CN106434341A - Living cell separating system and separating method adopting same - Google Patents
Living cell separating system and separating method adopting same Download PDFInfo
- Publication number
- CN106434341A CN106434341A CN201610844097.3A CN201610844097A CN106434341A CN 106434341 A CN106434341 A CN 106434341A CN 201610844097 A CN201610844097 A CN 201610844097A CN 106434341 A CN106434341 A CN 106434341A
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- container
- fluid pump
- tangential flow
- doughnut
- living cells
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- 238000000034 method Methods 0.000 title claims abstract description 6
- 239000012530 fluid Substances 0.000 claims abstract description 47
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 239000012528 membrane Substances 0.000 claims abstract description 20
- 239000011550 stock solution Substances 0.000 claims abstract description 20
- 238000004113 cell culture Methods 0.000 claims abstract description 17
- 235000012489 doughnuts Nutrition 0.000 claims description 26
- 230000014759 maintenance of location Effects 0.000 claims description 11
- 238000000926 separation method Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000002699 waste material Substances 0.000 claims description 4
- 238000009295 crossflow filtration Methods 0.000 claims description 3
- 239000012510 hollow fiber Substances 0.000 abstract 5
- 230000000694 effects Effects 0.000 description 7
- 238000001914 filtration Methods 0.000 description 3
- 238000002955 isolation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
-
- C—CHEMISTRY; METALLURGY
- 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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Cell Biology (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to a living cell separating system and a separating method adopting the living cell separating system. The separating system comprises a reactor, a hollow fiber tangential flow filter, a pressurizer, a first container, a second container, a first fluid pump and a second fluid pump, wherein the reactor communicates with the hollow fiber tangential flow filter through the pressurizer, the hollow fiber tangential flow filter communicates with the first container, the first container communicates with the second container, a pressure push type isolating membrane is mounted between the first container and the second container, and the first fluid pump and the second fluid pump are connected to the second container. The separating method comprises the following steps: enabling the first fluid pump to pump liquid for the second container, and enabling culture stock solution in the reactor to enter the first container through the hollow fiber tangential flow filter; enabling the second fluid pump to pressure the second container, and enabling the culture stock solution in the first container to flow back to the reactor through the hollow fiber tangential flow filter; and repeating the steps (1) and (2). The stability of the system provided by the invention is easy to control, the risk that the cell culture stock solution is polluted is reduced, and the separating efficiency of the cells is improved.
Description
Technical field
The invention belongs to the detached technical field of biological cell, more particularly to a kind of living cells piece-rate system and its separation
Method.
Background technology
During bio-pharmaceuticals, the ingredient in cell be with high concentration in less, the most economical life of relative scale
Cultivated in thing reactor.Now, the perfusion system for being used must be with cell retention function, to cut as much as possible
Stay the useful medium in bioreactor.Preferably cell retention system should both have good perfusion ability, have again
There is very high cell retention ability, during cell retention, should also not affect the activity of cell as much as possible, can be in most short stop
In time, the cell of retention is fed back in cultivation reactor.
Generally there is the disadvantages such as stability of control system is not high, cell culture stock solution is contaminated in existing living cells piece-rate system
End, and the inefficiency of cell separation, reduce the output of cell pharmaceutical product.
Content of the invention
The technical problem to be solved is to provide a kind of living cells piece-rate system and its separation method, system stability
Property easy-to-use control, reduce the contaminated risk of cell culture stock solution, improve the separation efficiency of cell.
The technical solution adopted for the present invention to solve the technical problems is to provide a kind of living cells piece-rate system, including reaction
Device, doughnut tangential flow filter, supercharger, the first container, second container, first fluid pump and second fluid pump, described
Reactor is connected with the water inlet of doughnut tangential flow filter one end by supercharger, the doughnut tangential flow filtration
The water inlet of the device other end and the first reservoir, are through structure between first container and second container, and first holds
Fluid pressure driving isolating membrane is installed between device and second container, the first fluid pump and second fluid pump pass through pipeline
It is connected to second container.
A kind of preferred embodiment as the present invention, the inside of the doughnut tangential flow filter is cut by cell
Fibrous membrane is stayed to form cell culture stock solution circulation passage, the cavity on the outside of the cell retention fibrous membrane is provided with the discharge of some waste liquids
Mouthful.
Used as another preferred embodiment of the present invention, the doughnut tangential flow filter vertically sets
Put, first container is located at the surface of second container, the isolating membrane is horizontally disposed in the first container and second
Between container.
Used as another preferred embodiment of the present invention, the first fluid pump and second fluid pump pass through common pipe
Road is connected with second container.
It is to provide a kind of to separate using above-mentioned living cells that the present invention solves another technical scheme that its technical problem adopted
System carries out the detached method of living cells, comprises the following steps：
(1) drawing liquid is carried out to second container by first fluid pump, isolating membrane is compressed to second container, the first container is formed
Negative pressure, the cell culture stock solution in reactor is entered in the first container by doughnut tangential flow filter；
(2) terminate drawing liquid, by second fluid pump, second container is pressurizeed, isolating membrane is compressed to the first container, the
Cell culture stock solution inside one container is back to by doughnut tangential flow filter and in the presence of supercharger reaction
In device；
(3) repeat step (1) and (2), making cell culture stock solution iterate through doughnut tangential flow filter is carried out carefully
Born of the same parents' retention is separated.
A kind of preferred embodiment as the present invention, the first fluid pump carries out the time control of drawing liquid to second container
8～12s is made as, the time control pressurizeed to second container by the second fluid pump is 2～4s.
Used as the improvement further to above-mentioned embodiment, the first fluid pump carries out the time of drawing liquid to second container
10s is controlled to, the time control pressurizeed to second container by the second fluid pump is 3s.
Beneficial effect
In the present invention, the power source of system is provided by fluid pump, can the more pressure of accuracy-control system, time
With the parameters such as flow, so as to realize the stability of system and easily controllable；Using liquid, system is controlled, reduces container
Internal isolation membrane damage, reduces the contaminated risk of cell culture stock solution；By turning between drawing liquid operating mode and pressurization operating mode
The time difference of presence is changed, an instantaneous pressure effect for filtering inside and outside instantaneous differential pressure, then coordinating supercharger is provided, is improve thin
Born of the same parents retain detached effect, so as to be conducive to improve production efficiency.
Description of the drawings
The system equipment attachment structure figure of Fig. 1 present invention.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limit the scope of the present invention.In addition, it is to be understood that after the content for having read present invention instruction, people in the art
Member can be made various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
A kind of living cells piece-rate system as shown in Figure 1, including reactor, doughnut tangential flow filter, supercharger,
First container, second container, first fluid pump and second fluid pump.
Doughnut tangential flow filter is vertically arranged, and reactor is tangentially flow through with doughnut by supercharger
The water inlet connection of filter upper end, the water inlet of doughnut tangential flow filter lower end and the first reservoir.Doughnut
The inside of tangential flow filter forms cell culture stock solution circulation passage by cell retention fibrous membrane, and cell culture stock solution is in warp
Carry out cell while crossing the circulation passage to dam separately, the cavity on the outside of cell retention fibrous membrane is provided with the discharge of some waste liquids
Mouthful, the waste liquid for filtering is discharged outside doughnut tangential flow filter.
First container is located at the surface of second container, is through structure between the first container and second container, and first
Fluid pressure driving isolating membrane is horizontally installed between container and second container.First fluid pump and second fluid pump are by pipe
Road is connected to second container, and first fluid pump is connected with second container by common pipeline with second fluid pump, reduces pipeline
Laying.
One kind is provided below carries out the detached method of living cells using above-mentioned living cells piece-rate system, comprises the following steps：
(1) drawing liquid operating mode：Start first fluid pump, drawing liquid, drawing liquid time are carried out by first fluid pump to second container
8～12s is controlled to, is advisable with being set to 10s.Isolating membrane is compressed to second container in the presence of second container negative pressure, so as to
So that the first container forms negative pressure, the cell culture stock solution in reactor enters into first by doughnut tangential flow filter
In container, now supercharger is opened into atmospheric pressure state.
(2) pressurization operating mode：Terminate drawing liquid, stop first fluid pump, start second fluid pump, by second fluid pump to the
Two containers are pressurizeed, and pressing time is controlled to 2～4s, are advisable with being set to 3s.Isolating membrane is in the effect of second container malleation
Lower to the first container compress, the cell culture stock solution the first container inside by doughnut tangential flow filter and be pressurized
It is back in the presence of device in reactor, now supercharger work so that doughnut tangential flow filter is to supercharger end shape
Become certain pressure.
(3) by first fluid pump and second fluid pump mutual switch operating repeat step (1) and step (2), cell is made
Culture stock solution iterates through doughnut tangential flow filter, and under the pressurization of conduit pressurizer, realizes cell and dam
Function is so as to reach detached effect.
The system provides the power source of system by fluid pump, can the more pressure of accuracy-control system, time and stream
The parameters such as amount, so as to realize the stability of system and easily controllable.Using liquid, system is controlled, reduces inside container
Isolation membrane damage, reduce the contaminated risk of cell culture stock solution.Deposited by changing between drawing liquid operating mode and pressurization operating mode
Time difference, an instantaneous pressure effect for filtering inside and outside instantaneous differential pressure, then coordinating supercharger is provided, is improve cell and cut
Detached effect is stayed, so as to be conducive to improve production efficiency.
Claims (7)
1. a kind of living cells piece-rate system, including reactor, doughnut tangential flow filter, supercharger, the first container, second
Container, first fluid pump and second fluid pump, it is characterised in that：The reactor is tangentially flow through with doughnut by supercharger
The water inlet connection of filter one end, the water inlet of the doughnut tangential flow filter other end and the first reservoir, institute
It is through structure to state between the first container and second container, is provided with Fluid pressure promotion between the first container and second container
Formula isolating membrane, the first fluid pump and second fluid pump are connected to second container by pipeline.
2. a kind of living cells piece-rate system according to claim 1, it is characterised in that：The doughnut tangential flow filtration
The inside of device forms cell culture stock solution circulation passage, the chamber on the outside of the cell retention fibrous membrane by cell retention fibrous membrane
Body is provided with some waste liquid outlets.
3. a kind of living cells piece-rate system according to claim 1, it is characterised in that：The doughnut tangential flow filtration
Device is vertically arranged, first container be located at second container surface, the isolating membrane horizontally disposed in
Between first container and second container.
4. a kind of living cells piece-rate system according to claim 1, it is characterised in that：The first fluid pump and second
Body pump is connected with second container by common pipeline.
5. the living cells piece-rate system described in a kind of utilization claim 1 carries out the detached method of living cells, comprises the following steps：
(1) drawing liquid is carried out to second container by first fluid pump, isolating membrane is compressed to second container, and the first container forms negative
Pressure, the cell culture stock solution in reactor is entered in the first container by doughnut tangential flow filter；
(2) terminate drawing liquid, by second fluid pump, second container is pressurizeed, isolating membrane is compressed to the first container, first holds
Cell culture stock solution inside device by doughnut tangential flow filter and is back to reactor in the presence of supercharger
In；
(3) repeat step (1) and (2), make cell culture stock solution iterate through doughnut tangential flow filter and carry out cell and cut
Stay and separate.
6. a kind of living cells separation method according to claim 5, it is characterised in that：The first fluid pump holds to second
Device carry out drawing liquid time control be 8～12s, the time control pressurizeed to second container by the second fluid pump be 2～
4s.
7. a kind of living cells separation method according to claim 6, it is characterised in that：The first fluid pump holds to second
It is 10s that device carries out the time control of drawing liquid, and the time control pressurizeed to second container by the second fluid pump is 3s.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610844097.3A CN106434341A (en) | 2016-09-22 | 2016-09-22 | Living cell separating system and separating method adopting same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610844097.3A CN106434341A (en) | 2016-09-22 | 2016-09-22 | Living cell separating system and separating method adopting same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106434341A true CN106434341A (en) | 2017-02-22 |
Family
ID=58166882
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610844097.3A Pending CN106434341A (en) | 2016-09-22 | 2016-09-22 | Living cell separating system and separating method adopting same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106434341A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111454902A (en) * | 2020-04-08 | 2020-07-28 | 郭华 | Method and device for separating and enriching white blood cells, stem cells and/or pathogens from blood |
| CN114874911A (en) * | 2022-06-07 | 2022-08-09 | 上海拜高乐生物技术有限公司 | Cell and excrement separator thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6544424B1 (en) * | 1999-12-03 | 2003-04-08 | Refined Technology Company | Fluid filtration system |
| CN103298538A (en) * | 2010-08-25 | 2013-09-11 | J·谢瓦利茨 | Fluid filtration systems |
| US20160222337A1 (en) * | 2012-10-02 | 2016-08-04 | David Serway | Method for Proliferation of Cells Within a Bioreactor Using a Disposable Pumphead and Filter Assembly |
| CN206109422U (en) * | 2016-09-22 | 2017-04-19 | 上海元创生物科技有限公司 | Living cells piece -rate system |
-
2016
- 2016-09-22 CN CN201610844097.3A patent/CN106434341A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6544424B1 (en) * | 1999-12-03 | 2003-04-08 | Refined Technology Company | Fluid filtration system |
| CN103298538A (en) * | 2010-08-25 | 2013-09-11 | J·谢瓦利茨 | Fluid filtration systems |
| US20160222337A1 (en) * | 2012-10-02 | 2016-08-04 | David Serway | Method for Proliferation of Cells Within a Bioreactor Using a Disposable Pumphead and Filter Assembly |
| CN206109422U (en) * | 2016-09-22 | 2017-04-19 | 上海元创生物科技有限公司 | Living cells piece -rate system |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111454902A (en) * | 2020-04-08 | 2020-07-28 | 郭华 | Method and device for separating and enriching white blood cells, stem cells and/or pathogens from blood |
| CN114874911A (en) * | 2022-06-07 | 2022-08-09 | 上海拜高乐生物技术有限公司 | Cell and excrement separator thereof |
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170222 |