CN1648240A - Periodic immersion air lifting type plant cell cultivating method and its cultivating device - Google Patents
Periodic immersion air lifting type plant cell cultivating method and its cultivating device Download PDFInfo
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- CN1648240A CN1648240A CN 200410000797 CN200410000797A CN1648240A CN 1648240 A CN1648240 A CN 1648240A CN 200410000797 CN200410000797 CN 200410000797 CN 200410000797 A CN200410000797 A CN 200410000797A CN 1648240 A CN1648240 A CN 1648240A
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Abstract
The present invention relates to periodically immersing airlift type plant cell cultivating method and apparatus. After plant cell is set on multilayer turnable sieve plates inside a reactor, the sieve plates are turned to make the cell in immersed suspension cultivation stage and no-immersion solid cultivation stage alternately until completing cultivation. There are pull rods to fix the sieve plates onto the electromagnetic pull rod of electromagnetic pull rod device, time control instrument to control the period turning of the sieve plates, and solenoid valve to complete the alternate immersion and no immersion of culture liquid to cell, ventilation and lighting. The alternate cell immobilizing and suspending culture can regulate the cell aggregation body size effectively and this is favorable to biological information and nutritious matter transfer among cells, and suitable for cell growth and metabolism and possesses excellent mixing, mass transfer and heat transfer characteristic.
Description
Technical field
Particularly a kind of culture plant cell method and culture apparatus thereof of cycle submergence air lift type are cultivated in the mass-producing that the present invention relates to use bio-reactor to carry out vegetable cell in the biological technical field.
Background technology
Vegetable cell, tissue, organ large scale culturing technology are effective ways of producing useful secondary metabolite, have obtained very big development in recent years.The bio-reactor that is used for vegetable cell, tissue, organ culture is reactor improvement, the development and next from the culturing micro-organisms cell, but characteristics with the uniqueness that adapts to plant cell growth, mainly be divided into mechanical stirring bio-reactor, on-mechanical stirred bioreactor, bubble tower and airlift bioreactor, packed bed bio-reactor, fluidized bed bio reactor, membrane bioreactor etc.Because the inherent nature of vegetable cell, assemble easily, easily break up as cell, the complicacy that the vulnerability of cell, pathways metabolism and meta-bolites form and cell is grown etc., the meta-bolites of selecting suitable reactor configuration and working method greatly to be related to vegetable cell synthesizes.Mitsui Petrochemical Ind., Ltd. as Japan adopts 750L stirring reactor two step method to cultivate paniculatum cell, realize suitability for industrialized production (the Noguchi et al. of shikonin, Plant tissue culture and itsbiotechnological application, 1977, pp.85.).Germany has set up the culture plant cell factory that 75L, 750L, 7500L, 15000L and 75000L series stirring type bioreactor are formed, and utilize these reactors to cultivate Echinacea purpurea Moench (Echinaceapurpurea) cell, production immunocompetence polysaccharide (Mavituna F, Recent advancesin biotechnology, 1992, pp.209-226).
Large scale culturing for plant tissue and organ, as: the bud of growing thickly, hairly root, body embryo and seedling etc., because these organs and tissue volume are big, form is special, mobile poor in culturing process, easily pile up, to more sensitivity and easier generation vitrifying of long period of soaking and variation etc. of shearing force, the scientific research personnel has used the bio-reactor of number of different types to carry out the cultivation of plant tissue and organ according to the different mining of Object of Development both at home and abroad, reported that as Rodrigueg-Mendiola etc. a kind of plant hairly root of 9 liters cultivates reactor (M.A.Rodrigueg-Mendiol et al., Bioreactors for growth of plant roots, Enzyme Microb.Technol.1991, Vol.13, pp.697-702), this reactor section is divided inoculation and the accumulation problem that has solved hairly root, but the not basic homogeneity that solves inoculation, thereby mixed characteristic and mass transfer have been influenced, heat transfer process, and this reactor is full immersion, easily causes the vitrifying and the variation of culture.At vegetable cell, tissue and organ growth metabolic characteristic, a kind of Airlift cyclic immersion lighting plant cell tissue organ of the applicant is cultivated reactor, and (on November 13rd, 1999 authorized, the patent No. is 98205797.0), this reactor is mainly by reactor body, the sieve plate dividing plate, inoculated tube, gas distributor, light source, reservoir, magnetic valve, gas-liquid circulation tube and gas-liquid distribution piping are formed, it is a kind of bio-reactor that is suitable for large scale culturing, can realize the even inoculation of plant tissue and organ culture, overcome vitrifying and variation phenomenon that the long-term submergence of plant tissue organ produces, culture is supported fully in the reactor, can also realize that gas-liquid replaced for half submergence cycle, the adjusting of periodicity of illumination and control, has good mixing, mass transfer, heat transfer characteristic, but, this bio-reactor is more suitable for the cultivation of tissue and the organ of plant, as the not runny bud of growing thickly, hairly root, seedling etc., for some vegetable cell that need control the cell aggregation size and need immobilization and suspension culture to combine, then have and to realize shortcoming fixing and that totally circulate simultaneously.
Summary of the invention
The objective of the invention is: overcome the cultivation that existing plant cell tissue organ culture reactor only is suitable for tissue and the organ of plant, can not control the shortcoming that the secondary metabolite of size, the existence of cell aggregation yields poorly, and pre-existing reactors complicated problems too, thereby provide a kind of culture plant cell method and reactor thereof of cycle submergence air lift type, to be suitable for culturing plants cell on a large scale.
Technical scheme of the present invention is as follows:
The cycle submergence air lift type cultural method of vegetable cell provided by the invention, its step is as follows:
1) vegetable cell is inserted on the rotatable sieve plate of multilayer in the transparent reactor main body after the sterilization;
2) plant cell cycle is alternately carried out nutrient solution full buried circulation suspension culture and the solid-state cultivation of the non-buried of nutrient solution, finish until cultivation;
The full buried circulation of described nutrient solution suspension culture is: be under the full buried state of nutrient solution at vegetable cell, rotating rotatable sieve plate makes it horizontal by 45 ° of-75 ° of angles, at this moment, vegetable cell on the sieve plate is suspended in the nutrient solution, carries out the full buried circulation of nutrient solution suspension culture 1-8 hour under the blowing from reactor body bottom gas distributor gas;
The solid-state cultivation of the non-buried of described nutrient solution is: sieve plate is rotated and when being fixed to level attitude, nutrient solution is put into the container for storing liquid that is arranged in its below from the bottom of reactor body, at this moment, vegetable cell drops to and carries out the solid-state cultivation of the non-buried of nutrient solution 10-60 minute on the rotatable sieve plate that is horizontal attitude;
The full buried circulation of the nutrient solution that described periodicity hockets suspends and the solid-state cultivation of the non-buried of nutrient solution is carried out under the illumination of the external light source that is positioned at incubator main body periphery, and its intensity of illumination is 1000-4000Lux, and light application time is 8-24 hour every day.
The cycle submergence air lift type culture apparatus of vegetable cell provided by the invention comprises:
The reactor body that one transparent material is made, fluid infusion mouth on it is connected with a fluid infusion jar by a valve, the reactor body inner bottom part is installed a gas distributor that communicates with source of the gas, the below of reactor body is provided with a container for storing liquid that communicates with the reactor body bottom, this container for storing liquid communicates with source of the gas, it is characterized in that, also comprise be installed in the reactor body can be at the sieve plate that in 45 ° of-75 ° of angular regions, rotates;
One sieve plate bracing frame also can be installed in the reactor body, and described sieve plate is fixed on the sieve plate bracing frame by back shaft, and sieve plate is rotating in 45 ° of-75 ° of angular regions around back shaft;
One end passes the pull bar of reactor body top cover and the outward flange of described sieve plate is fixed, and stretches out the outer tie rod end of reactor body and links to each other with the handle that pull bar is moved up and down; The described outer tie rod end of reactor body of stretching out links to each other with the electromagnetic draw bar device that pull bar is moved up and down, described electromagnetic draw bar device is sealed to form after placing an electromagnetic draw bar in the coil, the end and the electromagnetic draw bar of described pull bar are fixed together, described coil links to each other with a time controller, can realize that through time controller control the periodicity of multilayer sieve plate rotates automatically.The top of electromagnetic draw bar device is provided with vapor pipe, is beneficial to the heat radiation of electromagnetic draw bar main body and the quick elimination of outage back transient magnetic field, guarantees the quick falling of sieve plate;
One magnetic valve that is electrically connected with the time controller is installed on the connecting pipe between described container for storing liquid and the source of the gas, one magnetic valve that is electrically connected with the time controller is installed on the connecting pipe of container for storing liquid inlet mouth and reactor body air, two magnetic valves open/close opposite states, cultivate with the solid-state cultivation of non-submergence with the full immersion liquid of realizing nutrient solution and hocket;
Also can be evenly distributed with external light source around described reactor body, its intensity of illumination is 1000-4000Lux; Suit one thermostat(t)ed water chuck around described container for storing liquid is in order to the temperature of regulation and control nutrient solution; On described container for storing liquid, thief hole is set, can measures the pH of nutrient solution, various concentrations of nutrient at any time, the cultivation situation in the reactor body is monitored.
The invention has the advantages that: the culture plant cell method and the reactor thereof of cycle submergence air lift type of the present invention, can realize that many vegetable cells carry out periodicity submergence, ventilation, periodically illumination, be more suitable for the characteristics and the biorhythm of plant cell growth, the periodic rotation automatically of each layer sieve plate makes the cell cycle circulation, the cell that guarantees the submerged culture phase evenly is suspended in the whole reactor, cell obtains sufficient nutrition, and, the shear action that circulation time is suitable can suppress cell aggregation and excessively increase; The cell of non-submerged culture phase then can evenly be attached on each layer sieve plate, helps cell aggregation and grows up.What the present invention had realized cell fixation and the circulation that suspends is cultivated hockets, the pair cell aggregate can effectively be regulated and control, make the control of aggregate size within the specific limits, this helps the transmission that intercellular bioinformation transmission and nutritive substance comprise oxygen and carbon dioxide, this reactor helps the growth and the metabolism of cell, has good mixing, mass transfer, heat transfer characteristic.
Description of drawings
Fig. 1 is the structural representation of culture apparatus of the present invention;
When Fig. 2 cultivated vegetable cell for using culture apparatus of the present invention and cultural method, vegetable cell was in the cell aggregation flow schematic diagram of the full submergence circulation of nutrient solution suspension culture during the phase;
Fig. 3 is an electromagnetic draw bar apparatus structure synoptic diagram;
Wherein:
Reactor body 1 sieve plate frame 2 gas distributors 3
Inoculation mouthful 4 ventages, 5 pull bars 6
Vapor pipe 7 fluid infusion jars 8 airs 9
First magnetic valve, 10 container for storing liquids, 11 tubings 12
Container for storing liquid inlet mouth 13 thermostat(t)ed water chucks 14 membrane type air filter 15a
Membrane type air filter 15b second magnetic valve 16 gas meter 17a
Gas-holder 20 thief holes 21 external light sources 22
Embodiment
Embodiment 1:
The cycle submergence air lift type culture apparatus of the vegetable cell of the 2L volume of present embodiment, structure such as Fig. 1, as shown in Figure 1, culture apparatus comprises:
The reactor body 1 that one transparent material is made, fluid infusion mouth on it is connected with a fluid infusion jar 8 by a valve, reactor body 1 inner bottom part is installed a gas distributor 3 that communicates with source of the gas 20, the below of reactor body 1 is provided with a container for storing liquid 11 that communicates with the reactor body bottom, this container for storing liquid 11 communicates with source of the gas 20, in reactor body 1 can be at 2 layers of sieve plate that in 60 ° of scopes, rotate; Described sieve plate can be fixed on the sieve plate bracing frame 2 by back shaft, sieve plate bracing frame 2 usefulness silica gel sheets 25 is installed in the reactor body 1 again, and sieve plate is rotational fixation otherwise also;
One end passes the pull bar 6 of reactor body 1 top cover and fixes with the outward flange of described sieve plate, the end of stretching out the pull bar 6 outside the reactor body 1 links to each other with the electromagnetic draw bar device 28 that pull bar 6 is moved up and down, its structure is: stretch out reactor body 1 tie rod end outward and be fixed on the electromagnetic draw bar 27 that places in the coil 26, this coil 26 connects a time controller 23, also is provided with vapor pipe 7 at the top of electromagnetic draw bar device 28 main bodys;
One second magnetic valve 16 that is electrically connected with time controller 23 is installed on the connecting pipe between described container for storing liquid 11 and the source of the gas 20, what one first magnetic valve, 10, two magnetic valves that are electrically connected with the time controller were installed on the connecting pipe of container for storing liquid inlet mouth 13 and reactor body air 9 opens/closes opposite states;
Be evenly distributed with external light source 22 (tubular fluorescent lamp) around the described reactor body 1, its intensity of illumination is 1000Lux.Suit one thermostat(t)ed water chuck 14 around the described container for storing liquid 11;
Below reactor body 1, place container for storing liquid 11, and be communicated with the bottom of container for storing liquids 11 and reactor body 1 with a tubing 12, after the inlet mouth 13 at container for storing liquid 11 tops is connected on the membrane type air filter 15b by an inlet pipe, link to each other with inlet system with gas meter 17b by second magnetic valve 16, link to each other with ventage 9 on the reactor body blind flange by first magnetic valve 10 at another root ventpipe that picks out on inlet mouth 13 pipelines, two magnetic valves and coil 26 link to each other with time controller 23, rotate automatically with the submergence that realizes nutrient solution and the cycle of non-submergence alternate cycles and multilayer sieve plate.
Culture apparatus and cultural method of the present invention with embodiment 1 are cultivated vegetable cell (Saussurea medusa):
Under gnotobasis vegetable cell (Saussurea medusa) added on each layer sieve plate in the reactor body 1, and the full submergence circulation of the nutrient solution that hockets cultivates and the solid-state cultivation of non-submergence, its concrete operations step is as follows:
At first, coil 26 is taken off, again the whole reactor system is put in the steam sterilizing pot with the high-temperature steam sterilization, after the sterilization, treat that culture-liquid temp is reduced to room temperature after, reactor assembly is taken out, in culturing room, install, and load onto coil 26;
Then, magnetic valve 10 cuts off the power supply simultaneously with magnetic valve 11, during last liquid, magnetic valve 10 is a closing condition, and magnetic valve 11 is an open mode, open gas meter 17b, nutrient solution is pressed in the reactor body 1 by container for storing liquid, opens inoculation mouthful 4 sealing caps, under gnotobasis, vegetable cell (Saussurea medusa) added on each layer sieve plate in the reactor body 1, build sealing cap, vegetable cell is immersed in the liquid nutrient medium; Simultaneously, coil 26 energisings several minutes, pulling pull bar, make each layer sieve plate from the horizontal by certain angle, open gas meter 17a again and feed sterile air, vegetable cell carries out the full submergence circulation suspension culture of nutrient solution in reactor body 1, and its incubation time sees Table 1; At this moment, vegetable cell in reactor body 1, carry out nutrient solution the full submergence circulation suspension culture phase the cell aggregation flow state as shown in Figure 2.
After full submergence circulation was cultivated, with coil 26 outages, each layer sieve plate fell into horizontal attitude; First magnetic valve 10 and second magnetic valve 16 are switched on simultaneously, this moment, first magnetic valve 10 was an open mode, second magnetic valve 16 is a closing condition, nutrient solution falls back to the container for storing liquid 11 from reactor body 1, vegetable cell drops on each layer sieve plate, carry out the solid-state cultivation of non-submergence of nutrient solution, its incubation time sees Table 1;
With the solid-state cultivation of non-submergence of the full submergence circulation suspension culture of above-mentioned nutrient solution and the nutrient solution several times that hocket, finish until cultivation;
First magnetic valve 10 and second magnetic valve 16 are switched on simultaneously, are cut off the power supply, and realize the loop cycle of nutrient solution between reactor body and container for storing liquid, have simplified the reactor assembly of 4 magnetic valves of original employing.The switch of electromagnetic draw bar device 28, first magnetic valve 10 and second magnetic valve 16 all can be controlled automatically by time controller 23.
Present embodiment also can be looped around a thermostat(t)ed water folder 14 on the container for storing liquid, control culture-liquid temps by thermostat(t)ed water chuck 14 in the culturing process, the nutrient solution that the interval certain hour takes a morsel from container for storing liquid thief hole 21, measure pH, remaining sugar concentration, carbon source, nitrogenous source consumption, the various nutritive salt ionic concn of nutrient solution, so that the cultivation situation in the reactor body 1 is monitored.
In the present embodiment, each layer sieve plate is fixed on the sieve plate bracing frame that stainless steel coil makes the back rotational fixation on the rotation axis on the sieve plate frame by 10 purpose stainless (steel) wires, reach between sieve plate and reactor head and bottom all at regular intervals between each layer sieve plate, 10 purpose stainless (steel) wires can guarantee freely passing through of the interior bubble of nutrient solution, and culture apparatus has the good mixing mass transfer characteristic.The pull bar wheelwork that the wheelwork that sieve plate is swayed is not limited only to the present invention relates to also has many devices that sieve plate is turned an angle, and this belongs to the those skilled in the art required technical knowledge and skills.
Table 1 is adopted culture parameters by present embodiment
| The sieve plate rotational angle | Full submergence circulation incubation time (hour) | The solid-state incubation time of non-submergence (minute) | Light application time (hour/every day) | Intensity of illumination Lux |
| ????60° | ????1 | ????10 | ????16 | ????2500 |
| ????45° | ????2 | ????10 | ????8 | ????4000 |
| ????75° | ????4 | ????10 | ????24 | ????1000 |
| ????50° | ????8 | ????15 | ????10 | ????3500 |
| ????70° | ????2 | ????30 | ????20 | ????1200 |
| ????75° | ????4 | ????40 | ????18 | ????1500 |
| ????45° | ????5 | ????60 | ????15 | ????1400 |
Above-mentioned experimental result shows, is 8 hours at full submergence circulation incubation time, when the solid-state incubation time of non-submergence is 15 minutes, the cell aggregation size to fit, cellular biomass up to 10.4 the gram/every liter (dry weights), the flavones content of cell can reach 4.8%, yield of flavone can reach 0.5 the gram/every liter.
Embodiment 3:
The multilayer for preparing a 50L is rotated the cycle submergence air lift type culture plant cell reactor of sieve plate automatically, as shown in Figure 1, coaxial vertically reactor body 1 center that is fixed on of sieve plate frame that will have 5 layers of stainless steel sieve plate, a steam inlet 24 is set up in container for storing liquid 11 sidewalls bottom, and other 2L structure of reactor with embodiment 1 is identical.
When rotating the cycle submergence air lift type culture plant cell reactor culturing plants cell of sieve plate automatically with embodiment 2 described 50L multilayers, its cultural method is substantially the same manner as Example 1.But in the reactor assembly sterilization process, 121 ℃ of steam that vapour generator is produced enter from the container for storing liquid steam inlet of setting up 24, discharge from the ventage 5 of reactor body 1 and carry out in-situ sterilization.Promptly open gas meter 17b after the nutrient solution sterilization; nutrient solution is pressed into reactor body 1 by container for storing liquid 11; feeding a small amount of sterile air through gas meter 17a protects; treat to open inoculation mouthful 4 sealing caps after culture-liquid temp is reduced to room temperature; under gnotobasis, the culture seed is added; build sealing cap, feed sterile air and cultivate.The nutrient solution that the interval certain hour takes a morsel from container for storing liquid thief hole 21 in the culturing process, the various physical and chemical parameters of mensuration nutrient solution are monitored the cultivation situation in the reactor body.
Its step is as follows:
Behind the in-situ sterilization, nutrient solution in the container for storing liquid is pressed in the reactor body, the pulling pull bar drives the sieve plate that is fixed on the pull bar makes sieve plate become 75 ° of angles with horizontal direction, plant cell cultures on the sieve plate is immersed in the nutrient solution, under the effect of the sterile air that gas distributor 3 feeds, circulate submerged culture 1 or 8 hour;
Spur pull bar again and make sieve plate be returned to level attitude, again with under the nutrient solution in the reactor in container for storing liquid, after plant cell cultures is fallen on the sieve plate, non-submerged culture 10 or 60 minutes;
Repeat submerged culture and non-submerged culturely, finish up to training.
In the present embodiment, as shown in Figure 3, electromagnetic draw bar 27 places in the coil 26, and after the energising, coil 26 control electromagnetic draw bars 27 move up and down, and drives the pull bar 6 control sieve plates that are attached thereto and rotates; Coil 26 is also connected on the time controller 23, and by the automatic rotation of time controller 23 control sieve plates, coil 26 is charged, and under the effect of electromagnetic field, electromagnetic draw bar 27 segment distance that moves upward drives sieve plates by the pull bar in the reactor body 16 and rotates; When the time controller cuts off the power supply, coil 26 outages, magnetic field disappears, and electromagnetic draw bar 27 falls to being returned to state of nature, and the sieve plate in the reactor body 1 also falls back to level attitude thereupon.At electromagnetic draw bar device top one vapor pipe is set, with heat radiation that helps the electromagnetic draw bar main body and the quick elimination of cutting off the power supply the back transient magnetic field, what guarantee sieve plate can fall back to level attitude fast.
Another kind as culture plant cell method of the present invention improves, and by an external light source culture vegetable cell is carried out illumination, and light application time is 8-24 hour, and intensity of illumination is 1000-4000Lux; Also external light source 22 can be connected on the time controller 23, by time controller 23 control light application times.
Table 2 is adopted culture parameters by present embodiment
| The sieve plate rotational angle | Full submergence circulation incubation time (hour) | The solid-state incubation time of non-submergence (minute) | Light application time (hour/every day) | Intensity of illumination Lux |
| ????50° | ????1 | ????10 | ????16 | ????2500 |
| ????65° | ????2 | ????10 | ????8 | ????4000 |
| ????70° | ????4 | ????10 | ????24 | ????1000 |
| ????60° | ????8 | ????10 | ????16 | ????200 |
| ????45° | ????2 | ????60° | ????8 | ????3800 |
| ????75° | ????2 | ????45° | ????24 | ????2000 |
| ????66° | ????4 | ????75° | ????16 | ????3050 |
| ????70° | ????8 | ????60° | ????8 | ????4000 |
The result of comparing embodiment 2 as can be known, along with the increase of reactor volume, the disadvantageous effect of pair cell reduced when sieve plate rotated, the effect of cultivation also can further improve.
In a word, the culture plant cell method and the reactor thereof of cycle submergence air lift type of the present invention, can realize that vegetable cell carries out periodicity submergence, ventilation, periodically illumination, particularly for some vegetable cell that need control the cell aggregation size and need immobilization and suspension culture to combine and cultivate, adopt method and apparatus provided by the present invention, help the transmission that intercellular bioinformation transmission and nutritive substance comprise oxygen and carbon dioxide more, help the growth and the metabolism of cell, have good mixing, mass transfer, heat transfer characteristic.
Claims (10)
1, a kind of cycle submergence air lift type cultural method of vegetable cell, its step is as follows:
1) vegetable cell is inserted on the rotatable sieve plate of multilayer in the transparent reactor main body after the sterilization;
2) plant cell cycle is alternately carried out nutrient solution full buried circulation suspension culture and the solid-state cultivation of the non-buried of nutrient solution, finish until cultivation;
The full buried circulation of described nutrient solution suspension culture is: be under the full buried state of nutrient solution at vegetable cell, rotating rotatable sieve plate makes it horizontal by 45 ° of-75 ° of angles, at this moment, vegetable cell on the sieve plate is suspended in the nutrient solution, carries out the full buried circulation of nutrient solution suspension culture 1-8 hour under the blowing from reactor body bottom gas distributor gas;
The solid-state cultivation of the non-buried of described nutrient solution is: sieve plate is rotated and when being fixed to level attitude, nutrient solution is put into the container for storing liquid that is arranged in its below from the bottom of reactor body, at this moment, vegetable cell drops to and carries out the solid-state cultivation of the non-buried of nutrient solution 10-60 minute on the rotatable sieve plate that is horizontal attitude.
2, press the cycle submergence air lift type cultural method of the described vegetable cell of claim 1, it is characterized in that, the full buried circulation of the nutrient solution that described periodicity hockets suspends and the solid-state cultivation of the non-buried of nutrient solution is carried out under the illumination of the external light source that is positioned at incubator main body periphery, its intensity of illumination is 1000-4000Lux, and light application time is 8-24 hour every day.
3, a kind of cycle submergence air lift type culture apparatus of vegetable cell comprises:
The reactor body that one transparent material is made, fluid infusion mouth on it is connected with a fluid infusion jar by a valve, the reactor body inner bottom part is installed a gas distributor that communicates with source of the gas, the below of reactor body is provided with a container for storing liquid that communicates with the reactor body bottom, this container for storing liquid communicates with source of the gas, it is characterized in that, also comprise be installed in the reactor body can be at the sieve plate that in 45 ° of-75 ° of angular regions, rotates.
4, press the cycle submergence air lift type culture apparatus of the described vegetable cell of claim 3, it is characterized in that, one sieve plate bracing frame is installed in the reactor body, and described sieve plate is fixed on the sieve plate bracing frame by back shaft, and sieve plate is rotating in 45 ° of-75 ° of angular regions around back shaft.
5, press the cycle submergence air lift type culture apparatus of the described vegetable cell of claim 4, it is characterized in that, one end passes the pull bar of reactor body top cover and the outward flange of described sieve plate is fixed, and stretches out the outer tie rod end of reactor body and links to each other with the handle that pull bar is moved up and down.
6, press the cycle submergence air lift type culture apparatus of the described vegetable cell of claim 4, it is characterized in that, one end passes the pull bar of reactor body top cover and the outward flange of described sieve plate is fixed, stretching out the outer tie rod end of reactor body links to each other with the electromagnetic draw bar device that pull bar is moved up and down, described electromagnetic draw bar device is sealed to form after placing an electromagnetic draw bar in the coil, the end and the electromagnetic draw bar of described pull bar are fixed together, and described coil links to each other with a time controller.
7, by the described cycle submergence of claim 6 air lift type culture plant cell reactor, it is characterized in that the top of described electromagnetic draw bar device is provided with vapor pipe.
8, by the described cycle submergence of claim 3 air lift type culture plant cell reactor, it is characterized in that, one magnetic valve that is electrically connected with the time controller is installed on the connecting pipe between described container for storing liquid and the source of the gas, one magnetic valve that is electrically connected with the time controller is installed on the connecting pipe of container for storing liquid inlet mouth and reactor body air, two magnetic valves open/close opposite states.
By the described cycle submergence of claim 3 air lift type culture plant cell reactor, it is characterized in that 9, be evenly distributed with external light source around the described reactor body, its intensity of illumination is 1000-4000Lux.
10, by the described cycle submergence of claim 3 air lift type culture plant cell reactor, it is characterized in that suit one thermostat(t)ed water chuck around the described container for storing liquid.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101786765A (en) * | 2010-02-11 | 2010-07-28 | 宁波新芝生物科技股份有限公司 | Batch immersion bioreactor |
| CN101518219B (en) * | 2009-04-03 | 2011-06-15 | 西北工业大学 | Apparatus for magnetic suspension culture of aquatic organisms |
| WO2012061950A1 (en) * | 2010-11-09 | 2012-05-18 | Universidad De Santiago De Chile | Thermo-photo-bioreactor and method for the culture and mass micropropagation of deschampsia antarctica in vitro |
| CN111670811A (en) * | 2020-07-31 | 2020-09-18 | 广西壮族自治区农业科学院 | Constant temperature device for cell tissue culture |
| CN113337399A (en) * | 2021-04-26 | 2021-09-03 | 张庆兰 | Plant cell culture device of daylighting floats |
| US11299700B1 (en) | 2021-02-19 | 2022-04-12 | Acequia Biotechnology, Llc | Bioreactor containers and methods of growing hairy roots using the same |
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2004
- 2004-01-20 CN CN 200410000797 patent/CN1648240A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101518219B (en) * | 2009-04-03 | 2011-06-15 | 西北工业大学 | Apparatus for magnetic suspension culture of aquatic organisms |
| CN101786765A (en) * | 2010-02-11 | 2010-07-28 | 宁波新芝生物科技股份有限公司 | Batch immersion bioreactor |
| CN101786765B (en) * | 2010-02-11 | 2012-01-04 | 宁波新芝生物科技股份有限公司 | Batch immersion bioreactor |
| WO2012061950A1 (en) * | 2010-11-09 | 2012-05-18 | Universidad De Santiago De Chile | Thermo-photo-bioreactor and method for the culture and mass micropropagation of deschampsia antarctica in vitro |
| CN111670811A (en) * | 2020-07-31 | 2020-09-18 | 广西壮族自治区农业科学院 | Constant temperature device for cell tissue culture |
| US11299700B1 (en) | 2021-02-19 | 2022-04-12 | Acequia Biotechnology, Llc | Bioreactor containers and methods of growing hairy roots using the same |
| CN113337399A (en) * | 2021-04-26 | 2021-09-03 | 张庆兰 | Plant cell culture device of daylighting floats |
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