CN100360657C - Horizontal drum type bioreactor - Google Patents
Horizontal drum type bioreactor Download PDFInfo
- Publication number
- CN100360657C CN100360657C CNB2005100432010A CN200510043201A CN100360657C CN 100360657 C CN100360657 C CN 100360657C CN B2005100432010 A CNB2005100432010 A CN B2005100432010A CN 200510043201 A CN200510043201 A CN 200510043201A CN 100360657 C CN100360657 C CN 100360657C
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- Prior art keywords
- urceolus
- cells
- inner core
- cylinder
- culture
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- 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.)
- Expired - Fee Related
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- 210000004027 cell Anatomy 0.000 claims abstract description 37
- 239000000463 material Substances 0.000 claims abstract description 12
- 238000004113 cell culture Methods 0.000 claims abstract description 10
- 241001465754 Metazoa Species 0.000 claims abstract description 7
- 230000001105 regulatory effect Effects 0.000 claims abstract description 7
- 241000700605 Viruses Species 0.000 claims abstract description 5
- 210000004748 cultured cell Anatomy 0.000 claims abstract description 4
- 238000000855 fermentation Methods 0.000 claims abstract description 4
- 230000004151 fermentation Effects 0.000 claims abstract description 4
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 4
- 235000015097 nutrients Nutrition 0.000 claims description 16
- 239000011229 interlayer Substances 0.000 claims description 4
- 230000000813 microbial effect Effects 0.000 claims description 2
- 210000002435 tendon Anatomy 0.000 claims description 2
- 244000005700 microbiome Species 0.000 abstract description 4
- 238000007789 sealing Methods 0.000 abstract description 4
- 239000000126 substance Substances 0.000 abstract description 3
- 238000004114 suspension culture Methods 0.000 abstract description 3
- 241000932156 Proteinus Species 0.000 abstract 1
- 230000000712 assembly Effects 0.000 abstract 1
- 238000000429 assembly Methods 0.000 abstract 1
- 230000002093 peripheral effect Effects 0.000 abstract 1
- 230000029219 regulation of pH Effects 0.000 abstract 1
- 239000013589 supplement Substances 0.000 abstract 1
- 239000000725 suspension Substances 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000010008 shearing Methods 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000005096 rolling process Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 230000008676 import Effects 0.000 description 4
- 238000013019 agitation Methods 0.000 description 3
- 229910001220 stainless steel Inorganic materials 0.000 description 3
- 239000010935 stainless steel Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241001062009 Indigofera Species 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 210000004102 animal cell Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000000254 damaging effect Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 238000010907 mechanical stirring Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000003466 welding Methods 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000005201 scrubbing Methods 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a bioreactor used for animal and plant cell culture or microorganism fermentation. The bioreactor is mainly composed of an inner rotatable cylinder of which the peripheral wall are provided with micropores, an outer cylinder with a constant temperature jacket, and relevant assemblies and attachments. The present invention adopts a horizontal double-layer rotatable cylinder type sealing structure, the inner rotatable cylinder is used as a culture tank for containing cells and microcarriers, and the outer fixed cylinder is used for providing a substance living environment for cells. The supplement of materials, the regulation of pH and the replacement of culture solutions are firstly carried out in the outer cylinder or a regulating tank, the culture solutions are filled in the inner cylinder and used by cells. The cultured cells can not pass through the micropores of the wall of the inner cylinder, and protein or viruses generated by the cells can pass through the micropores and enter the outer cylinder to be collected on line. The present invention is suitable for the batch type, semi-continuous or continuous microcarrier culture of attached cells as well as used for the suspension culture of microorganisms or suspension cells.
Description
Technical field
The present invention relates to a kind of bio-reactor that is used to carry out animal and plant cell cultures or microbial fermentation.
Background technology:
On engineering science, get used to the container that is used for microorganisms such as culturing bacterium, yeast is called " fermentor tank " (Fermentor), and the container of cultivating animal and plant cells is called " bio-reactor " (bioreactor).Because purpose of the present invention mainly is the high-density vitro culture that is used for zooblast, may be used for the cultivation of vegetable cell or bacterium simultaneously, so be referred to as " bio-reactor " in the presents.
Along with the continuous development of biotechnology industry, press for and solve the bio-reactor that is used for microorganism and the cultivation of animal and plant cell large scale.Once adopted the mechanical stir-reactor (also claiming retort) of chemical industry production unit improvement in the past.Because the shearing force of this class mechanical agitation type retort in mixing the material process be bigger, when being used for animal cell culture, because of zooblast do not have cell walls protection, can not tolerate reasons such as violent stirring and ventilation, fragile by cultured cells.Formed bubble also can produce damaging action to sensitive cells during the agitator high speed rotating.Be to reduce the shearing force of propeller-blade pair cell and promote the mass transfer ability of reactor, for traditional mechanical stirring mode of reactor with pass gas, heat transfer, mass transfer method and also carry out many improvement.Design: slurry formula, bar type, ship propulsion slurry, oblique blade or sail shape mechanical stirrer and double vibrations taper hole sieve plate stirred reactor, cage aeration-agitation reactor, double-deck cage aeration-agitation reactor etc.The reactor that wherein is primarily aimed at the animal cell culture invention has: rolling bottle reactor, fluidized-bed reactor, hollow fiber reactor, film reactor, air-lift unit formula reactor etc.
Because various type of cells survival condition, these equipment do not solve fundamentally still that the stirring shearing force that the animal and plant cell large scale cultivate to exist is big, material transmits inhomogeneous, device interior buildup and the problem at dead angle, and carry out problems such as continuous production that high-density, large scale culturing face and online collection product.
Summary of the invention:
Be used for the problem that stirring shearing force damaging cells, material that animal and plant cell high-density culture exists transmit inhomogeneous, device interior buildup and dead angle for overcoming existing equipment, and carry out the problems such as online collection product that large scale culturing faces.The invention provides that a kind of whipping performance uniqueness, mechanical stirring shearing force are little, material mass transfer process cell growth is disturbed less, can carry out highdensity substep or continuous flow cell is cultivated, and bio-reactor that can online collection product.The microcarrier that the present invention both can be used for suspension culture of cells, also can be used for attached cell is cultivated.
Technical scheme: adopt the double-deck drum-type sealed structure of horizontal rolling, the inner core of the rotation of can rolling is as the culture tank of holding cell and microcarrier, and the fixed urceolus provides necessary physical environment for cells survival.In the time of need adding material, regulate pH or change nutrient solution, can in urceolus or in regulating tank, carry out.There is network of micropores to separate between the inner/outer tube, can not be passed through micropore by cultured cells, and the protein of cells produce or virus can be by network of micropores by online results.
Technical characterstic:
1, this bio-reactor is owing to adopted double-layer horizontal rolling drum-type structure, when carrying out cell cultures with it, the microcarrier of viable cell or inoculating cell is to be grown in the inner core, and the substratum in the urceolus just provides the indirect environment-guarantee of growth metabolism for cell.Cell in the inner core carries out the exchange of nutrient and metabolite by the interior substratum of micropore on the inner tube wall (or screen cloth) and urceolus.When needs are added material, oxygen or are regulated potential of hydrogen in substratum, both can in urceolus, directly carry out, also nutrient solution can be drawn out to other regulating tank internal recycle from urceolus and carry out.Lowered the influence of operation factors to culture system.
When 2, this reactor was used for the microcarrier suspension culture of genetic engineering bacterium or zooblast, the macro-molecular protein that fermentation using bacteria produces or the virus of cell cultures can constantly be transferred in the nutrient solution of urceolus by the inner core micro-strainer.In not influencing tube, under the situation of bacterium or cell continued growth, can product be collected timely, realize semicontinuous batch of formula cultivation or cultured continuously by collecting the urceolus nutrient solution.
3, this reactor adopts horizontal drum type to stir the mass transfer mode, can eliminate buildup or dead angle, realizes the high efficient mixed of material and microcarrier.Also lowered simultaneously the injury effect of the shearing force pair cell of propeller agitator.
4, the gas of this reactor mix to be supplied with and directly not to be carried out in cell culture system, and this helps being avoided the injury effect of the surface tension pair cell of bubble.
5, cultivating bacterium or the cell that needs when finishing in the surge drum, can behind the emptying substratum, directly improve the inner core rotating speed, get rid of filter and reclaim cell.Can also utilize the filter effect of getting rid of of inner core, pair cell carries out the repetitive scrubbing processing in tube.
6, this reactor also have make simple, can sterilization system on line, carry out characteristics such as scale amplification easily.
Below in conjunction with accompanying drawing implementation step of the present invention is described further:
Description of drawings:
Accompanying drawing 1 is a structural representation of the present invention
Accompanying drawing 2 is inner tube structure figure of the present invention
Accompanying drawing 3 is outer tube structure figure of the present invention
Implementation step:
In accompanying drawing 1, this bio-reactor is mainly by rotatable inner core (1) with have the urceolus (2) of constant temperature jacket and antinose-dive leg (3) and annex (4) are formed.
In accompanying drawing 2 illustrated embodiments, inner core (1) is formed (accompanying drawing two) by inner core cylinder (1-1) and inner core bonnet (1-2), inner core protecgulum (1-3), inner core tripod assembly (1-4) riveted joint (or welding).Be covered with small hole on inner core cylinder (1-1) perisporium, the diameter 0.1-9000 micron in hole, the distance between the Kong Yukong is the 50-9000 micron.Inner core (1) main body is selected the punching press of 0.5-8 mm thick stainless steel plate for use, is twisted, welds, polishes and make, and it has the local location of micropore also can select 50-800 purpose sintered stainless steel screen making for use.
At the inwall of inner core (1) along axis to three to six convex tendons (1-1-1) that slight projection is arranged, be how much symmetries and split, the effect of rolling of culture when rotating in order to increase inner core.On the outer wall of inner core cylinder (1-1),, be how much symmetries and split along three to six put sinks (1-1-2) of axis to the perk of oriented inner core sense of rotation.During use, inner core slowly rotates, and when put sink leaves liquid level, can enter inner core by the micropore on the inner tube wall again with the part nutrient solution band in the urceolus from liquid level, to increase the fluid exchange of inside and outside tube.The wide ratio in the footpath of inner core is 0.1-20.The central part of inner core protecgulum have one suitably the circle of size connect the thing hole, be used in the inspection section situation or in tube, take a sample.
Inner core tripod assembly (1-4) by inner core tripod (1-4-1), main shaft (1-4-2), form.Inner core tripod assembly and inner core bonnet (1-2) riveted joint (or welding) are supported inner core (1) together.
In the embodiment shown in fig. 3, the urceolus circle (2-1) made by 1-10 mm thick stainless steel of urceolus (2), urceolus protecgulum (2-2), urceolus bonnet (2-3), urceolus circle front sealing flange (2-4), urceolus tripod assembly (2-5) etc. are formed (accompanying drawing three).The concentric chuck that urceolus circle (2-1) is made of jointly urceolus inner ring and urceolus outer ring.The inner ring and the outer ring of urceolus are welded on respectively on urceolus bonnet (2-3) and the urceolus front sealing flange (2-4) with concentric mode two ends, just constitute the urceolus main body.On the urceolus outer ring, be provided with interlayer recirculated water import and export.In this interlayer, feed flow media, can play the attemperation effect environment in the tube.Urceolus (2) is to be used for simultaneously main motor, well heater, temperature regulator, transmitter and the inside and outside exchange of substance passage of tube etc. being played a supportive role for reactor culturing room (inner core) provides isoperibol and holds nutrient solution.
In the centre of urceolus bonnet (2-3) circular hole is arranged, inner core main shaft (1-4-2) passes from the center hole of urceolus bonnet (2-3), and urceolus and inner core rotating shaft are linked up, and inner/outer tube is become one.The belt pulley or the gear that connect main motor are installed, by the main electric motor driving inner core rotation of adjustable speed on the main shaft.
In the position of urceolus circle near the urceolus protecgulum, all be provided with the passage (2-6) that several connect urceolus circle (2-1) up and down, be used to install sterilisation vapour import and export valve, material import and export valve, temperature control well heater, annexes such as outer cylinder sensor.
Urceolus protecgulum (2-2) is to engage with the anterior method indigo plant of urceolus circle (2-4) by method is blue, safety relief valve (2-2-1) is installed, withstand voltage glass observation window (2-2-2), thieff hatch (2-2-3), inner core sensor stand (2-2-4) on the urceolus protecgulum.After urceolus protecgulum (2-2) is by method indigo plant and urceolus sealing engagement, just can be used for cell cultures.If close relevant material import and export valve and thieff hatch, open internal heater or in reactor, feed water vapour, can carry out the sterilization system on line of reactor.
When using this bio-reactor to carry out cell cultures, the microcarrier that an amount of pre-treatment is good is placed in the inner core of reactor, all adds nutrient solution in inner/outer tube.The dosage of nutrient solution is with when inner core rotates, and the liquid in the inner core can not spill urceolus for well.Inner core slowly rotates during cultivation, and microcarrier or cell in the tube constantly stir with the rotation of inner core.Nutrient solution in the inner core is by the micropore on the inner tube wall, exchanges with nutrient solution in the urceolus.Because the aperture of micropore is little, cell or microcarrier can not enter urceolus by micropore.Guarantee the homo(io)thermism of reactor by the temperature of recirculated water in the external thermostatted adjusting urceolus interlayer.When the nutrient solution in the urceolus need replenish nutrient or regulate potential of hydrogen, nutrient solution is incorporated into from urceolus adjusts the regulating tank and add, recycle in the blowback reactor again by peristaltic pump then.Carry out Continuous Flow if desired and add the formula cultivation, initiate nutrient solution directly can be added inner core, the quantity discharged of control urceolus makes inflow, effusive liquid reach balance.Select the inner core of suitable big or small micropore, can carry out the microcarrier culturing cell, and successive is collected products such as monoclonal antibody or results virus from urceolus liquid at inner core.
Claims (2)
1, a kind of horizontal drum type bioreactor that is used to carry out animal and plant cell cultures or microbial fermentation, adopt double-deck drum-type sealed structure, it is characterized in that: this device is the rolled rotation inner core (1) that micropore is arranged by perisporium, fixed has interlayer urceolus (2) and antinose-dive leg (3) and annex (4) to form, the tube circle inwall of inner core (1) has convex tendon, outer wall has put sink, perisporium has the micropore of diameter 0.1-9000 micron, the protecgulum middle part has circle to connect the thing hole, inner core is as the culture tank of holding cell and microcarrier, urceolus provides the material living environment for cell, need add material, exchange back inner core again after in urceolus or regulating tank, carrying out earlier when regulating pH and changing nutrient solution, cultured cells can not be overflowed by micropore, and the protein of cells produce or virus can enter urceolus by online results by micropore.
2, according to the described bio-reactor of claim 1, it is characterized in that the protecgulum of reactor urceolus (2) is provided with observation window, thieff hatch, and the travel(l)ing rest that interior cylinder sensor is installed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100432010A CN100360657C (en) | 2005-03-30 | 2005-03-30 | Horizontal drum type bioreactor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100432010A CN100360657C (en) | 2005-03-30 | 2005-03-30 | Horizontal drum type bioreactor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1687386A CN1687386A (en) | 2005-10-26 |
| CN100360657C true CN100360657C (en) | 2008-01-09 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100432010A Expired - Fee Related CN100360657C (en) | 2005-03-30 | 2005-03-30 | Horizontal drum type bioreactor |
Country Status (1)
| Country | Link |
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| CN (1) | CN100360657C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805695B (en) * | 2010-01-29 | 2012-06-13 | 福尔生物制药股份有限公司 | Oblique rotational gravity mixed biological reaction equipment |
| CN101805694B (en) * | 2010-01-29 | 2012-07-04 | 福尔生物制药股份有限公司 | Gravity mixing bioreactor obliquely rotating along central shaft of culture vessel |
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101191154B (en) * | 2006-11-29 | 2010-05-12 | 中国科学院过程工程研究所 | Revolving drum type bioreactor used for microorganism lixiviation |
| CN106591102A (en) * | 2017-02-05 | 2017-04-26 | 西北农林科技大学 | Vertical rotary-drum-type biogas liquid in-situ quality-improving device and method |
| CN106701545A (en) * | 2017-02-11 | 2017-05-24 | 西北农林科技大学 | Horizontal rotary cylinder type biogas slurry in-situ quality-improving device and method |
| CN110157617B (en) * | 2019-04-29 | 2021-07-16 | 重庆大学附属肿瘤医院 | Cervical cancer cell three-dimensional culture device and its application |
| CN111530897B (en) * | 2020-05-25 | 2023-10-10 | 上海巷西环境科技有限公司 | A vacuum drum type food waste treatment device and its treatment technology |
| CN113699021A (en) * | 2021-07-30 | 2021-11-26 | 广西壮族自治区农业科学院 | Constant-temperature strain expanding culture separating tank |
-
2005
- 2005-03-30 CN CNB2005100432010A patent/CN100360657C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805695B (en) * | 2010-01-29 | 2012-06-13 | 福尔生物制药股份有限公司 | Oblique rotational gravity mixed biological reaction equipment |
| CN101805694B (en) * | 2010-01-29 | 2012-07-04 | 福尔生物制药股份有限公司 | Gravity mixing bioreactor obliquely rotating along central shaft of culture vessel |
| US11299700B1 (en) | 2021-02-19 | 2022-04-12 | Acequia Biotechnology, Llc | Bioreactor containers and methods of growing hairy roots using the same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1687386A (en) | 2005-10-26 |
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Application publication date: 20051026 Assignee: Jiangsu Yanshen Biology Technology Co., Ltd. Assignor: Han Yipeng Contract record no.: 2010320000288 Denomination of invention: Horizontal drum type bioreactor Granted publication date: 20080109 License type: Exclusive License Record date: 20100323 |
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Granted publication date: 20080109 Termination date: 20120330 |