CN104415405B - A kind of stirring type bioreactor of the convertible flow direction being applied to artificial liver - Google Patents
A kind of stirring type bioreactor of the convertible flow direction being applied to artificial liver Download PDFInfo
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- CN104415405B CN104415405B CN201310376469.0A CN201310376469A CN104415405B CN 104415405 B CN104415405 B CN 104415405B CN 201310376469 A CN201310376469 A CN 201310376469A CN 104415405 B CN104415405 B CN 104415405B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/14—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
- A61M1/16—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis with membranes
- A61M1/1698—Blood oxygenators with or without heat-exchangers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/14—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
- A61M1/16—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis with membranes
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- Urology & Nephrology (AREA)
- Heart & Thoracic Surgery (AREA)
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- Engineering & Computer Science (AREA)
- Vascular Medicine (AREA)
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- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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- External Artificial Organs (AREA)
Abstract
The invention discloses the stirring type bioreactor of a kind of convertible flow direction being applied to artificial liver, comprise the bioreactor of band stirring paddle, load in reactor and be embedded with hepatocellular microcapsule.Owing to microcapsule proportion is close with water, under stirring, it is readily formed suspension, it is thus possible under the continuous perfusion operating condition of high flow velocities, realize microcapsule to be sufficiently mixed with liquid, simultaneously facilitate solid-liquid mass transfer, it is ensured that the nutrient substance supply that in microcapsule, hepatocyte is sufficient discharges with the efficient of active substance.Additionally, the microcapsule being likely to occur in order to avoid long-time Perfused or the situation of clogged with fragments filter screen, reactor entrance end is respectively equipped with electromagnetic valve and peristaltic pump, can realize flowing to conversion, reach the effect of recoil, it is to avoid pipeline blockage, keep solid-liquid mix homogeneously in reactor.The present invention can realize good solid-liquid mixing and mass exchange under the conditions of bigger solid-to-liquid ratio, higher perfusion flow velocity and higher cell density, is prevented effectively from perfusion dead space and invalid perfusion, plays the therapeutical effect of artificial liver to greatest extent.
Description
Technical field
The present invention relates to a kind of armarium, the Biotype artificial liver of a kind of clinical treatment acute hepatic failure or
The core apparatus of hybrid artificial liver support system: the stirring type bioreactor of the convertible flow direction.
Background technology
Acute hepatic failure is common critical illness, and mortality rate is up to 65～95%.Artificial liver in treatment has become treatment at present
The important means of acute hepatic failure.Artificial liver support system, is external machinery or physics and chemistry device, temporarily auxiliary or completely generation
For the function of exhaustion liver, remove various harmful substance, the metabolic function of compensatory liver.And it is mounted with the hepatocellular life of high activity
Thing type artificial liver, can perform the repertoires such as liver synthesis, conversion in theory, be trend and the trend of artificial liver future development.
Bioreactor is the core component of Biotype artificial liver, as the place of hepatocyte with plasma substance exchange, it is desirable to there is high pass
The mass exchange of amount, to have enough hepatocyte quantity simultaneously.
Applying most hollow-fiber bioreactors at present, its advantage is that specific surface area is big, it is simple to the transhipment of metabolite,
And reactor dead space is less, but due to hepatocyte skewness in the reactor, also easily cause cell viability decline, anoxia and thing
The inefficient problem of mass transter, and reactor is difficult to amplify, it is more difficult to meet cell concentration required during treatment.In recent years, based on
The Biotype artificial liver device of microencapsulated hepatocyte gets the attention.Microcapsule technology is by having half by cell embedding
In infiltrative polymeric film, hepatocyte can be protected from immune attack, reduce and shear injury, can guarantee that again good simultaneously
Mass exchange.Additionally, microcapsule has bigger mass transfer specific surface, moreover it is possible to provide the intracellular loops of three-dimensional class loading to cell
Border.The Biotype artificial liver reactor assembly built based on microencapsulated hepatocyte, loads conveniently, easily amplifies, and is improving thing
Mass transter and cytoactive aspect have the advantage of uniqueness, can play the therapeutical effect of Biotype artificial liver more efficiently.
It is currently based on the Biotype artificial liver reactor of microencapsulated hepatocyte and mainly has microcapsule-fixed bed reactors and micro-
Two kinds of forms of capsule-suspension reactor.Microcapsule-fixed bed reactors have higher loading cells density, and can reach higher
Operation flow velocity, it is possible to meet the oxygen supply of cell, it is achieved good mass exchange, but microcapsule presser sensor, high flow rate
The high post pressure that perfusion produces is easily caused microcapsule deformation and blocks bed, and high shear force and lasting column pressure easily cause micro-simultaneously
Capsule crushes.Microcapsule-suspension reactor principal mode is fluidized-bed reactor, and its advantage is that mass transfer is uniform, and shearing force is little, post pressure
Low, it is not easily formed intrinsic passage and perfusion dead space, but owing to Microcapsules Size is little, proportion is close with blood plasma, easy during fluidisation
Being taken out of by liquid, the operation flow velocity that can reach is relatively low, limits mass exchange, also cannot meet hepatocellular oxygen demand simultaneously
Amount.It addition, form bigger voidage during fluidization operation, the plasma volume of cardiopulmonary bypass in patients will certainly be increased, increase patient and bear
Load, hepatocellular density in simultaneously reducing reactor.
It is thus desirable to develop the bioreactor of novel applicable microencapsulated hepatocyte system, there is high loading cells density
Less reactor design volume；Can be real under the continuous perfusion of the blood plasma of high flow velocities and bigger solid-to-liquid ratio operating condition
Existing microcapsule and blood plasma are sufficiently mixed, cell is good with blood plasma (blood) mass exchange, provide hepatocyte sufficient nutrient
Matter (comprises oxygen), and the functional molecular of hepatocytes secrete can timely enter blood plasma (blood) and play a role, and can be longer
Time keeps cell growth activity and protects cells from shearing and immunologic cytotoxicity etc..
Summary of the invention
Therefore, the stirring type bioreactor of a kind of convertible flow direction being applied to artificial liver of present invention design, utilize
Microcapsule proportion is close with water, is easily formed this feature of suspension, utilizes churned mechanically immixture, it is possible in higher filling
Under stream flow velocity and bigger solid-to-liquid ratio operating condition, it is achieved microcapsule is sufficiently mixed with blood plasma, uses the convertible flow direction simultaneously
Design, pipeline can be recoiled, it is to avoid pipeline blockage, microcapsule and plasma uniform admixture can be maintained for a long time.
The technical solution used in the present invention is:
The stirring type bioreactor of a kind of convertible flow direction being applied to artificial liver, including main body reactor and stirring
Oar, main body reactor is a hermetic container, and stirring paddle is arranged in main body reactor, is provided with membrane type oxygen in main body reactor
Clutch；The import of membrane oxygenator is connected with oxygen source, the outlet emptying of membrane oxygenator；In the top of main body reactor or
Top is provided with a port of liquid entrance, liquid entrance and the first two-position three way magnetic valve and is connected, the first two-position three way
Second port of electromagnetic valve is connected with wet tank through the first peristaltic pump, the 3rd port warp of the first two-position three way magnetic valve
Second peristaltic pump) it is connected with liquid collection storage tank；Be provided with liquid discharge entrance in the bottom of main body reactor or bottom, liquid comes in and goes out
Mouth is connected with a port of the second two-position three way magnetic valve, and second port of the second two-position three way magnetic valve is wriggled through first
Pump is connected with wet tank, and the 3rd port of the second two-position three way magnetic valve collects storage tank phase through the second peristaltic pump with liquid
Even.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that:
Microencapsulated hepatocyte and solution is loaded in main body reactor, under the effect of stirring paddle, can be at continuous perfusion
Realize microcapsule under state to mix with the uniform of solution；Reactor two ends be respectively provided with first, second two-position three way magnetic valve and
First, second peristaltic pump, it is possible to achieve flow to conversion, reaches the effect of recoil, it is to avoid pipeline blockage, keeps solid-liquid in reactor
Mix homogeneously.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that:
Liquid entrance and liquid discharge entrance one end are in main body reactor, and the other end is electric with the first two-position three way respectively
One port of magnet valve and the second two-position three way magnetic valve is connected；
Liquid entrance one end and liquid discharge entrance one end of being positioned at main body reactor are in the molten of main body reactor
Under liquid liquid level；Or, liquid entrance one end and/or liquid discharge entrance one end of being positioned at main body reactor are communicated to by the road
Under liquid level of solution in main body reactor.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that:
The import of membrane oxygenator is connected with the oxygen source outside main body reactor by pipeline, going out of membrane oxygenator
Mouth stretches out the outside emptying of main body reactor by pipeline.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that:
It is provided with liquid discharge porch in the bottom of main body reactor or bottom and is provided with filter screen；
It is provided with at liquid entrance in the top of main body reactor or top and is provided with filter screen；
Described filter screen material is silkworm silk, tinsel or synthetic fibers, its aperture 100～800 mesh.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that: stirring paddle is electricity
The mechanical agitation oar that machine drives；
Described stirring paddle material is politef or stainless steel material, and for monolayer or multiple impellers, blade is
Double blades or many blades.
The stirring type bioreactor of the described convertible flow direction being applied to artificial liver, it is characterised in that: described reaction
Device liquid entrance end and liquid discharge entrance are provided with first, second two-position three way magnetic valve and first, second peristaltic pump, can realize
Manually or automatically flow to conversion.
When reactor uses, stirring paddle stir speed (S.S.) is 10rpm～1000rpm, and it can realize under continuous perfusion operates
Microcapsule uniformly mixes with liquid, its continuous perfusion flow 50ml/min～1000ml/min.
The invention discloses the stirring type bioreactor of a kind of convertible flow direction being applied to artificial liver, comprise band stirring
The bioreactor of oar, loads in reactor and is embedded with hepatocellular microcapsule.Owing to microcapsule proportion is close with water, in stirring
Under effect, it is easy to form suspension, it is thus possible under the continuous perfusion operating condition of high flow velocities, it is achieved microcapsule and liquid
Body is sufficiently mixed, and simultaneously facilitates solid-liquid mass transfer, it is ensured that nutrient substance supply and the active substance that in microcapsule, hepatocyte is sufficient
Efficiently release.Additionally, the microcapsule being likely to occur in order to avoid long-time Perfused or the situation of clogged with fragments filter screen, reaction
Device entrance end is respectively equipped with electromagnetic valve and peristaltic pump, can realize flowing to conversion, reach the effect of recoil, it is to avoid pipeline blockage,
Keep solid-liquid mix homogeneously in reactor.The present invention can be in bigger solid-to-liquid ratio, higher perfusion flow velocity and high hepatocyte density
Under the conditions of realize the mixing of good solid-liquid and mass exchange, be prevented effectively from perfusion dead space and invalid perfusion, play to greatest extent
The therapeutical effect of artificial liver.
Accompanying drawing explanation
Fig. 1 is the structural representation of the present invention, wherein: 1 is main body reactor；2 drive the stirring paddle of motor for band；3 are
Membrane oxygenator；4 is filter screen；5 for being embedded with hepatocellular microcapsule；6,7 is first, second two-position three way magnetic valve；8 is
One peristaltic pump, 9 is the second peristaltic pump.A, a ' it is blood plasma；B is oxygen.
Fig. 2 is that the stirring type bioreactor of the convertible flow direction runs microencapsulated forms after 10 hours continuously: the most left figure
For the microencapsulated forms before running；Right figure is to run the form of microcapsule after 10 hours.
Detailed description of the invention
As it is shown in figure 1, the stirring type bioreactor of a kind of convertible flow direction being applied to artificial liver, react including main body
Device 1 and stirring paddle 2, main body reactor 1 is a hermetic container, and stirring paddle 2 is arranged in main body reactor, in main body reactor
It is provided with membrane oxygenator 3；The import of membrane oxygenator 3 is connected with oxygen source, the outlet emptying of membrane oxygenator 3；In master
The top of reactor according or top are provided with a port of liquid entrance, liquid entrance and the first two-position three way magnetic valve 6
Being connected, second port of the first two-position three way magnetic valve 6 is connected with wet tank through the first peristaltic pump 8, the first two-position three way
3rd port of electromagnetic valve 6 is collected storage tank through the second peristaltic pump 9 with liquid and is connected；Set in the bottom of main body reactor or bottom
The port having liquid discharge entrance, liquid discharge entrance and the second two-position three way magnetic valve 7 is connected, the second two-position three way magnetic valve
Second port of 7 is connected with wet tank through the first peristaltic pump 8, and the 3rd port of the second two-position three way magnetic valve 7 is through
Two peristaltic pumps 9 are collected storage tank with liquid and are connected.It is provided with liquid discharge porch in the bottom of main body reactor or bottom and is provided with filter screen
4；
It is provided with at liquid entrance in the top of main body reactor or top and is provided with filter screen 4；Load in main body reactor
Microencapsulated hepatocyte 5 and solution, under the immixture of stirring paddle, can operate at higher perfusion flow velocity and bigger solid-to-liquid ratio
Under the conditions of, it is achieved microcapsule is sufficiently mixed with blood plasma, uses the design of the convertible flow direction simultaneously, can recoil pipeline,
Avoid pipeline blockage, microcapsule and plasma uniform admixture can be maintained for a long time.
Described stirring paddle is motor-driven mechanical agitation oar, is in main body inside reactor, and stirring paddle lowermost end leans on
Nearly main body reactor bottom；Its material is politef or stainless steel material, and for monolayer or multiple impellers, blade is double
Blade or many blades.
Embodiment
Carry hepatocyte microcapsule to prepare: hepatocyte and mass concentration 1.5% sodium alginate suspendible, cell density 1 × 107～1
×108Cells/ml, application high-pressure electrostatic preparing instrument instills the CaCl of mass concentration 1.1%2In solution, the calcium alginate plastic of formation
Pearl with 0.05% polylysin solution react film forming.
Prepare without cell microcapsule: method is as before, 1.5% sodium alginate, and application high-pressure electrostatic preparing instrument instills 1.1%
CaCl2In solution, the calcium alginate plastic beads of formation reacts film forming with the polylysin solution of 0.05%.
Main body reactor: for the glass reactor of cylindricality, reactor inner chamber is, reactor bottom
For stainless steel filtering net, aperture is 200 mesh.
Stirring paddle: for the upper stirring of motor-driven politef material, can be double in 0～1200rpm rotational speed regulation
Leaf, oar width 40mm.
The stirring type bioreactor of the convertible flow direction runs: particle diameter is 200 μm～400 μm microcapsules, and packing volume is
80ml, packed height 40mm, solid-to-liquid ratio 2:1, continuous perfusion flow 200ml/min, when stir speed (S.S.) reaches 200rpm, microcarrier
It is evenly distributed, does not occurs in bottom piling up.After continuous pouring 10h, microencapsulated forms keeps good, percentage of head rice be maintained at 99% with
Upper (such as Fig. 2).
Flow to conversion: as Fig. 1, reactor entrance end and gateway end respectively set first, second two-position three way magnetic valve 6,7
With first, second peristaltic pump 8,9, under normal perfusion state, the first two-position three way three-way magnetic valve 6 left side path is opened, right
Limit path Guan Bi, and the second two-position three way three-way magnetic valve 7 left side path Guan Bi, the unlatching of the right path, liquid flows to such as a institute
Show, when needs conversion flows to, either manually or automatically give the three-way magnetic valve signal of telecommunication, then the first two-position three way threeway electromagnetism
Valve 6 left side path Guan Bi, the right path is opened, and the second two-position three way three-way magnetic valve 7 left side path is opened, and the right path closes
Closing, liquid flows to as shown in a ', and liquid flows to conversion, reaches the purpose of recoil.
Claims (8)
1. it is applied to a stirring type bioreactor for the convertible flow direction of artificial liver, including main body reactor (1) and stirring
Oar (2), main body reactor (1) is a hermetic container, and stirring paddle (2) is arranged in main body reactor, sets in main body reactor
It is equipped with membrane oxygenator (3)；The import of membrane oxygenator (3) is connected with oxygen source, the outlet emptying of membrane oxygenator (3)；
The of liquid entrance, liquid entrance and the first two-position three way magnetic valve (6) it is provided with in the top of main body reactor or top
One port is connected, and second port of the first two-position three way magnetic valve (6) is connected with wet tank through the first peristaltic pump (8),
3rd port of the first two-position three way magnetic valve (6) is collected storage tank through the second peristaltic pump (9) with liquid and is connected；React in main body
The bottom of device or bottom are provided with first port phase of liquid discharge entrance, liquid discharge entrance and the second two-position three way magnetic valve (7)
Even, second port of the second two-position three way magnetic valve (7) is connected with wet tank through the first peristaltic pump (8), the two or two three
3rd port of three-way electromagnetic valve (7) is collected storage tank through the second peristaltic pump (9) with liquid and is connected.
2. according to the stirring type bioreactor of the convertible flow direction being applied to artificial liver described in claim 1, it is characterised in that:
Microencapsulated hepatocyte (5) and solution is loaded, under the effect of stirring paddle, it is possible at continuous perfusion shape in main body reactor
Realize microencapsulated hepatocyte (5) under state to mix with the uniform of solution；Main body reactor two ends are respectively provided with first, second 2 three
Three-way electromagnetic valve (6,7), it is possible to realize flowing to conversion, reach the effect of recoil, it is to avoid pipeline blockage, in keeping reactor, solid-liquid mixes
Close uniformly.
3. according to the stirring type bioreactor of the convertible flow direction being applied to artificial liver described in claim 1, it is characterised in that:
Liquid entrance and liquid discharge entrance one end are in main body reactor, the other end respectively with the first two-position three way magnetic valve
(6) port of with the second two-position three way magnetic valve (7) is connected；
It is positioned at liquid entrance one end of main body reactor and liquid discharge entrance one end is in the solution liquid of main body reactor
Under face；Or, liquid entrance one end and/or liquid discharge entrance one end of being positioned at main body reactor are communicated to main body by the road
Under liquid level of solution in reactor.
4. according to the stirring type bioreactor of the convertible flow direction being applied to artificial liver described in claim 1, it is characterised in that:
The import of membrane oxygenator (3) is connected by the oxygen source that pipeline is outside with main body reactor (1), membrane oxygenator
(3) outlet stretches out the outside emptying of main body reactor (1) by pipeline.
5. according to the stirring type bioreactor of the convertible flow direction being applied to artificial liver described in claim 1, it is characterised in that:
It is provided with liquid discharge porch in the bottom of main body reactor or bottom and is provided with filter screen (4)；
It is provided with at liquid entrance in the top of main body reactor or top and is provided with filter screen (4)；
Described filter screen material is silkworm silk, tinsel or synthetic fibers, and its aperture is 100～800 mesh.
6. according to the stirring type bioreactor of the convertible flow direction being applied to artificial liver described in claim 1, it is characterised in that:
Stirring paddle is motor-driven mechanical agitation oar；
Described stirring paddle material is politef or stainless steel material, and for monolayer or multiple impellers, blade is double oars
Leaf or many blades.
7. being applied to the stirring type bioreactor of the convertible flow direction of artificial liver described in claim 1 or 2, its feature exists
In: described liquid reactor is imported and exported and liquid discharge entrance is respectively equipped with first, second two-position three way magnetic valve (6,7), energy
Realize manually or automatically flowing to conversion.
8. being applied to the stirring type bioreactor of the convertible flow direction of artificial liver described in claim 1 or 6, its feature exists
In: when reactor uses, stirring paddle stir speed (S.S.) is 10rpm～1000rpm, and it can realize microencapsulation under continuous perfusion operates
Hepatocyte (5) uniformly mixes with liquid, and its continuous perfusion flow is 50ml/min～1000ml/min.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310376469.0A CN104415405B (en) | 2013-08-26 | 2013-08-26 | A kind of stirring type bioreactor of the convertible flow direction being applied to artificial liver |
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| CN201310376469.0A CN104415405B (en) | 2013-08-26 | 2013-08-26 | A kind of stirring type bioreactor of the convertible flow direction being applied to artificial liver |
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| Publication Number | Publication Date |
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| CN104415405A CN104415405A (en) | 2015-03-18 |
| CN104415405B true CN104415405B (en) | 2016-12-28 |
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| CN113892483A (en) * | 2021-09-07 | 2022-01-07 | 奇点医疗科技(广州)有限公司 | Anti-layering filling structure and filling device |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4324662A (en) * | 1979-12-28 | 1982-04-13 | Baxter Travenol Laboratories, Inc. | Flow reversal in a dialyzer |
| GB2276566A (en) * | 1993-03-24 | 1994-10-05 | Saitekku Kabushiki Kaisha | Haemodialyser unit |
| US6308737B1 (en) * | 2000-03-10 | 2001-10-30 | Transonic Systems, Inc. | Method and apparatus for selectively reversing flow between a dialyzer and a patient access |
| CN101711898B (en) * | 2009-12-14 | 2012-05-23 | 浙江大学 | Microcapsule suspension stirring pool type bioreactor for artificial liver |
| CN201643113U (en) * | 2010-03-19 | 2010-11-24 | 南方医科大学珠江医院 | Control system for two-way perfusion type bioreactor |
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