CN104862265A - Plant cell entrapping device and entrapping method - Google Patents
Plant cell entrapping device and entrapping method Download PDFInfo
- Publication number
- CN104862265A CN104862265A CN201510225327.3A CN201510225327A CN104862265A CN 104862265 A CN104862265 A CN 104862265A CN 201510225327 A CN201510225327 A CN 201510225327A CN 104862265 A CN104862265 A CN 104862265A
- Authority
- CN
- China
- Prior art keywords
- filter membrane
- retort
- nutrient solution
- liquid pump
- plant cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
Discloses is a plant cell entrapping device. The plant cell entrapping device comprises a reaction tank, an upper filter membrane, a lower filter membrane, a double-way liquid pump and a waste liquid outlet. A plant cell entrapping method based on the plant cell entrapping device includes that a plant cell culture liquid is added into the reaction tank, the double-way liquid pump is started to alternately rotate, the culture liquid flows circularly alternately in different directions in the reaction tank, and efficient entrapping and separating of cells and the culture liquid can be achieved. The device is added to a plant cell culture reactor, then continuous perfusion culture can be performed, bottleneck problems in plant cell continuous culturing are solved by the simple device, and use value is great.
Description
Technical field
The present invention relates to biotechnology, particularly a kind of vegetable cell device for trapping and retain method.
Technical background
Utilize bio-reactor to carry out plant cell suspension culture and not only can produce many secondary metabolites with important value, and excessively excavating of wild resource can be alleviated.But the middle secondary metabolite that vegetable cell produces in culturing process accumulation in nutrient solution can suppress the further synthesis of product.Vegetable cell continuous perfusion culture constantly takes to outside reactor by material cell can be discharged into inside substratum, efficiently solves the problem of cell brown stain and product feedback inhibition.In vegetable cell continuous perfusion culture process, the separation efficiency of cell and nutrient solution is a bottle stem technology in this field.Conventional cell retention method has membrane filtration, centrifugal settling and gravity settling etc.Membrane filtration easily blocks, so be mainly used in zooblast without retaining, centrifugal settling can cause damage to the vegetable cell of sensitivity, and gravity settling efficiency is too low.
Summary of the invention
Object of the present invention, exactly in order to overcome above-mentioned prior art Problems existing, providing a kind of vegetable cell device for trapping and retaining method.
In order to realize object of the present invention, present invention employs following technical scheme: a kind of vegetable cell device for trapping, comprise retort, retort is provided with upper opening and lower openings, upper opening is provided with top filter membrane, lower openings is provided with bottom filter membrane, between upper opening and lower openings, is connected with a two-way liquid pump by pipeline, the pipeline between two-way liquid pump and upper opening is also connected with waste liquid outlet arm.
Described top filter membrane and bottom filter membrane are stainless steel metal film, and membrane pore size is 250-300 micron.
The outlet of described waste liquid outlet arm is higher than upper opening.
Vegetable cell based on above-mentioned vegetable cell device for trapping retains method and is, plant cell culturing liquid is filled in retort, open two-way liquid pump, nutrient solution is allowed to replace circulation with different directions in retort, when nutrient solution in retort in a clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of bottom, when nutrient solution in retort with counter clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of top, two-way liquid pump alternate rotation, guarantee to have a filter membrane at least not blocked, thus realize cell and be separated with efficiently retaining of nutrient solution.
The alternate rotation cycle of described two-way liquid pump is 2-30 minute.
After described two-way liquid pump is opened, the apparent velocity that nutrient solution replaces circulation in retort is 2-10ml/ second.
Device of the present invention is attached to after on culture plant cell reactor, can continuous perfusion culture be carried out, solve with a simple assembly bottleneck problem faced in vegetable cell cultured continuously, there is very large utility value.
Accompanying drawing explanation
Fig. 1 is the structural representation of vegetable cell device for trapping,
Fig. 2 is the apparatus structure schematic diagram of embodiment 1.
Specific implementation method
See Fig. 1, vegetable cell device for trapping of the present invention, comprise retort 5, retort 5 is provided with upper opening and lower openings, upper opening is provided with top filter membrane 1, lower openings is provided with bottom filter membrane 2, between upper opening and lower openings, is connected with a two-way liquid pump 3 by pipeline, the pipeline between two-way liquid pump and upper opening is also connected with waste liquid outlet arm 4.The outlet of this waste liquid outlet arm is higher than upper opening.
Top filter membrane in the present invention and bottom filter membrane are stainless steel metal film, and membrane pore size is 250-300 micron.
Vegetable cell based on above-mentioned vegetable cell device for trapping retains method and is, plant cell culturing liquid is filled in retort, open two-way liquid pump, nutrient solution is allowed to replace circulation with different directions in retort, when nutrient solution in retort in a clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of bottom, when nutrient solution in retort with counter clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of top, two-way liquid pump alternate rotation, guarantee to have a filter membrane at least not blocked, thus realize cell and be separated with efficiently retaining of nutrient solution.
Wherein the alternate rotation cycle of two-way liquid pump is 2-30 minute, and the apparent velocity that nutrient solution replaces circulation in retort is 2-10ml/ second.
Embodiment 1
Vegetable cell device for trapping of the present invention is attached in an inner ring gas lift-type retort, and continuous concern cultivation has been carried out to licorice cell.
Fig. 2 is the apparatus structure schematic diagram of embodiment 1.Wherein: 1-top filter membrane, 2-bottom filter membrane, the two-way liquid pump of 3-, 4-waste liquid outlet arm, 5-retort, 6-oxygen probe, 7-pH pops one's head in, 8-liquid meter, 9-gas blowout head, 10-on-line monitoring device, 11-computer, 12-thief hole, 13-air strainer, 14-gas pump, the unidirectional liquid pump of 15-.
Carried out cultured continuously by the present embodiment to cultivation licorice cell, initial inoculation density is 5g (dry weight)/L-1, and nutrient solution is the MS substratum adding NAA and IAA, and Ventilation Rate is 0.3vvm, and irrigation rate is 10%, and culture temperature is 25 DEG C.Result shows, cell retention efficiency reaches 100%, and after 27 days, cell maximum biomass reaches 17.5g L-1.
The present invention's simple device, achieves the high efficiency separation of vegetable cell and nutrient solution, can be extended to industrial scale applications.
Claims (6)
1. a vegetable cell device for trapping, it is characterized in that: comprise retort, retort is provided with upper opening and lower openings, upper opening is provided with top filter membrane, lower openings is provided with bottom filter membrane, between upper opening and lower openings, be connected with a two-way liquid pump by pipeline, the pipeline between two-way liquid pump and upper opening is also connected with waste liquid outlet arm.
2. vegetable cell device for trapping as claimed in claim 1, it is characterized in that: described top filter membrane and bottom filter membrane are stainless steel metal film, membrane pore size is 250-300 micron.
3. vegetable cell device for trapping as claimed in claim 1, is characterized in that: the outlet of described waste liquid outlet arm is higher than upper opening.
4. retain method based on the vegetable cell of vegetable cell device for trapping described in claim 1, it is characterized in that: plant cell culturing liquid is filled in retort, open two-way liquid pump, nutrient solution is allowed to replace circulation with different directions in retort, when nutrient solution in retort in a clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of bottom, when nutrient solution in retort with counter clockwise direction circulation time, blocking is washed in nutrient solution to the cell on the filter membrane of top, two-way liquid pump alternate rotation, guarantee to have a filter membrane at least not blocked, thus realize cell and be separated with efficiently retaining of nutrient solution.
5. vegetable cell retains method as claimed in claim 4, it is characterized in that: the alternate rotation cycle of described two-way liquid pump is 2-30 minute.
6. vegetable cell retains method as claimed in claim 4, it is characterized in that: after described two-way liquid pump is opened, and the apparent velocity that nutrient solution replaces circulation in retort is 2-10ml/ second.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510225327.3A CN104862265B (en) | 2015-05-04 | 2015-05-04 | Plant cell device for trapping and retention method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510225327.3A CN104862265B (en) | 2015-05-04 | 2015-05-04 | Plant cell device for trapping and retention method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104862265A true CN104862265A (en) | 2015-08-26 |
| CN104862265B CN104862265B (en) | 2018-02-02 |
Family
ID=53908446
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510225327.3A Expired - Fee Related CN104862265B (en) | 2015-05-04 | 2015-05-04 | Plant cell device for trapping and retention method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104862265B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755146A (en) * | 2015-11-19 | 2017-05-31 | 中国石油化工股份有限公司 | A kind of method and device of the production long-chain biatomic acid that continuously ferments |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0336966A1 (en) * | 1987-09-07 | 1989-10-18 | Hitachi, Ltd. | Cell culture method and apparatus |
| CN101182457A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Device for high-density production of organic acid by combination of fibrous bed bioreactor and membrane separation technology |
-
2015
- 2015-05-04 CN CN201510225327.3A patent/CN104862265B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0336966A1 (en) * | 1987-09-07 | 1989-10-18 | Hitachi, Ltd. | Cell culture method and apparatus |
| CN101182457A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Device for high-density production of organic acid by combination of fibrous bed bioreactor and membrane separation technology |
Non-Patent Citations (3)
| Title |
|---|
| FUREY J ET AL: "A low-shear filtration system that inhibits filter-membrane fouling", 《GENETIC ENGINEERING NEWS》 * |
| KELLY W ET AL: "Understanding and modeling alternating tangential flow filtration for perfusion cell culture", 《BIOTECHNOL PROG》 * |
| SPECTRUM LABORATORIES,INC: "Conversion of bioreactors to continuous perfusion using hollow fiber cell separators", 《WWW.SPECTRUMLABS.COM/LIT/CPAG.PDF》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755146A (en) * | 2015-11-19 | 2017-05-31 | 中国石油化工股份有限公司 | A kind of method and device of the production long-chain biatomic acid that continuously ferments |
| CN106755146B (en) * | 2015-11-19 | 2020-04-10 | 中国石油化工股份有限公司 | Method and device for producing long-chain dicarboxylic acid by continuous fermentation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104862265B (en) | 2018-02-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN204727884U (en) | A kind of high-density culture is coupled the film bioreactor of pre-microalgae | |
| Zhuang et al. | A novel suspended-solid phase photobioreactor to improve biomass production and separation of microalgae | |
| Singh et al. | Harvesting of microalgal biomass | |
| Arcuri et al. | Ethanol production by immobilized cells of Zymomonas mobilis | |
| CN105861370A (en) | High-density culture and pre-harvesting method of microalgae | |
| CN102703535A (en) | New technology for producing acrylamide by using ceramic membrane bioreactor | |
| CN107376436A (en) | A kind of phytoplankton subsiding separator | |
| CN101130751B (en) | Continuous bottling and culturing device of plant cell stirring type bioreactor | |
| CN101693878B (en) | Microalgae concentrating and collecting method and device thereof | |
| Mehrotra et al. | Bioreactor technology for hairy roots cultivation | |
| CN102337200A (en) | High-efficiency microcarrier cell-culture medium separating device | |
| CN105602847A (en) | Centrifugal settling-type bioreactor for continuous culture of plant cells | |
| CN204325379U (en) | A kind of microcarrier and cell culture apparatus | |
| CN104862265A (en) | Plant cell entrapping device and entrapping method | |
| KR101663036B1 (en) | Divice for microalgae concentrating and hydrogen generating | |
| CN110220746A (en) | A kind of field planktonic organism quantitative collection bag and application method | |
| CN203960205U (en) | Cell settlement groove | |
| CN205501326U (en) | Centrifugal sedimentation formula bioreactor for plant cell continuous culture | |
| CN208485864U (en) | A kind of recirculation reactor collecting microdisk electrode and harvesting | |
| CN206666422U (en) | A kind of device of embrane method extraction humic acid | |
| CN103305418A (en) | Stem cell isolated culturing device | |
| CN107473494A (en) | The device and method of ammonia nitrogen in a kind of removal waste water | |
| GANI et al. | Cultivation system and harvesting techniques in Microalgae Biomass production | |
| CN107473383B (en) | Device and method for treating ammonia nitrogen wastewater | |
| CN203947105U (en) | A kind of cell dissociation equipment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180202 Termination date: 20200504 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |