CN102746987A - Peristaltic pump-type intermittent immersion tissue culture device - Google Patents
Peristaltic pump-type intermittent immersion tissue culture device Download PDFInfo
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- CN102746987A CN102746987A CN2012102138591A CN201210213859A CN102746987A CN 102746987 A CN102746987 A CN 102746987A CN 2012102138591 A CN2012102138591 A CN 2012102138591A CN 201210213859 A CN201210213859 A CN 201210213859A CN 102746987 A CN102746987 A CN 102746987A
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- peristaltic pump
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- silica gel
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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
- C12M23/40—Manifolds; Distribution pieces
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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
- 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
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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
- C12M23/58—Reaction vessels connected in series or in parallel
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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
- C12M29/12—Pulsatile flow
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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
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/02—Filters
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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Abstract
The invention relates to a peristaltic pump-type intermittent immersion tissue culture device. The peristaltic pump-type intermittent immersion tissue culture device comprises culture containers, a liquid separator, a nutrient solution storage container and an actuating and regulation mechanism. The culture containers are multiple lower-opening bottles with covers and are arranged in parallel. Each adjacent two of the culture containers are communicated by an aseptic silicone tube arranged on the covers of the culture containers. The liquid separator is provided with branch interfaces and a total interface. The total interface is connected to the nutrient solution storage container by an aseptic silicone tube. The branch interfaces are connected to lower openings of the culture containers by aseptic silicone tubes. The aseptic silicone tube connected to the nutrient solution storage container stretches to the bottom of the nutrient solution storage container. The actuating and regulation mechanism is arranged on the aseptic silicone tube located between the nutrient solution storage container and the liquid separator. The peristaltic pump-type intermittent immersion tissue culture device has the advantages that a structure is simple; a manufacturing cost is low; use is convenient; motive power consumption and noise of a peristaltic pump are low; nutrient solution transmission is completely enclosed; in work, pollution probability is small; an occupied space is small; a culture space is saved; and a tissue culture cost is reduced.
Description
Technical field
The present invention relates to a kind of plant tissue culture device, the power wheel drive of specifically a kind of employing peristaltic pump reaches the intermittently tissue culture device of submerged culture explant purpose, belongs to biological technical field.
Background technology
Plant tissue culture technique is from being born so far; The detoxification and the fast numerous aspect that are widely used in crop breeding, flowers, fruit tree, vegetables, forest and herbal medicine; Have the incomparable advantage of other technologies means, become one of indispensable technology of modern biotechnology.
At present, adopt the plant tissue culture technique of liquid culture in large-scale industrialized production, to replace traditional solid, semi-solid training method gradually.Liquid culture has and reduces cost, and reduces the demand to the labor force, raises the efficiency; Be convenient to the characteristics of scale operation, the intermittent immersed liquid culture that especially near more than ten years grow up has possessed the semi-solid common advantage of cultivating of liquid culture and solid; Have following advantage: 1. intermittently submergence, gas replenishment is abundant, and oxygen is sufficient; Reduced the probability that is prone to produce lopsided seedling and height vitrifying seedling during common liq is cultivated, the quality of cultivating seedling is improved dry weight; Weight in wet base, breeding potential all obviously improve, and the domestication surviving rate also increases substantially; 2. the robotization control that has realized nutritive medium down assisting of computer program is supplied and is drained, and has increased technology content, has reduced work density, has practiced thrift a large amount of labor forces, has reduced cost.The representational achievement in research of intermittent immersed culture apparatus is an intermittent type solution-air exchange bio-reactor; Air lift type (RITA
(WO96/25484)) and double cylinder (BIT
) bio-reactor are arranged, and both common features are: the air pressure that 1. utilizes the air lift pump generation is as power; 2. can keep good ventilated; 3. can cultured continuously reduce subculture number; 4. be easy to various culture condition; 5. regeneration plant resistance significantly strengthens.Gas-powered liquid-flow energy consumption is high, noise is big but they also exist simultaneously; Gas-powered is unstable; Nutrient solution upgrades complicated operation; And RITA
(the Recipient for Automated Temporary Immersion system; Robotization immersion system culture vessel at intermittence) complex structure makes manufacturing cost too high; BIT
(the Twin flasks system; The double cylinder system) culturing bottle needs too much, and the unit space utilization ratio is reduced, and has limited their widespread again.
Summary of the invention
Be to solve the deficiency of prior art, the object of the present invention is to provide intermittently submergence tissue culture device of a kind of simple in structure, peristaltic pump formula that cost is low, easy to use.
For achieving the above object, the present invention realizes through following technical scheme:
A kind of peristaltic pump formula is the submergence tissue culture device intermittently, it is characterized in that, comprises culture vessel, skimmer, nutritive medium liquid storage container and power and regulating and controlling mechanism; Described culture vessel is a tubualted bottle with cover; And be set side by side with many groups, then be communicated with through the aseptic silica gel pipe that is arranged on the culture vessel lid between the adjacent culture vessel, described skimmer is provided with branch interface and total interface; Described total interface is connected with the nutritive medium liquid storage container through the aseptic silica gel pipe; Interface then was connected through the end opening of aseptic silica gel pipe with many group culture vessels in described minute, and wherein, the aseptic silica gel pipe that is connected with the nutritive medium liquid storage container extend into the bottle end of nutritive medium liquid storage container; In addition, described power and regulating and controlling mechanism then are arranged on the aseptic silica gel pipe between nutritive medium liquid storage container and the skimmer.
Further; Described power and regulating and controlling mechanism comprise peristaltic pump and time-controlling arrangement; Described time-controlling arrangement and peristaltic pump are integrated in the controlling box; And be interconnected, and described peristaltic pump pump head stretches out controlling box, and the aseptic silica gel tube portion between described nutritive medium liquid storage container and the skimmer is arranged in the groove of peristaltic pump pump head.
And described time-controlling arrangement comprises timer and rly..
In addition, described nutritive medium liquid storage container is a lidded container, and the container cover of nutritive medium liquid storage container has first sterilizing filter through the aseptic silica gel pipe coupling.
And be provided with second sterilizing filter on the aseptic silica gel pipe at the end opening place of described many group culture vessels.
And through the aseptic silica gel pipe coupling the 3rd sterilizing filter is arranged foremost with on the lid of the culture vessel of rearmost end in the culture vessel that described many groups are set up in parallel.
The invention has the beneficial effects as follows: (1) the present invention is simple in structure, and low cost of manufacture is easy to use; (2) the present invention has saved energy consumption, has reduced noise through changing traditional gas-powered mode into the peristaltic pump type of drive; (3) nutritive medium transmission complete closed is aseptic, and the probability that pollutes during work is little; (4) the present invention can will be installed on the various culturing racks according to actual, and it is little to take up room, and has practiced thrift culture space, has reduced group training cost.
Description of drawings
Fig. 1 is the structural representation of one embodiment of the invention.
The main mark implication is following among the figure:
1, culture vessel 2, skimmer 3, nutritive medium liquid storage container 4, peristaltic pump
5, controlling box 6, timer 7, rly. 8, power supply
9, first sterilizing filter 10, second sterilizing filter 11, the 3rd sterilizing filter.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is done concrete introduction.
Fig. 1 is the structural representation of one embodiment of the invention.
As shown in Figure 1: a kind of peristaltic pump formula is the submergence tissue culture device intermittently; Comprise culture vessel 1, skimmer 2, nutritive medium liquid storage container 3 and power and regulating and controlling mechanism, work that described culture vessel 1 is a tubualted bottle with cover but described culture vessel 1, skimmer 2, nutritive medium liquid storage container 3 and power and regulating and controlling mechanism level are installed on the common culturing rack; And be set side by side with many groups; In this embodiment, described culture vessel 1 is three groups, then is communicated with through the aseptic silica gel pipe that is arranged on the culture vessel lid between the adjacent culture vessel 1; Realize that ventilation is unimpeded, guarantee that each culture vessel 1 internal gas pressure is consistent.Described skimmer 2 is provided with branch interface and total interface; Described total interface is connected with nutritive medium liquid storage container 3 through the aseptic silica gel pipe; Interface then was connected with the end opening of three groups of culture vessels 1 respectively through the aseptic silica gel pipe in described minute, and wherein, the aseptic silica gel pipe that is connected with nutritive medium liquid storage container 3 extend into the bottle end of nutritive medium liquid storage container 3; In addition; Described power and regulating and controlling mechanism then are arranged on the aseptic silica gel pipe between nutritive medium liquid storage container 3 and the skimmer 2, and in this embodiment, described time-controlling arrangement comprises timer 6 and rly. 7.
Be specially: described power and regulating and controlling mechanism comprise peristaltic pump 4 and time-controlling arrangement; Wherein, described time-controlling arrangement is used to set the time of each process step of peristaltic pump 4 one actions circulation, and described time-controlling arrangement and peristaltic pump 4 are integrated in the controlling box 5; And time-controlling arrangement and peristaltic pump 4 are interconnected; And described peristaltic pump pump head stretches out controlling box 5, and the aseptic silica gel tube portion between described nutritive medium liquid storage container 3 and the skimmer 2 is arranged in the groove of peristaltic pump pump head, and described peristaltic pump 4 is all supplied power through power supply 8 with time-controlling arrangement; The peristaltic pump pump head is under the motor forward or reverse drives; Positive dirction or continue in the other direction extruding aseptic silica gel pipe produces positive dirction or reciprocal pressure, makes nutritive medium in the nutritive medium storage vessel 3 by sucking-off or suction.
In this embodiment, described nutritive medium liquid storage container 3 is a lidded container, and the container cover of nutritive medium liquid storage container 3 has first sterilizing filter 9 through the aseptic silica gel pipe coupling, and this interface can be used for replenishing and changing nutritive medium.And be provided with second sterilizing filter 10 on the aseptic silica gel pipe at the end opening place of described three groups of culture vessels 1.And through the aseptic silica gel pipe coupling the 3rd sterilizing filter 11 is arranged foremost with on the lid of the culture vessel of rearmost end in described three groups of culture vessels that are set up in parallel 1; Above-mentioned institute's first bacterium strainer and the 3rd sterilizing filter are used for filtrated air; Make a bottle interior air cleaning; The second above-mentioned sterilizing filter is used to filter nutritive medium, avoids polluting.
In this embodiment, the aseptic silica gel pipe between described culture vessel 1, skimmer 2 and the nutritive medium liquid storage container 3 has constituted the passage of nutritive medium transmission, and makes and form closed system between culture vessel 1, skimmer 2 and the nutritive medium liquid storage container 3.
Working process of the present invention is: earlier time-controlling arrangement is set " just changeing the time, just changeing rest time after back rest time, reversing time, the counter-rotating ", peristaltic pump 4 promptly carries out work according to the time of setting.At first; Peristaltic pump 4 forward rotation; The aseptic silica gel pipe that the pump head extruding is in contact with it, the nutritive medium in the nutritive medium storage vessel 3 is drawn out of, and flows into each culture vessel 1 through skimmer 2; Nutritive medium reaches the certain liquid height in culture vessel 1 after (whole or most of submerged culture things usually), peristaltic pump 4 quits work to get into is just changeing the back stationary state.Nutrition this moment liquid level is stabilized in the submergence height, and culture is in submerged culture state, fully carries out the absorption of nutrient ingredients exchange.After accomplishing rest time, peristaltic pump 4 starts and beginning reverse rotation are slowly extracted nutritive medium out the nutritive medium storage vessel that also refluxes again until draining from culture vessel 1, and peristaltic pump 4 quits work and gets into counter-rotating back stationary state.1 li no nutritive medium of culture vessel this moment, culture can fully carry out gaseous interchange, realizes intermittently submerged culture purpose.Through " static after just changeing → just changeing → counter-rotating → counter-rotating back is static " four steps of above-mentioned peristaltic pump 4, accomplish once intermittently submerged culture circulation, different cultures submerged culture net cycle time at required intermittence can specifically be set according to actual needs.
The foregoing description does not limit the present invention in any form, and all employings are equal to the technical scheme that mode obtained of replacement or equivalent transformation, all drop in protection scope of the present invention.
Claims (6)
1. peristaltic pump formula submergence at an intermittence tissue culture device is characterized in that, comprises culture vessel, skimmer, nutritive medium liquid storage container and power and regulating and controlling mechanism; Described culture vessel is a tubualted bottle with cover; And be set side by side with many groups, then be communicated with through the aseptic silica gel pipe that is arranged on the culture vessel lid between the adjacent culture vessel, described skimmer is provided with branch interface and total interface; Described total interface is connected with the nutritive medium liquid storage container through the aseptic silica gel pipe; Interface then was connected through the end opening of aseptic silica gel pipe with many group culture vessels in described minute, and wherein, the aseptic silica gel pipe that is connected with the nutritive medium liquid storage container extend into the bottle end of nutritive medium liquid storage container; In addition, described power and regulating and controlling mechanism then are arranged on the aseptic silica gel pipe between nutritive medium liquid storage container and the skimmer.
2. peristaltic pump formula according to claim 1 is the submergence tissue culture device intermittently; It is characterized in that; Described power and regulating and controlling mechanism comprise peristaltic pump and time-controlling arrangement, and described time-controlling arrangement and peristaltic pump are integrated in the controlling box, and are interconnected; And described peristaltic pump pump head stretches out controlling box, and the aseptic silica gel tube portion between described nutritive medium liquid storage container and the skimmer is arranged in the groove of peristaltic pump pump head.
3. peristaltic pump formula according to claim 2 is the submergence tissue culture device intermittently, it is characterized in that described time-controlling arrangement comprises timer and rly..
4. peristaltic pump formula according to claim 1 is the submergence tissue culture device intermittently, it is characterized in that described nutritive medium liquid storage container is a lidded container, and the container cover of nutritive medium liquid storage container has first sterilizing filter through the aseptic silica gel pipe coupling.
5. peristaltic pump formula according to claim 1 is the submergence tissue culture device intermittently, it is characterized in that, is provided with second sterilizing filter on the aseptic silica gel pipe at the end opening place of described many group culture vessels.
6. peristaltic pump formula according to claim 1 is the submergence tissue culture device intermittently, it is characterized in that, through the aseptic silica gel pipe coupling the 3rd sterilizing filter is arranged foremost with on the lid of the culture vessel of rearmost end in the culture vessel that described many groups are set up in parallel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102138591A CN102746987A (en) | 2012-06-27 | 2012-06-27 | Peristaltic pump-type intermittent immersion tissue culture device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102138591A CN102746987A (en) | 2012-06-27 | 2012-06-27 | Peristaltic pump-type intermittent immersion tissue culture device |
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| CN102746987A true CN102746987A (en) | 2012-10-24 |
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| CN2012102138591A Pending CN102746987A (en) | 2012-06-27 | 2012-06-27 | Peristaltic pump-type intermittent immersion tissue culture device |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191798A (en) * | 2015-09-29 | 2015-12-30 | 中国科学院植物研究所 | Temporary-immersion lily scale rapid-propagation method |
| CN105210867A (en) * | 2015-09-29 | 2016-01-06 | 中国科学院植物研究所 | A kind of propagation quick-breeding method of intermittent immersed pteridophyte green spheroids |
| CN105210866A (en) * | 2015-09-29 | 2016-01-06 | 中国科学院植物研究所 | A kind of intermittent immersed orchid protocorms propagation quick-breeding method |
| CN105340733A (en) * | 2015-09-29 | 2016-02-24 | 中国科学院植物研究所 | Intermittent immersion type gesneriaceae leave adventitious buds induction method |
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| CN1743450A (en) * | 2005-09-15 | 2006-03-08 | 上海交通大学 | Scierotomal cell amplifying bioreactor system |
| CN102174397A (en) * | 2011-03-07 | 2011-09-07 | 四川大学 | Bionic three-dimensional fluid shear stress cell culture device and shear stress loading method thereof |
| CN102286374A (en) * | 2011-07-26 | 2011-12-21 | 江苏农林职业技术学院 | Plant tissue liquid automatic intermittent submerged circulative culture device |
| CN202643701U (en) * | 2012-06-27 | 2013-01-02 | 江苏农林职业技术学院 | Peristaltic pump type intermittent immersion tissue culture device |
-
2012
- 2012-06-27 CN CN2012102138591A patent/CN102746987A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1743450A (en) * | 2005-09-15 | 2006-03-08 | 上海交通大学 | Scierotomal cell amplifying bioreactor system |
| CN102174397A (en) * | 2011-03-07 | 2011-09-07 | 四川大学 | Bionic three-dimensional fluid shear stress cell culture device and shear stress loading method thereof |
| CN102286374A (en) * | 2011-07-26 | 2011-12-21 | 江苏农林职业技术学院 | Plant tissue liquid automatic intermittent submerged circulative culture device |
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Non-Patent Citations (1)
| Title |
|---|
| H. ETIENNE 等: "Temporary immersion systems in plant micropropagation", 《PLANT CELL, TISSUE AND ORGAN CULTURE》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191798A (en) * | 2015-09-29 | 2015-12-30 | 中国科学院植物研究所 | Temporary-immersion lily scale rapid-propagation method |
| CN105210867A (en) * | 2015-09-29 | 2016-01-06 | 中国科学院植物研究所 | A kind of propagation quick-breeding method of intermittent immersed pteridophyte green spheroids |
| CN105210866A (en) * | 2015-09-29 | 2016-01-06 | 中国科学院植物研究所 | A kind of intermittent immersed orchid protocorms propagation quick-breeding method |
| CN105340733A (en) * | 2015-09-29 | 2016-02-24 | 中国科学院植物研究所 | Intermittent immersion type gesneriaceae leave adventitious buds induction method |
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Application publication date: 20121024 |