with circular production well above, ventpipe and gas distribution tube junction are the ventpipe contact; Whipping appts is made up of agitator arm and stirring rake, agitator arm by coupling cultivate system center of top position stretch into coupling cultivate system inside far from the bottom 1/3～1/4 place, agitator arm bottom is integrated with stirring rake; Stopple coupon stretch into coupling cultivate system inside far from the bottom about 1/2 place; Dissolved oxygen electrode and pH electrode stretch under the liquid level of coupling cultivate system; The place, air outlet connects second air filter with silicone tube; The liquid outlet place has double-deck 800 purposes to filter bolting silk.
Transfusion system inputs to coupling cultivate system with fresh medium by fluid inlet, and for keeping constant volume, excessive nutrient solution is flowed out automatically by the liquid outlet place in the coupling cultivate system, and there is the filtration bolting silk at the liquid outlet place; Gas delivery system links to each other with gas distributor by silicone tube, and gas distributor is inserted into the coupling cultivate system bottom position by the coupling cultivate system top, and for its cultivation provides competent gas, residual gas and waste gas are discharged through the air outlet; Dissolved oxygen electrode and pH electrode stretch into coupling cultivate system inside by the coupling cultivate system top; Cutting unit is incorporated on the coupling cultivate system control panel base, is positioned at middle position, is the cross knife-edge, and two ends are inclined upwardly and erect; Be integrated with the cutting parameter setting knob on the coupling cultivate system control panel.
When the present invention works, fresh medium in the container for storing liquid is in infusion pump flows into coupling cultivate system, to keep the long-term cultured continuously of coupling cultivate system, constant for the volume of culture that keeps coupling cultivate system, excessive nutrient solution is flowed out automatically by the liquid outlet place, the gas that air pump provides is through the first air filter filtration sterilization, behind the gas humidification device humidification, provide competent gas in the coupling cultivate system bottom for its cultivation by gas distributor, remaining gas and waste gas are discharged by the air outlet, the place, air outlet connects second air filter, to guarantee sterile state in the reactor, illumination system provides necessary light intensity for the coupling mechanism culture systems, coupling cultivate system enters cultivation stage, the online photosensitive watch-dog of coupling cultivate system one side is determined the size of purpose culture by photosensor signal, continuously, on-line monitoring purpose culture size and concentration, and with signal feedback to controller, the size of purpose culture reaches the setting size parameter in feedback information, when simultaneously quantity reaches preset proportion and counts, controller is started working, controller is directly controlled the cutting unit switch of integrating on the coupling cultivate system control panel, start cutting unit, the cutting beginning, when the size of purpose culture in the feedback information and quantity are lower than setup parameter, controller cuts out the startup of cutting unit, cutting stops voluntarily, cutting finishes, and coupling cultivate system enters cultivation stage.
System of the present invention is a closed system; reduced microbiological contamination to greatest extent; solved simultaneously in vegetable cell and the tissue culture procedures because of the individual excessive practical problems of bringing to experiment of purpose culture; guaranteed that the growth metabolism state of culture maintains the optimization point in the culturing process; realized the continuously long-term bioreactor cultivation of the online cutting of plant tissue, for the possible approach that provides is provided in mass-producing.
Compared with prior art, the present invention combines use with cutting unit and bioreactor, realized the online cutting in vegetable cell and the tissue culture procedures, improved working efficiency, guaranteed that the quality and the growth metabolism state thereof of purpose culture maintains the optimization point in the culturing process; Control panel is cultivated in coupling among the present invention, and is workable, can set different cutting parameters according to different purpose cultures; Replace traditional visual method by on-line photosensitive monitor testing goal culture size, improved science, the accuracy of experiment; The cutting unit culturing system of built-in light biological reactor is applicable to the macro-organism reactor, and the system height intensification can be controlled automatically, and floor space is little, and is easy and simple to handle, is easy to control, reduced the systematic error of bringing because of cutting to greatest extent.
Description of drawings
Fig. 1 is a structural representation of the present invention
Fig. 2 is a transfusion system structural representation of the present invention
Fig. 3 is a gas delivery system structural representation of the present invention
Fig. 4 is a coupling cultivate system structural representation of the present invention
Fig. 5 is a gas distributor structure front view of the present invention
Fig. 6 is a gas distributor structure vertical view of the present invention
Among the figure, 1. 14. gas vents, 15. inlets, 16. gas distributors, 17. dissolved oxygen electrode 18.pH electrodes, 19. agitating devices, 20. probe tubes, 21. gas outlets, transfusion system 2. gas delivery systems 3. illumination systems 4. coupling cultivate systems 5. controllers 6. on-line photosensitive monitors, 7. fluid reservoirs, 8. infusion pumps, 9. air pumps, 10. first air cleaners, 11. gas flowmeters, 12. gas humidification devices, 13. gas accesses, 22. second air cleaners, 23. liquid outlets 24. filter bolting silk 25. cutters 26. coupling cultivate system control panels 27. puddlers 28. paddles 29. breather pipes 30. gas distribution tubes 31. breather pipe contacts 32. ventholes
Embodiment
Shown in Fig. 1-6, the present invention includes: transfusion system 1, gas delivery system 2, illumination system 3, coupling cultivate system 4, controller 5, on-line photosensitive monitor 6.Its annexation is: transfusion system 1 is connected with coupling cultivate system 4 by silicone tube; Gas delivery system 2 is connected with coupling cultivate system 4 by silicone tube; Illumination system 3 is fixed on coupling cultivate system 4 peripheries; On-line photosensitive monitor 6 is installed in a side of coupling cultivate system 4, transmits signals to controller 5, and the photosignal that controller 5 receives from on-line photosensitive monitor 6 is connected with coupling cultivate system 4 by data line.
Transfusion system 1 is made up of container for storing liquid 7 and infusion pump 8.Container for storing liquid 7 is connected with coupling cultivate system 4 through infusion pump 8 by silicone tube, and the fresh medium in the container for storing liquid 7 flows in the coupling cultivate system 4 through infusion pump 8.
Gas delivery system 2 is by air pump 9, the first air filters 10, gas meter 11, and gas humidification device 12 is formed.The top of gas humidification device 12 is integrated with gas inlet 13 and pneumatic outlet 14, and gas inlet 13 stretches to apart from about 1/6 place of container bottom; Connect in turn by the silicone tube head and the tail between the gas inlet 13 of air pump 9, first air filter 10, gas meter 11 and gas humidifying device 12; The pneumatic outlet 14 of gas humidification device 12 is connected with coupling cultivate system 4.
Coupling cultivate system 4 is that bioreactor culture system and diced system are coupled, by fluid inlet 15, and gas distributor 16, dissolved oxygen electrode 17, pH electrode 18, whipping appts 19, stopple coupon 20, air outlet 21, second air filter 22, liquid outlet 23 filters bolting silk 24, cutting unit 25, coupling cultivate system control panel 26 is formed.Gas distributor 16 is hollow tubes of being made by stainless material, form by ventpipe 29 and gas distribution tube 30, ventpipe 29 is inserted into coupling cultivate system 4 bottom positions by coupling cultivate system 4 tops, gas distributor 16 bottoms are gas distribution tube 30, be circular arc, be distributed in the periphery of coupling cultivate system 4 rounded bottom surfaces, be integrated with circular production well 32 above, ventpipe 29 and gas distribution tube 30 junctions are ventpipe contact 31; Whipping appts 19 is made up of agitator arm 27 and stirring rake 28, agitator arm 27 by coupling cultivate system 4 center of top positions stretch into coupling cultivate system 4 inside far from the bottom 1/3～1/4 place, agitator arm 27 bottoms are integrated with stirring rake 28; Stopple coupon 20 stretch into coupling cultivate system 4 inside far from the bottom about 1/2 place; Dissolved oxygen electrode 17 and pH electrode 18 stretch under the liquid level of coupling cultivate system 4; 21 places, air outlet connect second air filter 22 with silicone tube; Transfusion system 1 inputs to coupling cultivate system 4 with fresh medium by fluid inlet 15, and excessive nutrient solution is flowed out automatically by liquid outlet 23 places in the coupling cultivate system 4, and liquid outlet 23 places have double-deck 800 purposes to filter bolting silk 24; Cutting unit 25 is incorporated on culture systems control panel 26 bases, is positioned at middle position, is the cross knife-edge, and two ends are inclined upwardly and erect; Be integrated with the cutting parameter setting knob on the coupling cultivate system control panel 26.

Claims (5)

1, a kind of built-in light biological reactor of plant tissue cutter culture systems, comprise: transfusion system (1), gas delivery system (2), illumination system (3), it is characterized in that, also comprise: coupling cultivate system (4), controller (5), on-line photosensitive monitor (6), its mode of connection is: transfusion system (1) is connected with coupling cultivate system (4) by silicone tube, gas delivery system (2) is connected with coupling cultivate system (4) by silicone tube, illumination system (3) is fixed on coupling cultivate system (4) periphery, on-line photosensitive monitor (6) is arranged on a side of coupling cultivate system (4), transmit signals to controller (5), be connected with coupling cultivate system (4) by data line;
Coupling cultivate system (4) is by fluid inlet (15), gas distributor (16), dissolved oxygen electrode (17), pH electrode (18), whipping appts (19), stopple coupon (20), air outlet (21), second air filter (22), liquid outlet (23), filter bolting silk (24), cutting unit (25), coupling cultivate system control panel (26) is formed, gas distributor (16) is distributed in the periphery of coupling cultivate system (4) rounded bottom surface, whipping appts (19) is made up of agitator arm (27) and stirring rake (28), agitator arm (27) by coupling cultivate system (4) center of top position stretch into coupling cultivate system (4) inner apart from the bottom 1/3～1/4 place, agitator arm (27) bottom is integrated with stirring rake (28), stopple coupon (20) stretch into coupling cultivate system (4) inner apart from the bottom 1/2 place, dissolved oxygen electrode (17) and pH electrode (18) stretch under the liquid level of coupling cultivate system (4), air outlet (21) is located to connect second air filter (22) with silicone tube, transfusion system (1) inputs to coupling cultivate system (4) with fresh medium by fluid inlet (15), excessive nutrient solution is located automatic outflow by liquid outlet (23) in the coupling cultivate system (4), liquid outlet (23) has been located double medium filtration bolting silk (24), cutting unit (25) is incorporated on culture systems control panel (26) base, and the coupling cultivate system control panel is integrated with the cutting parameter setting knob on (26).
2, built-in light biological reactor of plant tissue cutter culture systems according to claim 1, it is characterized in that, transfusion system (1) is made up of container for storing liquid (7) and infusion pump (8), and container for storing liquid (7) is connected with coupling cultivate system (4) through infusion pump (8) by silicone tube.
3, built-in light biological reactor of plant tissue cutter culture systems according to claim 1, it is characterized in that, gas delivery system (2) is by air pump (9), first air filter (10), gas meter (11), gas humidification device (12) is formed, the top of gas humidification device (12) is integrated with gas inlet (13) and pneumatic outlet (14), gas inlet (13) stretches to apart from about 1/6 place of container bottom, air pump (9), first air filter (10), connected in turn by the silicone tube head and the tail between the gas inlet (13) of gas meter (11) and gas humidifying device (12), the pneumatic outlet (14) of gas humidification device (12) is connected with coupling cultivate system (4).
4, built-in light biological reactor of plant tissue cutter culture systems according to claim 1, it is characterized in that, gas distributor (16) is the hollow tube of being made by stainless material, form by ventpipe (29) and gas distribution tube (30), ventpipe (29) links to each other with the pneumatic outlet (14) of gas humidification device (12), be inserted into coupling cultivate system (4) bottom position by coupling cultivate system (4) top, gas distributor (16) bottom is gas distribution tube (30), be circular arc, be distributed in the periphery of coupling cultivate system (4) rounded bottom surface, be integrated with circular production well (32) above, ventpipe (29) and gas distribution tube (30) junction are ventpipe contact (31).
5, built-in light biological reactor of plant tissue cutter culture systems according to claim 1, it is characterized in that cutting unit (25) is incorporated on coupling cultivate system control panel (26) base, is positioned at middle position, be the cross knife-edge, two ends are inclined upwardly and erect.
CNB2005100232851A 2005-01-13 2005-01-13 Culturing system of built-in light biological reactor of plant tissue cutter Expired - Fee Related CN1320097C (en)

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CN108522247A (en) * 2018-04-28 2018-09-14 刘汉石 A kind of bioreactor of culture ginseng adventitious root
CN111528103B (en) * 2020-07-01 2021-10-08 广州齐志生物工程设备有限公司 Plant tissue culture reactor and amplification culture method thereof
CN117256479B (en) * 2023-11-22 2024-02-06 上海科缔联科技有限公司 Industrial automatic biological reaction device and control method thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1201826A (en) * 1997-06-05 1998-12-16 华南师范大学 Spirulina light illuminating tower-tray optic bio-reactor culturing system and its controlling method
CN1560222A (en) * 2004-03-04 2005-01-05 江南大学 Coupling biologic reactor

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1201826A (en) * 1997-06-05 1998-12-16 华南师范大学 Spirulina light illuminating tower-tray optic bio-reactor culturing system and its controlling method
CN1560222A (en) * 2004-03-04 2005-01-05 江南大学 Coupling biologic reactor

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