CN108949558A - The method of roller hollow fiber reactor and Guava Leaf high cell densities culture for plant cell High Density Cultivation - Google Patents
The method of roller hollow fiber reactor and Guava Leaf high cell densities culture for plant cell High Density Cultivation Download PDFInfo
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- CN108949558A CN108949558A CN201810802922.2A CN201810802922A CN108949558A CN 108949558 A CN108949558 A CN 108949558A CN 201810802922 A CN201810802922 A CN 201810802922A CN 108949558 A CN108949558 A CN 108949558A
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- 238000001816 cooling Methods 0.000 claims description 3
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- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 claims description 3
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- 238000010008 shearing Methods 0.000 abstract description 13
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- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
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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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/10—Hollow fibers or tubes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/04—Plant cells or tissues
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Abstract
The invention discloses a kind of roller hollow fiber reactors for plant cell High Density Cultivation, including roller, hollow fiber conduit, culture solution circulation pipe and driving device for driving roller rotation, the roller is not connected to the external world, the hollow fiber conduit is fixedly arranged in the roller, the quantity of the hollow fiber conduit is at least one, the culture solution circulation pipe is connected with the lumen of hollow fiber conduit, and the culture solution circulation pipe is equipped with culture solution circulating pump.The present invention correspondingly provides a kind of method using above-mentioned roller hollow fiber reactor culture Guava Leaf cell.Apparatus of the present invention good mixing property, shearing force when cultivating cell is low and energy saving.In addition, apparatus of the present invention are flowed since roller rotates in band mobile shell fluid, its dissolved oxygen is agglomerated and improved to the tight agglomeration that can substantially reduce cell, improves the mass transfer permeability of film, is conducive to cell growth metabolism, is conducive to concentration cultivation.
Description
Technical field
The invention belongs to technical field of cell culture more particularly to a kind of reactors and Guava Leaf for cell culture
The method of cell culture.
Background technique
Guava plant is a kind of plant with good medicinal and nutritional values.But be mass produced with it valuable
When product, then exist crop cycle is long, take up a large area, recovery process long flow path, it is at high cost the disadvantages of.Guava plant cell
Large-scale culture, be realize guava valuable constituent efficiently, low cost, the effective way that utilizes of scale.
Culture plant cell has the characteristics that the period is long, cell anti-shear ability is weak, easy to reunite.Plant cell scale evaluation
Purpose be that production obtains valuable natural products, the i.e. metabolin of cell growth.The design of culture plant cell reactor and
Its process should consider to be conducive to cell growth, consider the accumulation and separation that are conducive to product again, also it be required to have
The characteristics such as suitable oxygen transmitting, good mobility, lower shearing force, low energy consumption.Currently, for the anti-of culture plant cell
Answering device mainly has stirring-type, non-stirred formula, plant cell immobilization reactor, membrane reactor etc..
Mechanical agitation type bioreactor has biggish opereating specification, mixability height, wide adaptability, is being mass produced
In be widely applied.But the shearing force generated in agitator tank is big, is easy to damage cell, directly affects growth and the generation of cell
It thanks, especially generating on secondary species influences greatly.So the plant cell sensitive to shearing force, traditional mechanical agitator tank is not
It is too applicable.For this purpose, improving to the whipped form of agitator tank, blade wheel structure and type, air sparger etc., make every effort to reduce producing
Raw shearing force, while meeting oxygen supply and mixed requirement, such as it can be adapted for shearing force using with the band-like impeller of double helix
Sensitive cell culture；Three-bladed propeller reactor (MRP) using micropore metal silk screen as air sparger can provide smaller
Shearing force and good oxygen supply and admixture, be better than six flat-blade turbine paddle reactors.
Relative to traditional stirring reactor, shearing force caused by non-stirred formula reactor is smaller, and structure is simple, compares
It is suitble to culture plant cell, main Types have bubbling style reactor, airlift reactor etc..Bubbling style reactor is generally preferred over
Mechanical stir-reactor, but since utilization rate of the bubbling style reactor to oxygen is lower, if with larger ventilatory capacity, larger bubble
Or will lead to higher shear power compared with high gas velocity, it is unfavorable for culture plant cell.Airlift reactor is widely used in plant cell
The research and production of culture are generally used for various plants cell suspension cultures or immobilized growing cells, but its operating flexibility is small,
Low gas velocity or when phase cell density is higher after incubation, mixed effect is poor, it is desirable that ratio of height to diameter H/D is big.If improving ventilatory capacity
It improves mixed effect, and a large amount of foams can be generated, be also easy to drive away the carbon dioxide etc. in culture solution, to plant cell growth
There is inhibition, in addition, it is also unfavorable to plant cell synthesis secondary metabolite to improve the excessively high dissolved oxygen of ventilatory capacity bring.
According to the characteristic of plant cell, it is thin that the novel reactor that many is different from traditional microbiological reactor is used for plant
Born of the same parents' researchs and produces, such as various immobilization plant cell reactors and membrane reactor.Surface of Plant callus cell Immobilized culture system
System avoids fluid flow forces or shearing force problem in conventional stirred-tank suspension culture, and the spy for promoting plant cell to agglomerate
Property, so that secondary metabolite is synthesized and is accumulated and increase, and systematic cultivation base exchange is simple, secondary species extracts easy.It plants
The cells are fixed on film for object cell membrane reactor (i.e. " quiet " type hollow fiber membrane reactor), culture medium loop under film
It circulates (i.e. the tube side of hollow fiber film tube), nutrition is expanded through membrane diffusion to cellular layer, secondary metabolites secretion through film
It is dissipated to culture medium.But immobilization plant cell reactor and membrane reactor be there are mass transfer velocity is slow and the unbalanced problem of mass transfer, when
When anchorage-dependent cells culture density constantly increases, mixing situation is deteriorated, and it is excessive that there are cells on film surface part or wall
A large amount of frothings are agglomerated and generated, causes the supply of cell condensation environmental interior nutriment and oxygen insufficient, leads to living cells
Amount and metabolite are reduced, and incubation is forced to stop when serious.
Therefore, research and develop a kind of membrane mass transfer permeability it is good, conducive to cell growth metabolism, to prevent cell transition from reuniting solidifying
Poly- novel cell cultivation reactor has a vast market foreground.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the shortcomings of to mention in background above technology and defect, to existing plant
The membrane reactor and its technique of object cell culture carry out synthesis improvement, provide that a kind of shearing is small, and low energy consumption, membrane mass transfer permeability
Well, conducive to cell growth metabolism, the roller hollow fiber reactor for preventing cell transition reunion from agglomerating, and a kind of kind is accordingly provided
The method of pomegranate leaf high cell densities culture.In order to solve the above technical problems, technical solution proposed by the present invention are as follows:
A kind of roller hollow fiber reactor for plant cell High Density Cultivation, including roller, hollow fiber conduit, training
Nutrient solution circulation pipe and driving device for driving roller rotation, the roller are not connected to extraneous, the hollow fibre
Dimension pipe is fixedly arranged in the roller, and the quantity of the hollow fiber conduit is at least one, the culture solution circulation pipe and hollow fibre
The lumen of dimension pipe is connected, and the culture solution circulation pipe is equipped with culture solution circulating pump.
In the above-mentioned roller hollow fiber reactor for plant cell High Density Cultivation, hollow fiber conduit be by poly- maple or
The semi-permeable membrane of the polymer composition of propylene, hydrone, nutriment, cell metabolite and gas can penetrate, oxygenous and battalion
The culture solution for supporting substance is flowed through from the lumen of hollow fiber conduit, and cell is grown in doughnut tube outer surface, and cell grows institute
Water, nutriment for needing etc. reach at cell since concentration difference can pass through hollow fiber conduit, the metabolite that cell generates
Permeable hollow fiber conduit arrival doughnut tube cavity is cultured liquid band and walks, and the nutrition between cell and culture solution exchanges more
Sufficiently efficiently, it is more conducive to the growth and breeding of cell.
Conventional " quiet " type hollow fiber reactor has some inherent defects: the phase is since cell density sharply increases after incubation
Greatly, it will lead to viscosity increase, mixed performance becomes poor, aggregation cohesion, so that dissolved oxygen is deteriorated in aggregate, because inside lacks
Oxygen and cause cell death.Roller hollow fiber reactor in the present invention is thin since the low sheraing rotation of roller can substantially reduce
The cohesion that closes the ranks of born of the same parents, can improve the mass transfer permeability of film, to be conducive to concentration cultivation.
In above-mentioned roller hollow fiber reactor, it is preferred that the roller hollow fiber reactor further includes that cell liquid follows
Endless tube, the cell liquid circulation pipe are connected with the shell chamber that hollow fiber conduit and inner wall of rotary drum are formed, the cell liquid circulation pipe
It is equipped with cell liquid circulating pump.Only routine " quiet " type hollow fiber reactor of the culture solution flowing in hollow fiber film tube,
Late stage of culture is brought a series of the problem of referring in backgrounds since cell density is excessive, and roller rotation can be very effective
These disadvantages of conventional " quiet " type hollow fiber reactor are solved, meanwhile, cell liquid circulation pipe is set, not only realizes cell liquid
It interior flowing under rollers roll and can be flowed with outer circulation, the tight agglomeration of cell is solidifying when can substantially reduce high density
It is poly-, and improve its dissolved oxygen and CO2, it is more conducive to cell growth metabolism, is conducive to concentration cultivation.In addition, setting is thin
Cytosol circulation pipe can also facilitate separates cell out of roller, achievees the effect that separate in culture, to realize continuously or semi-continuously
Operation.
In above-mentioned roller hollow fiber reactor, it is preferred that revolved equipped with roller in the centre of the both ends of the surface of the roller
Turn connecting shaft, the roller rotation connecting shaft by connector sealing, rotatable be sheathed on a fixed hollow sleeve
Guan Shang is equipped with the first connecting tube being connected with lumen and the second connecting tube being connected with shell chamber, institute in the hollow bushing
It states the first connecting tube and the second connecting tube is connected with culture solution circulation pipe and cell liquid circulation pipe respectively.The roller at roller both ends
When rotating connecting bushing on hollow bushing, roller rotates the connector between connecting shaft and hollow bushing and uses rotating machinery
End face seal, when roller is rotated around hollow bushing, extraneous gas containing bacterium also cannot be introduced into roller.
In above-mentioned roller hollow fiber reactor, it is preferred that the quantity of the hollow fiber conduit is more, the roller two
The first connecting tube in the hollow bushing at end protrudes into the roller, and the end for protruding into the first connecting tube of the roller is equal
Equipped with fixed disc, the more hollow fiber conduits are fixedly arranged in the roller by two fixed discs.In practical operation, if
Hollow fiber conduit is rotated together with roller, can be brought biggish shearing force (being equivalent to general blender stirring), and hollow fibre
(tube side) is rotated together with (shell chamber) in roller in dimension pipe, then two different fluids of tube side and shell chamber continuously cannot respectively enter and leave
The equipment, is unable to continuous production, can only Batch Process.Using said structure design of the invention, fixed disc will pass through first
Connecting tube is fixed in roller, and hollow fiber conduit will not be rotated synchronously with roller, and whole device only has cell culture fluid with rolling
Cylinder is rotated and is flowed, and the hollow fiber conduit of drums inside is always fixation, can both substantially reduce shearing force in this way to thin
The influence of born of the same parents can prevent the immobilized cell later period tightly packed and bring a series of problems again.
In above-mentioned roller hollow fiber reactor, it is preferred that be installed with the lamp for controlling light application time in the roller
Pipe, the fluorescent tube is also fixedly arranged between two fixed discs, and it is parallel with hollow fiber conduit and be in etc. to control the fluorescent tube
Side triangle Heterogeneous Permutation, the equilateral triangle Heterogeneous Permutation refer to that hollow fiber conduit is located at three vertex of equilateral triangle
On, fluorescent tube is located at the center of equilateral triangle.Be arranged fluorescent tube can conveniently regulating and controlling illumination and its time, improve plant cell growth generation
The validity that light source is obtained when thanking is conducive to plant cell to meet the needs of cell is to different illumination intensity and light application time
Growth.Illumination can be utilized using equilateral triangle Heterogeneous Permutation to greatest extent, and illumination patterns are more evenly, more rationally.
In above-mentioned roller hollow fiber reactor, it is preferred that the driving device include set on cylinder surface transfer roller with
And the transfer roller driving mechanism for driving the transfer roller rotation.
In above-mentioned roller hollow fiber reactor, it is preferred that the cell liquid circulation pipe is equipped with cell separation apparatus；Institute
It states culture solution circulation pipe and is equipped with the gas-liquid exchange device adjusted with carbon dioxide for realizing oxygen in culture solution, for removing
(the thin to plant of inhibition metabolite can be substantially reduced to the harmful substance that cell metabolism generates in culture solution by removing harmful substance
The feedback inhibition of intracellular growth) adsorption separation device and for adjusting culture solution constituent adjustment and temperature temperature control charging
Device；One is additionally provided on the culture solution circulation pipe, and for controlling, the culture solution does not enter or part enters the adsorbing separation
The culture solution branch of device, the entrance of the culture solution branch and the entrance of the adsorption separation device pass through a triple valve and institute
State the connection of culture solution circulation pipe, exit and the culture solution of the outlet of the culture solution branch in the adsorption separation device
Circulation pipe connection.Various works in culture solution can be achieved in the setting of gas-liquid exchange device, adsorption separation device and temperature control feeding device
The control of skill parameter (such as temperature, time, oxygen concentration, nutrient concentrations), to meet the needs of cell growth metabolism.Setting
Culture solution branch optionally enters culture solution or partially enters adsorption separation device, needs to remove the inhibition in culture solution
Property metabolite when, triple valve close culture solution branch, so that culture solution is entered adsorption separation device, conversely, triple valve close inhale
The entrance of attached separator returns to culture solution in culture solution circulation pipe along culture solution branch.
In above-mentioned roller hollow fiber reactor, it is preferred that the adsorption separation device is adsorption column, in the adsorption column
It is installed with absorbent particles, the absorbent particles are active carbon, aluminum oxide, surface through hydroxyl, carboxyl or amido modified
One or more of silica.
As a general technical idea, the present invention also provides a kind of sides that Guava Leaf high cell densities are continuously cultivated
Method, this method utilize above-mentioned roller hollow fiber reactor culture Guava Leaf cell, comprising the following steps:
(1) sterilization treatment and cooling are carried out to the roller hollow fiber reactor and culture solution；
(2) by the culture solution after sterilizing be added to it is sterilized after roller hollow fiber reactor in, and be inoculated with guava
Leaf cell opens circulating pump, opens driving device and drive to the doughnut tube outer surface of the roller hollow fiber reactor
Roller rotation, (liquid feeding and inoculation pass through the sealing cover mouth on roller stack shell)；
(3) highdensity Guava Leaf cell can be obtained in control condition of culture culture.
In the method for above-mentioned Guava Leaf high cell densities culture, it is preferred that the condition of culture control are as follows: cultivation temperature
It is 15-40 DEG C, intensity of illumination 2000-3000Lx, light application time 6-14h/d, drum rotation speed 5-50rpm, recyclegas stream
Fast (mixed gas of the carbon dioxide and oxygen used in gas-liquid exchange device) is 200-1000mL/min.
By utilization roller hollow fiber reactor culture Guava Leaf cell, it can be achieved that the big rule of Guava Leaf cell
Mould, High Density Cultivation, be realize in Guava Leaf valuable constituent efficiently, low cost, the effective way that utilizes of scale.
Compared with the prior art, the advantages of the present invention are as follows:
1, apparatus of the present invention good mixing property, shearing force when cultivating cell is low and energy saving, anti-with traditional air blow and agitation
Device is answered to compare, 2/3rds or more energy conservations, shear action can reduce 95%, have to change the rheological behavior of cell culture fluid and fit
The characteristics of Ying Xingqiang.
2, apparatus of the present invention are flowed since roller rotates in band mobile shell fluid, and the tight agglomeration that can substantially reduce cell is solidifying
Gather and improve its dissolved oxygen, improve the mass transfer permeability of film, is conducive to cell growth metabolism, is conducive to concentration cultivation.
3, culture solution is flowed through from the inner cavity of hollow fiber conduit in apparatus of the present invention, and cell is located at the appearance of hollow fiber conduit
Face can increase the contact area of cell and culture solution, and more sufficiently efficiently, the growth for being more conducive to cell is numerous for nutrition exchange
It grows.
4, Guava Leaf cell culture processes of the invention are simple, efficient, are applicable to extensive continuous, industrial metaplasia
It produces.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is the present invention
Some embodiments for those of ordinary skill in the art without creative efforts, can also basis
These attached drawings obtain other attached drawings.
Fig. 1 is the structural schematic diagram in embodiment for the roller hollow fiber reactor of plant cell High Density Cultivation.
Fig. 2 is the Section View in the face A-A in Fig. 1.
Marginal data:
1, roller；101, shell chamber；2, hollow fiber conduit；201, lumen；3, culture solution circulation pipe；31, gas-liquid exchange device；
32, adsorption separation device；33, temperature control feeding device；34, culture solution branch；4, driving device；41, transfer roller；42, transfer roller drives
Mechanism；5, culture solution circulating pump；6, cell liquid circulation pipe；61, cell separation apparatus；7, cell liquid circulating pump；8, roller rotates
Connecting shaft；9, connector；10, hollow bushing；11, the first connecting tube；12, the second connecting tube；13, fixed disc；14, fluorescent tube；
15, tube power line；16, sealing cover mouth.
Specific embodiment
To facilitate the understanding of the present invention, the present invention is made below in conjunction with Figure of description and preferred embodiment more complete
Face meticulously describes, but the protection scope of the present invention is not limited to the following specific embodiments.
Unless otherwise defined, all technical terms used hereinafter and the normally understood meaning of those skilled in the art
It is identical.Technical term used herein is intended merely to the purpose of description specific embodiment, is not intended to the limitation present invention
Protection scope.
Unless otherwise specified, various raw material, reagent, the instrument and equipment etc. used in the present invention can pass through city
Field is commercially available or can be prepared by existing method.
Embodiment:
As shown in Figure 1, the roller hollow fiber reactor for plant cell High Density Cultivation of the present embodiment, including rolling
Cylinder 1, hollow fiber conduit 2, culture solution circulation pipe 3 and the driving device 4 for driving roller 1 to rotate, roller 1 do not connect with the external world
Logical, hollow fiber conduit 2 is fixedly arranged in roller 1, and the quantity of hollow fiber conduit 2 is at least one, culture solution circulation pipe 3 and hollow fibre
The lumen 201 of dimension pipe 2 is connected, and culture solution circulation pipe 3 is equipped with culture solution circulating pump 5.
In the present embodiment, roller hollow fiber reactor further includes cell liquid circulation pipe 6, cell liquid circulation pipe 6 with it is hollow
Fibre pipe 2 is connected with the shell chamber 101 that 1 inner wall of roller is formed, and cell liquid circulation pipe 6 is equipped with cell liquid circulating pump 7.
In the present embodiment, the centre of the both ends of the surface of roller 1 is equipped with roller and rotates connecting shaft 8, and roller rotates connecting shaft 8
By a connector 9 sealing (passing through rotating machinery end face seal), rotatable it is sheathed on a fixed hollow bushing 10
On, the first connecting tube 11 being connected with lumen 201 and the second connecting tube being connected with shell chamber 101 are equipped in hollow bushing 10
12, the first connecting tube 11 and the second connecting tube 12 are connected with culture solution circulation pipe 3 and cell liquid circulation pipe 6 respectively.
In the present embodiment, the quantity of hollow fiber conduit 2 is more, the first connection in the hollow bushing 10 at 1 both ends of roller
Pipe 11 protrudes into roller 1, and the end for protruding into the first connecting tube 11 of roller 1 is equipped with fixed disc 13, more hollow fibres
Dimension pipe 2 is fixedly arranged in roller 1 by two fixed discs 13.
In the present embodiment, (the tube power line 15 of fluorescent tube 14 of fluorescent tube 14 for controlling light application time is installed in roller 1
By hollow bushing 10 introduce roller 1 inside), fluorescent tube 14 is also fixedly arranged between two fixed discs 13, and control fluorescent tube 14 with
Hollow fiber conduit 2 is parallel and Heterogeneous Permutation in equilateral triangle, equilateral triangle Heterogeneous Permutation refer to that hollow fiber conduit 2 is located at
On three vertex of side triangle, fluorescent tube 14 is located at the center of equilateral triangle.With three hollow fiber conduits 2 and a fluorescent tube 14
For, layout drawing is as shown in Figure 2.
In the present embodiment, driving device 4 includes set on the transfer roller 41 on 1 surface of roller and for driving transfer roller 41 to rotate
Transfer roller driving mechanism 42.Driving device 4 in the present embodiment be not limited to above-mentioned restriction transfer roller 41 and transfer roller driving mechanism 42, one
As common driving method, as long as the driving effect being able to satisfy in the present embodiment.
In the present embodiment, cell liquid circulation pipe 6 is equipped with cell separation apparatus 61；Culture solution circulation pipe 3 is equipped with and is used for
Realize oxygen and the gas-liquid exchange device 31 of carbon dioxide adjusting in culture solution, for removing cell metabolism generation in culture solution
The adsorption separation device 32 of harmful substance and temperature control feeding device 33 for adjusting culture solution constituent adjustment and temperature；Culture
It is additionally provided with one on liquid circulation pipe 3 and does not enter or for controlling culture solution partially into the culture solution branch of adsorption separation device 32
34, the entrance of culture solution branch 34 is connect by a triple valve with culture solution circulation pipe 3 with the entrance of adsorption separation device 32, is trained
The outlet of nutrient solution branch 34 is connected in the exit of adsorption separation device 32 with culture solution circulation pipe 3.
In the present embodiment, adsorption separation device 32 is adsorption column, is installed with absorbent particles, absorbent particles in adsorption column
It is active carbon, aluminum oxide, surface through one or more of hydroxyl, carboxyl or amido modified silica.
The device of the present embodiment have the advantage that the device of 1, the present embodiment when cultivating cell good mixing property, cut
Shear force is low and energy saving, and compared with traditional air blow and agitation reactor, 2/3rds or more energy conservations, shear action can reduce 95%,
Have the characteristics that change the rheological behavior of cell culture fluid adaptable.2, the device of the present embodiment is due to the rotation band of roller 1
Its dissolved oxygen is agglomerated and improved to flowing, the tight agglomeration that can substantially reduce cell in 101 fluid of mobile shell, and the mass transfer for improving film is penetrating
Property, it is conducive to cell growth metabolism, is conducive to concentration cultivation.3, in the device of the present embodiment culture solution from hollow fiber conduit
It is flowed through in 2 lumen 201, cell is located at the outer surface of hollow fiber conduit 2, can increase the contact area of cell and culture solution, seeks
Feeding exchange more sufficiently efficiently, is more conducive to the growth and breeding of cell.
The present embodiment also provides a kind of method of Guava Leaf high cell densities culture, and this method utilizes above-mentioned for plant
The roller hollow fiber reactor culture Guava Leaf cell of high cell densities culture, comprising the following steps:
(1) 125 DEG C of steam are accessed, steam sterilizing 15min is carried out to the roller hollow fiber reactor of 5L, then by 4L plant
The pH value of the general MS culture medium of cell is adjusted to 5.5-6.5, is loaded into autoclave sterilization pot and sterilizes, sterilising conditions are
0.1MPa, 121.5 DEG C, 15-30min take out cooling after sterilizing is good；
(2) the sealing cover mouth 16 that the culture medium after sterilizing directly passes through roller 1 is added, and be inoculated with by pectinase treatment-
For the Guava Leaf cell that Centrifugation-Filtration obtains to the outer wall of hollow fiber conduit 2, inoculum concentration is 1 × 105Cells/mL is opened
Each circulating pump and driving device 4 drive roller 1 to rotate；
(3) control condition of culture is as follows: cultivation temperature is 28 ± 1 DEG C, and intensity of illumination is 2000-3000Lx (above range
), light application time 12h/d, the revolving speed 5-50rpm (above range) of roller 1, recyclegas flow velocity are 600mL/
Min, gap to culture solution extra-nutrition substance and remove the inhibitory substance in culture solution for a period of time, and height can be obtained in culture
The Guava Leaf cell of density (culture midway may cross cell liquid circulation pipe 6 and collect highdensity Guava Leaf cell).
In the method for the Guava Leaf cell culture of the present embodiment, one subculture of supplement was observed and adjusted every 4 days, is mended
It fills the nutriments such as hormone, microelement and removes inhibitory substance (culture solution passed through an adsorption separation device every 4 days),
After cultivating 3 weeks, Guava Leaf cell density is 2.3 × 108cells/mL.One subculture of supplement was observed and adjusted every 1 day,
The nutriments such as hormone supplemented, microelement simultaneously remove inhibitory substance (culture solution every other day pass through adsorbing separation dress
Set), after culture 3 weeks, Guava Leaf cell density is 7.8 × 108cells/mL.It was observed every 1 day and adjusts the primary training of supplement
Base is supported, the nutriments such as hormone supplemented, microelement but is not removed inhibitory substance (culture solution is without adsorption separation device),
After cultivating 3 weeks, Guava Leaf cell density is 3.4 × 108cells/mL。
Claims (10)
1. a kind of roller hollow fiber reactor for plant cell High Density Cultivation, which is characterized in that including roller (1),
Hollow fiber conduit (2), culture solution circulation pipe (3) and the driving device (4) for driving the roller (1) to rotate, the rolling
Cylinder (1) is not connected to the external world, and the hollow fiber conduit (2) is fixedly arranged in the roller (1), the number of the hollow fiber conduit (2)
Amount is at least one, and the culture solution circulation pipe (3) is connected with the lumen (201) of hollow fiber conduit (2), and the culture solution follows
Endless tube (3) is equipped with culture solution circulating pump (5).
2. the roller hollow fiber reactor according to claim 1 for plant cell High Density Cultivation, feature exist
In the roller hollow fiber reactor further includes cell liquid circulation pipe (6), the cell liquid circulation pipe (6) and doughnut
Pipe (2) is connected with the shell chamber (101) that roller (1) inner wall is formed, and the cell liquid circulation pipe (6) is equipped with cell liquid circulating pump
(7)。
3. the roller hollow fiber reactor according to claim 2 for plant cell High Density Cultivation, feature exist
In the centre of the both ends of the surface of the roller (1) is equipped with roller rotation connecting shaft (8), and roller rotation connecting shaft (8) is logical
Cross a connector (9) sealing, it is rotatable be sheathed on a fixed hollow bushing (10), in the hollow bushing (10)
It is described equipped with the first connecting tube (11) being connected with lumen (201) and the second connecting tube (12) being connected with shell chamber (101)
First connecting tube (11) and the second connecting tube (12) are connected with culture solution circulation pipe (3) and cell liquid circulation pipe (6) respectively.
4. the roller hollow fiber reactor according to claim 3 for plant cell High Density Cultivation, feature exist
In the quantity of the hollow fiber conduit (2) is more, the first connecting tube in the hollow bushing (10) at roller (1) both ends
(11) it protrudes into the roller (1), and the end for protruding into the first connecting tube (11) of the roller (1) is equipped with fixed disc
(13), the more hollow fiber conduits (2) are fixedly arranged in the roller (1) by two fixed discs (13).
5. the roller hollow fiber reactor according to claim 4 for plant cell High Density Cultivation, feature exist
In the interior fluorescent tube (14) being installed with for controlling light application time of the roller (1), the fluorescent tube (14) is also fixedly arranged on described in two
Between fixed disc (13), and the fluorescent tube (14) Heterogeneous Permutation parallel and in equilateral triangle with hollow fiber conduit (2) is controlled,
The equilateral triangle Heterogeneous Permutation refers to that hollow fiber conduit (2) is located on three vertex of equilateral triangle, fluorescent tube (14) position
In the center of equilateral triangle.
6. the roller doughnut according to any one of claims 1-5 for plant cell High Density Cultivation reacts
Device, which is characterized in that the driving device (4) includes set on the transfer roller (41) on roller (1) surface and for driving described turn
The transfer roller driving mechanism (42) of roller (41) rotation.
7. being reacted according to any one of claim 2-5 for the roller doughnut of plant cell High Density Cultivation
Device, which is characterized in that the cell liquid circulation pipe (6) is equipped with cell separation apparatus (61)；On the culture solution circulation pipe (3)
Equipped with the gas-liquid exchange device (31) adjusted for realizing oxygen in culture solution and carbon dioxide, for removing cell in culture solution
The adsorption separation device (32) and the temperature control for adjusting culture solution constituent adjustment and temperature for being metabolized the harmful substance generated add
Expect device (33)；One is additionally provided on the culture solution circulation pipe (3), and for controlling, the culture solution does not enter or part enters institute
State the culture solution branch (34) of adsorption separation device (32), the entrance and the adsorption separation device of the culture solution branch (34)
(32) entrance is connect by a triple valve with the culture solution circulation pipe (3), and the outlet of the culture solution branch (34) is in institute
The exit for stating adsorption separation device (32) is connected to the culture solution circulation pipe (3).
8. the roller hollow fiber reactor according to claim 7 for plant cell High Density Cultivation, feature exist
In the adsorption separation device (32) is adsorption column, is installed with absorbent particles in the adsorption column, the absorbent particles are
Active carbon, aluminum oxide, surface are through one or more of hydroxyl, carboxyl or amido modified silica.
9. a kind of method of Guava Leaf high cell densities culture, which is characterized in that using described in any one of claim 1-8
The roller hollow fiber reactor culture Guava Leaf cell for plant cell High Density Cultivation, comprising the following steps:
(1) sterilization treatment and cooling are carried out to the roller hollow fiber reactor and culture solution；
(2) by the culture solution after sterilizing be added to it is sterilized after roller hollow fiber reactor in, and it is thin to be inoculated with Guava Leaf
Born of the same parents open circulating pump, open driving device (4) band to hollow fiber conduit (2) outer surface of the roller hollow fiber reactor
Dynamic roller rotation；
(3) condition of culture culture is controlled, highdensity Guava Leaf cell can be obtained.
10. the method for high density Guava Leaf cell culture according to claim 9, which is characterized in that the culture item
Part control are as follows: cultivation temperature is 15-40 DEG C, intensity of illumination 2000-3000Lx, light application time 6-14h/d, drum rotation speed 5-
50rpm, recyclegas flow velocity are 200-1000mL/min.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111635860A (en) * | 2020-06-17 | 2020-09-08 | 山东大学 | A system and culture method for stem cell culture |
| CN111944692A (en) * | 2020-08-26 | 2020-11-17 | 河南大学 | A kind of animal cell hollow fiber culture system and cell culture method thereof |
| WO2021008571A1 (en) | 2019-07-16 | 2021-01-21 | 信达生物制药(苏州)有限公司 | Cell culture method and application thereof based on high-density and continuous inoculation |
| CN112457979A (en) * | 2020-11-19 | 2021-03-09 | 江西迈柯菲生物医药科技有限公司 | Double-cylinder centrifugal large-scale suspension type cell culture device |
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Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS637783A (en) * | 1986-06-30 | 1988-01-13 | Japan Synthetic Rubber Co Ltd | Bonding method for adhesion-dependent cell to hollow fiber |
| US5162225A (en) * | 1989-03-17 | 1992-11-10 | The Dow Chemical Company | Growth of cells in hollow fibers in an agitated vessel |
| CN1290742A (en) * | 1999-11-16 | 2001-04-11 | 中国科学院力学研究所 | Threed-dimensional stress adjustable rotary cell/tissue cultivator and its cultivating method |
| CN1542122A (en) * | 2003-11-07 | 2004-11-03 | 大连理工大学 | Rotary Cell Culture System |
| CN1969037A (en) * | 2004-06-14 | 2007-05-23 | 普罗拜奥根股份公司 | Liquid/gas phase exposure reactor for cell cultivation |
| US20070122904A1 (en) * | 2000-09-29 | 2007-05-31 | Unisearch Limited | Method and apparatus for culturing cells |
| CN1994478A (en) * | 2006-12-21 | 2007-07-11 | 大连理工大学 | Method for amplifying mensenchymal stem cell under three-dimensional dynamic condition |
| CN101238207A (en) * | 2005-06-29 | 2008-08-06 | 雷格内泰克公司 | Rotatable perfusion time-varying electromagnetic force bioreactor and method of use thereof |
| CN102181364A (en) * | 2011-03-11 | 2011-09-14 | 北京航空航天大学 | Device for culturing in vitro cells through single axis rotating and online shearing |
| CN107400633A (en) * | 2017-08-17 | 2017-11-28 | 上海白泽医疗器械有限公司 | Doughnut exchanger and doughnut switch type culture systems |
| US20170349873A1 (en) * | 2016-06-07 | 2017-12-07 | Terumo Bct, Inc. | Coating a Bioreactor |
| CN208829690U (en) * | 2018-07-20 | 2019-05-07 | 中南大学 | Drum Hollow Fiber Reactor for High Density Culture of Plant Cells |
-
2018
- 2018-07-20 CN CN201810802922.2A patent/CN108949558A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS637783A (en) * | 1986-06-30 | 1988-01-13 | Japan Synthetic Rubber Co Ltd | Bonding method for adhesion-dependent cell to hollow fiber |
| US5162225A (en) * | 1989-03-17 | 1992-11-10 | The Dow Chemical Company | Growth of cells in hollow fibers in an agitated vessel |
| CN1290742A (en) * | 1999-11-16 | 2001-04-11 | 中国科学院力学研究所 | Threed-dimensional stress adjustable rotary cell/tissue cultivator and its cultivating method |
| US20070122904A1 (en) * | 2000-09-29 | 2007-05-31 | Unisearch Limited | Method and apparatus for culturing cells |
| CN1542122A (en) * | 2003-11-07 | 2004-11-03 | 大连理工大学 | Rotary Cell Culture System |
| CN1969037A (en) * | 2004-06-14 | 2007-05-23 | 普罗拜奥根股份公司 | Liquid/gas phase exposure reactor for cell cultivation |
| CN101238207A (en) * | 2005-06-29 | 2008-08-06 | 雷格内泰克公司 | Rotatable perfusion time-varying electromagnetic force bioreactor and method of use thereof |
| CN1994478A (en) * | 2006-12-21 | 2007-07-11 | 大连理工大学 | Method for amplifying mensenchymal stem cell under three-dimensional dynamic condition |
| CN102181364A (en) * | 2011-03-11 | 2011-09-14 | 北京航空航天大学 | Device for culturing in vitro cells through single axis rotating and online shearing |
| US20170349873A1 (en) * | 2016-06-07 | 2017-12-07 | Terumo Bct, Inc. | Coating a Bioreactor |
| CN107400633A (en) * | 2017-08-17 | 2017-11-28 | 上海白泽医疗器械有限公司 | Doughnut exchanger and doughnut switch type culture systems |
| CN208829690U (en) * | 2018-07-20 | 2019-05-07 | 中南大学 | Drum Hollow Fiber Reactor for High Density Culture of Plant Cells |
Non-Patent Citations (4)
| Title |
|---|
| KHAPARDE, SS等: "Effect of Low Culture Temperature on Urokinase Production in Hollow Fiber Reactor", APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, vol. 168, no. 63, pages 1655 - 1663, XP035141030, DOI: 10.1007/s12010-012-9886-2 * |
| 宋克东等: "中空纤维膜在旋转壁式生物反应器内作为细胞载体的性能", 高校化学工程学报, vol. 21, no. 3, pages 488 - 493 * |
| 李峻城等: "中空纤维细胞培养技术", 国外医学.预防.诊断.治疗用生物制品分册, pages 1 - 4 * |
| 王志江等: "动植物细胞培养生物反应器的研究进展", 药物生物技术, vol. 12, no. 12, pages 117 - 120 * |
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|---|---|---|---|---|
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| US12319900B2 (en) * | 2020-01-31 | 2025-06-03 | Rheinisch-Westfälische Technische Hochschule (Rwth) Aachen | Integral gas-introduction and stirring unit for gas-liquid reactors |
| CN111635860A (en) * | 2020-06-17 | 2020-09-08 | 山东大学 | A system and culture method for stem cell culture |
| CN111944692A (en) * | 2020-08-26 | 2020-11-17 | 河南大学 | A kind of animal cell hollow fiber culture system and cell culture method thereof |
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