CN205501326U - Centrifugal sedimentation formula bioreactor for plant cell continuous culture - Google Patents
Centrifugal sedimentation formula bioreactor for plant cell continuous culture Download PDFInfo
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- CN205501326U CN205501326U CN201620209653.5U CN201620209653U CN205501326U CN 205501326 U CN205501326 U CN 205501326U CN 201620209653 U CN201620209653 U CN 201620209653U CN 205501326 U CN205501326 U CN 205501326U
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Abstract
The utility model discloses a centrifugal sedimentation formula bioreactor for plant cell continuous culture, including the whole main part that is U type structure, it rises liquid portion one side and still is equipped with the inlet, and the bottom position that falls liquid portion is equipped with the cell harvesting mouth, it still is equipped with the gas blowout head to rise liquid portion on the bottom position that lies in the main part, fall liquid portion from last to having sealing funnel and spiral pipe, the sealing funnel top is sealed with the inner wall laminating of the liquid portion of falling of main part, and the bottom of sealing funnel is connected in the entrance of spiral pipe, and the bottom of spiral pipe department still connects gravity sedimentation pipe, and gravity sedimentation pipe one side in addition connects waste liquid collector. The utility model discloses an in the bioreactor with the spiral pipe at the bottom of will realize the separation of cell and culture solution in the gravity sedimentation pipe that links to each other, can obviously improve the efficiency that the cell was held back after appending to this spiral pipe on the reactor to realize the continuous perfusion culture of plant cell, and then improve the efficiency that plant cell cultivateed.
Description
Technical field
This utility model relates to culture plant cell method and cultivates the bioreactor technology field adapted to, and particularly relates to centrifugal type centrifugal type plant cell cultivation bioreactor continuously.
Background technology
The focus of the exploitation of phytochemicals production always people's research, utilizes bioreactor to carry out Vitro Plant tissue culture and is possible not only to produce many secondary metabolites with important value, and can alleviate excessively excavating of wild resource.But, the middle secondary metabolite that plant cell produces in incubation accumulation in culture fluid can suppress the further synthesis of product.Cell can be discharged into the material inside culture medium and constantly take to outside reactor by plant cell continuous perfusion culture, efficiently solves cell brown stain and the problem of product feedback suppression.Compared with batch culture, plant cell is cultivated continuously the biggest advantage, but, during plant cell continuous perfusion culture, the separation efficiency of cell and culture fluid becomes and limits the principal element that this technology develops further.It is currently used for the anti-device type that plant cell cultivates continuously mainly to have: film filtering reactor and gravitational settling reactor etc..Membrane filtration easily blocks, so being mainly used in zooblast；Gravitational settling efficiency is the lowest can not large-scale promotion application.
At present, Authorization Notice No. is CN200710044223.8, the Chinese patent of entitled medicinal plants histiocyte continuous perfusion culture Airlift type bioreactor system, have been disclosed for the medicinal plants histiocyte continuous perfusion culture Airlift type bioreactor system of a kind of biological technical field, comprising: the continuous harvesting apparatus of liquid containing bottle, reactor body, gravity settling device, waste liquid bottle, tissue/organ, duplex sampling bottle, liquid pump, oxygen probe, PH probe, on-line monitoring system, computer.Liquid containing bottle, liquid pump, reactor body, gravity sink device, waste liquid bottle are sequentially connected by silica gel tube.Oxygen probe, PH probe is connected with computer by on-line monitoring system.This utility model has reactor assembly and can be used for the cultivation of various different tissues, cell and organ, has saved cost, avoids the advantages such as the damage that causes plant tissue cell, is applied to medicinal plants tissue, cell and organ suspension culture.
But the plant cell intercepting and capturing rate of above-mentioned bioreactor is not the highest, can cause certain waste, owing to plant cell itself obtains the most relatively difficult, cost is the most higher.
Utility model content
In order to overcome above-mentioned the deficiencies in the prior art, this utility model provides, centrifugal type plant cell is cultivated with bioreactor continuously, its object is to use outer circulation type reactor, decline district at it and be provided with a serpentine pipe and gravitational settling pipe, along with culture fluid circulates at inside reactor, cell is centrifuged formula in serpentine pipe and rotates and be deposited to bottom serpentine pipe, so separating with culture fluid with realizing cell in the gravitational settling pipe being connected at the bottom of serpentine pipe.The efficiency of cell retention can be significantly improved after being attached on reactor by this serpentine pipe, thus realize plant cell continuous perfusion culture, and then improve the efficiency of culture plant cell, shorten its cultivation cycle.
This utility model the technical scheme is that centrifugal type plant cell cultivation bioreactor continuously, main body including overall U-shaped structure, main body includes liter liquid portion and descending liquid portion, rises liquid portion and the bottom of its correspondence of descending liquid portion and tip position UNICOM respectively；Rising side, liquid portion and be additionally provided with inlet, the bottom position in descending liquid portion is provided with cell harvesting mouth；Rise liquid portion on the bottom position be positioned at main body, be additionally provided with gas blowout head, gas blowout head ejection air-flow upwards, and then drive the culture fluid risen in liquid portion to rise；Descending liquid portion is provided with the inwall gluing, sealing in the descending liquid portion of sealing funnel and serpentine pipe, sealing funnel top and main body from top to bottom, and the bottom of sealing funnel is connected to the porch of serpentine pipe.Bioreactor of the present utility model uses outer circulation type reactor, decline district at it and be provided with a serpentine pipe, along with culture fluid circulates at inside reactor, cell is centrifuged formula in serpentine pipe and rotates and be deposited to bottom serpentine pipe, so serpentine pipe will realize cell and separate with culture fluid.The efficiency of cell retention can be significantly improved after being attached on reactor by this serpentine pipe, thus realize plant cell continuous perfusion culture, and then improve the efficiency of culture plant cell, shorten its cultivation cycle.
Further, serpentine pipe is also associated with gravitational settling pipe, and the bottom of serpentine pipe is connected with the bottom of gravitational settling pipe, and the top of gravitational settling pipe is waste liquid outlet, it is ensured that gravitational settling occurs after centrifugal sedimentation.
Further, the diameter of gravitational settling pipe is less than the diameter of serpentine pipe, and the diameter of gravitational settling pipe in proportion can not be less than the 1/3 of the descending liquid portion diameter of main body.
Further, the bottom of serpentine pipe is horn-like outlet, it is simple to cell settlement reactor bottom.
Further, between sealing funnel and descending liquid portion inwall, it is additionally provided with sealing ring, improves the seal degree of sealing funnel and descending liquid portion inwall, it is ensured that culture fluid can be partially into serpentine pipe from leting slip a remark of sealing funnel completely.
Further, the tip position of main body is additionally provided with air filter, crosses the bulky grain thing pollutant in air filtering, prevents from polluting culture fluid, can guarantee that again culture fluid can breathe enough air simultaneously.
Further, the external connection air pump of gas blowout head, it is additionally provided with glass filter element inside its gas blowout head.
Compared with prior art, the beneficial effects of the utility model are: bioreactor of the present utility model uses outer circulation type reactor, decline district at it and be provided with a serpentine pipe, along with culture fluid circulates at inside reactor, cell is centrifuged formula in serpentine pipe and rotates and be deposited to bottom serpentine pipe, so serpentine pipe will realize cell and separate with culture fluid.The efficiency of cell retention can be significantly improved after being attached on reactor by this serpentine pipe, thus realize plant cell continuous perfusion culture, and then improve the efficiency of culture plant cell.
Accompanying drawing explanation
Fig. 1 is the structural representation of an embodiment of centrifugal type plant cell cultivation bioreactor continuously；
Wherein: 1-inlet, 2-rises liquid portion, 3-gas blowout head；4-cell harvesting mouth, 5-serpentine pipe, 51-entrance, bottom 52-；6-sealing funnel, 7-gravitational settling pipe, 8-air filter, 9-main body, 10-descending liquid portion, 11-air pump, 12-culture fluid, 13-waste liquid outlet, 14-sealing ring.
Detailed description of the invention
In order to deepen understanding of the present utility model, further illustrating this utility model with embodiment below in conjunction with the accompanying drawings, this embodiment is only used for explaining this utility model, does not constitute protection domain of the present utility model and limits.
Embodiment 1
As it is shown in figure 1, centrifugal type plant cell is cultivated with bioreactor continuously, including the main body 9 of overall U-shaped structure, main body 9 includes liter liquid portion 2 and descending liquid portion 10, rises liquid portion 2 and the bottom of its correspondence of descending liquid portion 10 and tip position UNICOM respectively；Rising side, liquid portion 2 and be additionally provided with inlet 1, the bottom position in descending liquid portion 10 is provided with cell harvesting mouth 4；Rise liquid portion 2 on the bottom position being positioned at main body 9, be additionally provided with gas blowout 3, gas blowout 3 ejection air-flow upwards, and then drive the culture fluid 12 risen in liquid portion 2 to rise；Descending liquid portion 10 is provided with the inwall gluing, sealing in the descending liquid portion 10 of sealing funnel 6 and serpentine pipe 5, sealing funnel 6 top and main body 9 from top to bottom, and the bottom of sealing funnel 6 is connected at the entrance 51 of serpentine pipe 5.Bioreactor of the present utility model uses outer circulation type reactor, decline district at it and be provided with a serpentine pipe, along with culture fluid circulates at inside reactor, cell is centrifuged formula in serpentine pipe and rotates and be deposited to bottom serpentine pipe, so serpentine pipe will realize cell and separate with culture fluid.The efficiency of cell retention can be significantly improved after being attached on reactor by this serpentine pipe, thus realize plant cell continuous perfusion culture, and then improve the efficiency of culture plant cell, reduce its toxigenic capacity.
In the above-described embodiments, serpentine pipe 5 is also associated with gravitational settling pipe 7, and the bottom 52 of serpentine pipe 5 is connected with the bottom of gravitational settling pipe 7, and the top of gravitational settling pipe 7 is waste liquid outlet 13, it is ensured that gravitational settling occurs after centrifugal sedimentation.The diameter of gravitational settling pipe 7 is less than the diameter of serpentine pipe 5, and the diameter of gravitational settling pipe 7 in proportion can not be less than the 1/3 of descending liquid portion 10 diameter of main body 9.
In the above-described embodiments, the bottom 52 of serpentine pipe 5 is horn-like outlet, it is simple to cell settlement reactor bottom.
In the above-described embodiments, between sealing funnel 6 and descending liquid portion 10 inwall, it is additionally provided with sealing ring 14, improves the seal degree of sealing funnel and descending liquid portion inwall, it is ensured that culture fluid can be partially into serpentine pipe from leting slip a remark of sealing funnel completely.
In the above-described embodiments, the tip position of main body 9 is additionally provided with air filter 8, crosses the bulky grain thing pollutant in air filtering, prevents from polluting culture fluid.
In the above-described embodiments, the external connection air pump 11 of gas blowout 3, its gas blowout 3 is internal is additionally provided with glass filter element.In the above-described embodiments, the outside of gas blowout 3 can be connected with air pump 11, the internal glass filter element that is additionally provided with of gas blowout 3, and the bubble diameter formed when a diameter of 180 μm of glass filter element is little, it is uniformly distributed, damage to plant cell when effectively reducing bubbles burst.
Embodiment 2
Use the method that cell is cultivated by bioreactor continuously, comprise the steps: that the MS culture medium using culture fluid to be interpolation NAA and IAA is cultivated in plant cell is inserted U-shaped centrifugal type bioreactor, keeping in gas blowout head Ventilation Rate out is 200-300vvm, cultivation temperature is 20-30 DEG C, cultivates 25-35 days.
In the above-described embodiments, reactor includes the main body 9 of overall U-shaped structure, and main body 9 includes liter liquid portion 2 and descending liquid portion 10, rises liquid portion 2 and the bottom of its correspondence of descending liquid portion 10 and tip position UNICOM respectively；Rising side, liquid portion 2 and be additionally provided with inlet 1, the bottom position in descending liquid portion 10 is provided with cell harvesting mouth 4；Rise liquid portion 2 on the bottom position being positioned at main body 9, be additionally provided with gas blowout 3, gas blowout 3 ejection air-flow upwards, and then drive the culture fluid 12 risen in liquid portion 2 to rise；Descending liquid portion 10 is provided with sealing funnel 6 and serpentine pipe 5 from top to bottom, the inwall gluing, sealing in the descending liquid portion 10 of sealing funnel 6 top and main body 9, the bottom of sealing funnel 6 is connected at the entrance 51 of serpentine pipe 5, being also connected with gravitational settling pipe 7 at the bottom 52 of serpentine pipe 5, the bottom 52 of serpentine pipe 5 is horn mouth horn shape down.
In the above-described embodiments, the irrigation rate ratio of the perfusion plant cell of inlet 1 is 10%, and irrigation rate accounts for the percentage ratio of volume than the volume for flowing to every day, and its unit is V/V.
In the above-described embodiments, the tip position of main body 9 is additionally provided with air filter 8.
In the above-described embodiments, being to add in plant cell is inserted U-shaped centrifugal type bioreactor by the MS culture medium of NAA and IAA to cultivate with culture fluid, keeping in gas blowout head Ventilation Rate out is 260vvm, and cultivation temperature is 25 DEG C, cultivates 29 days.Bioreactor of the present utility model uses outer circulation type reactor, decline district at it and be provided with a serpentine pipe and gravitational settling pipe, along with culture fluid circulates at inside reactor, cell is centrifuged formula in serpentine pipe and rotates and be deposited to bottom serpentine pipe, so separating with culture fluid with realizing cell in the gravitational settling pipe being connected at the bottom of serpentine pipe.The efficiency of cell retention can be significantly improved after being attached on reactor by this serpentine pipe, thus realize plant cell continuous perfusion culture, and then improve the efficiency of culture plant cell.
Embodiment 3
The device that the present embodiment uses is as shown in Figure 1.By the present embodiment, Glycyrrhiza cell having been carried out continuous cultivation, initial inoculation density is 5g (dry weight)/L, and culture fluid is the MS culture medium adding NAA and IAA, and Ventilation Rate is 300vvm, and cultivation temperature is 25 DEG C.Simultaneously with Glycyrrhiza cell for material this reactor cut efficiency and kLa is determined.Result shows, under conditions of being suitable for plant cell growth, (Ventilation Rate is 0.3vvm, irrigation rate is 10%), cell retention efficiency reaches 100%, volume dissolved oxygen coefficient k La is 5.0, after 27 days, cell maximum biomass reaches 14.0 g/ L, higher than maximum biomass during batch culture 12.0 g/ L.Device involved by this utility model is very suitable for plant cell and cultivates continuously, can be in large-scale application.
What embodiment of the present utility model was announced is preferred embodiment; but it is not limited thereto; those of ordinary skill in the art; easily according to above-described embodiment; understand spirit of the present utility model; and make different amplifications and change, but as long as without departing from spirit of the present utility model, all in protection domain of the present utility model.
Claims (7)
1. centrifugal type plant cell is cultivated with bioreactor continuously, it is characterized in that: include the main body (9) of overall U-shaped structure, described main body (9) includes liter liquid portion (2) and descending liquid portion (10), described liter of liquid portion (2) and the bottom of descending liquid portion (10) its correspondence and tip position UNICOM respectively；Described liter of liquid portion (2) side is additionally provided with inlet (1), and the bottom position of described descending liquid portion (10) is provided with cell harvesting mouth (4)；
Described liter of liquid portion (2) is additionally provided with gas blowout head (3), described gas blowout head (3) ejection air-flow upwards on the bottom position being positioned at main body (9), and then drives the culture fluid (12) risen in liquid portion (2) to rise；
Described descending liquid portion (10) is provided with sealing funnel (6) and serpentine pipe (5) from top to bottom, the inwall gluing, sealing in the descending liquid portion (10) of described sealing funnel (6) top and main body (9), the bottom of described sealing funnel (6) is connected to entrance (51) place of serpentine pipe (5).
Centrifugal type plant cell the most according to claim 1 is cultivated with bioreactor continuously, it is characterized in that: described serpentine pipe (5) is also associated with gravitational settling pipe (7), and the bottom (52) of described serpentine pipe (5) is connected with the bottom of gravitational settling pipe (7), the top of described gravitational settling pipe (7) is waste liquid outlet (13), it is ensured that gravitational settling occurs after centrifugal sedimentation.
Centrifugal type plant cell the most according to claim 2 is cultivated with bioreactor continuously, it is characterized in that: the diameter of described gravitational settling pipe (7) less than the diameter of serpentine pipe (5), and the diameter of described gravitational settling pipe (7) can not be less than in proportion main body (9) descending liquid portion (10) diameter 1/3.
Centrifugal type plant cell the most according to claim 2 is cultivated with bioreactor continuously, it is characterised in that: the bottom (52) of described serpentine pipe (5) is horn-like outlet, it is simple to cell settlement reactor bottom.
Centrifugal type plant cell the most according to claim 1 is cultivated with bioreactor continuously, it is characterised in that: it is additionally provided with sealing ring (14) between described sealing funnel (6) and descending liquid portion (10) inwall.
Centrifugal type plant cell the most according to claim 1 is cultivated with bioreactor continuously, it is characterised in that: the tip position of described main body (9) is additionally provided with air filter (8).
Centrifugal type plant cell the most according to claim 1 is cultivated with bioreactor continuously, it is characterised in that: the external connection air pump (11) of described gas blowout head (3), its gas blowout head (3) is internal is additionally provided with glass filter element.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620209653.5U CN205501326U (en) | 2016-03-18 | 2016-03-18 | Centrifugal sedimentation formula bioreactor for plant cell continuous culture |
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| CN201620209653.5U CN205501326U (en) | 2016-03-18 | 2016-03-18 | Centrifugal sedimentation formula bioreactor for plant cell continuous culture |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105602847A (en) * | 2016-03-18 | 2016-05-25 | 南京晓庄学院 | Centrifugal settling-type bioreactor for continuous culture of plant cells |
| 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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2016
- 2016-03-18 CN CN201620209653.5U patent/CN205501326U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105602847A (en) * | 2016-03-18 | 2016-05-25 | 南京晓庄学院 | Centrifugal settling-type bioreactor for continuous culture of plant cells |
| 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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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160824 Termination date: 20200318 |
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| CF01 | Termination of patent right due to non-payment of annual fee |