CN106497783B - A kind of dynamic circulation cell culture apparatus and cell culture processes - Google Patents
A kind of dynamic circulation cell culture apparatus and cell culture processes Download PDFInfo
- Publication number
- CN106497783B CN106497783B CN201610800907.5A CN201610800907A CN106497783B CN 106497783 B CN106497783 B CN 106497783B CN 201610800907 A CN201610800907 A CN 201610800907A CN 106497783 B CN106497783 B CN 106497783B
- Authority
- CN
- China
- Prior art keywords
- culture solution
- cell
- delivery pump
- culture
- porcine hgf
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/10—Hollow fibers or tubes
-
- 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
- C12M29/12—Pulsatile flow
-
- 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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/20—Degassing; Venting; Bubble traps
-
- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/38—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of metabolites or enzymes in the cells
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Abstract
The present invention relates to technical field of cell culture more particularly to a kind of dynamic circulation cell culture apparatus and cell culture processes.Porcine HGF can be sporadically supplemented for the cell growth chamber, improve cell culture efficiency.A kind of dynamic circulation cell culture apparatus, comprising: hollow fiber reactor, the hollow fiber reactor include culture solution channel and cell growth chamber, Porcine HGF delivery pump；The input terminal of the Porcine HGF delivery pump is for inputting Porcine HGF, the inlet communication of the output end of the Porcine HGF delivery pump and the cell growth chamber, the Porcine HGF delivery pump are used to convey Porcine HGF to the cell growth chamber.
Description
Technical field
The present invention relates to technical field of cell culture more particularly to a kind of dynamic circulation cell culture apparatus and cell culture sides
Method.
Background technique
T cell is broken up by intrathymic lymphoid stem cell, is that quantity is most in lymphocyte, function is most complicated
A kind of cell.It can be assisted with direct killing target cell or B cell is inhibited to generate antibody, have silk point to specific antigen and rush
Split former responsing reaction and generation Porcine HGF etc., chest of the mature T cell through blood distribution to peripheral immune organ
Gland relies on area and settles down, and can be recycled through lymphatic vessel, peripheral blood and tissue fluid etc., and cellular immunity and immunological regulation etc. are played
Function.
Evolution in relation to T cell and the research of it and cancer achieve many progress in recent decades.Especially 21
The life Noah's ark plan that the beginning of the century mankind start for the evolution of T cell and the research of it and cancer be even more achieve it is breakthrough
Progress.Human body can naturally-produced T cell, but negligible amounts.Successful incubation T cell, and this cell is largely injected into patient
In vivo, it is possibly realized with enhancing immune system.
There are two main classes for existing cell culture processes: first kind method is high-grade laminar flow using Tissue Culture Dish
Cover and carbon dioxide incubator carry out cell culture, and such methods efficiency is lower, is not able to satisfy clinical demand；Second class method is
Cultivated the cells by hollow fiber reactor, referring to Fig. 1, the hollow fiber reactor 1 include hollow fiber conduit with
And the cylindrical shell (not shown) of the installation hollow fiber conduit, the inside of the hollow fiber conduit is for inputting culture
Liquid (in figure shown in arrow a) is culture solution channel 11, and the gap between the outside of the hollow fiber conduit and the shell is used for
Inoculating cell is cell growth chamber 12, by the way that culture solution is flowed through the hollow fiber conduit, and passes through the hollow fiber conduit
Aperture enters the cell growth chamber 12 for cells with nutrient, in the cell cultivation process in the cell growth chamber 12 constantly
It generates waste liquid and drains into outside, constantly provide culture solution in this way for cell culture, the dosage of culture solution is larger, and will give up
The Porcine HGF for being conducive to cell growth that generated in cell cultivation process can be discharged together when being discharged for liquid, cell culture
Environment is unstable, is unfavorable for cell growth.
Summary of the invention
The embodiment of the present invention provides a kind of dynamic circulation cell culture apparatus and cell culture processes, can be the cell
Growth chamber sporadically supplements Porcine HGF, improves cell culture efficiency.
In order to achieve the above objectives, the embodiment of the present invention adopts the following technical scheme that
On the one hand, the embodiment of the present invention provides a kind of dynamic circulation cell culture apparatus, comprising:
Hollow fiber reactor, the hollow fiber reactor include culture solution channel and cell growth chamber；
Porcine HGF delivery pump；The input terminal of the Porcine HGF delivery pump is for inputting cell growth factor
Son, the inlet communication of the output end of the Porcine HGF delivery pump and the cell growth chamber, the Porcine HGF
Delivery pump is used to convey Porcine HGF to the cell growth chamber.
Preferably, the incubator further includes Porcine HGF storage tank, the outlet of the Porcine HGF storage tank with
The input terminal of the cell factor delivery pump is connected to, and the bottom of the Porcine HGF storage tank is provided with import, the cell
The entrance of growth factor storage tank is provided with shut-off valve, and the shut-off valve is used for when closed by the Porcine HGF storage tank
Interior Porcine HGF is isolated from the outside.
Preferably, the incubator further includes pressure extension box and culture solution delivery pump, in the pressure extension box filled with
Gas, the bottom of the pressure extension box are connected to the culture solution channel or the cell growth chamber；
The input terminal of the culture solution delivery pump connects for inputting culture solution, the import in output end and the culture solution channel
Logical, culture solution for being flowed through the culture solution channel by the culture solution delivery pump in the form of impulse wave, so that the culture
The pressure difference of liquid channel and the cell growth chamber is positive and negative alternately to be changed, and the pressure extension box is used for the change by gas volume
Change the pressure balance so that the culture solution channel and the cell growth chamber.
Preferably, the dynamic circulation cell culture apparatus further includes waste tank, waste liquid delivery pump and the first sampler, institute
The input terminal of waste liquid delivery pump and the outlet in the culture solution channel are stated, output end is connected to the waste tank, described
The outlet of the collection terminal of first sampler and the culture solution channel, the first sampler are used for logical to the culture solution is flowed through
The culture solution in road is sampled.
It is further preferred that the dynamic circulation cell culture apparatus further includes the second sampler, second sampler
Collection terminal is connected to the cell growth chamber, and second sampler is used for the cell and culture solution in the cell growth chamber
It is sampled.
Optionally, the dynamic circulation cell culture apparatus further includes cellular collector, the import of the cellular collector with
Pipeline connection between the Porcine HGF storage tank and the Porcine HGF delivery pump, the cellular collector are used for
The cell collection in the cell growth chamber is got up in the Porcine HGF delivery pump antiport.
It is further preferred that the dynamic circulation cell culture apparatus further includes controller, the controller respectively with it is described
Shut-off valve, culture solution delivery pump, Porcine HGF delivery pump, the electrical connection of waste liquid delivery pump, for controlling the shut-off valve, training
The opening and closing of nutrient solution delivery pump, Porcine HGF delivery pump and waste liquid delivery pump, and control the culture solution delivery pump
Culture solution is flowed through to the culture solution channel in the form of impulse wave, control the Porcine HGF delivery pump Positive work or
Person's antiport.
On the other hand, the embodiment of the present invention provides a kind of dynamic circulation cell culture processes, is applied to as described above dynamic
State circulating cells incubator, comprising:
Porcine HGF is conveyed to the cell growth chamber by the Porcine HGF delivery pump, so that described thin
The intracavitary Porcine HGF content of intracellular growth is maintained in the first preset range.
Optionally, the method also includes: every the first preset time, by first sampler to flowing through the training
The culture solution in nutrient solution channel is sampled detection, if Metabolites Concentration is more than or equal to the first preset value in the culture solution,
The waste liquid delivery pump then is controlled by the controller to open, and the culture solution for flowing through the culture solution channel is drained into described useless
In liquid storage tank.
Further, the method also includes: every the second preset time, by second sampler to the cell
The cell and culture solution of growth chamber, which are sampled, detects cell density therein, if the cell density is more than or equal to the
Two preset values then control the Porcine HGF delivery pump antiport by the controller, by the cell growth chamber
In cell delivery into the cellular collector.
Optionally, described that the Porcine HGF delivery pump antiport is controlled by the controller, it will be described thin
After cell delivery in intracellular growth chamber is into the cellular collector further include: passing through the controller, to control the cell raw
Long factor delivery pump Positive work, by the cellular collector to the cell growth chamber inoculating cell.
A kind of dynamic circulation cell culture apparatus and cell culture processes provided in an embodiment of the present invention, in cell cultivation process
In, as cell is grown, growth factor can be constantly consumed, while with the continuous discharge of waste liquid, the cell growth chamber
Porcine HGF content can reduce, and by the way that Porcine HGF delivery pump is arranged, pass through the Porcine HGF delivery pump
Porcine HGF sporadically can be conveyed to the cell growth chamber, the Porcine HGF content is kept to be maintained at one
In fixed range, so as to improve cell culture effect.
Detailed description of the invention
Fig. 1 is a kind of structural schematic diagram for cell culture reactor that the prior art provides；
Fig. 2 is a kind of structural schematic diagram of dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 3 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 4 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 5 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 6 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 7 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 8 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention；
Fig. 9 is the structural schematic diagram of another dynamic circulation cell culture apparatus provided in an embodiment of the present invention.
Specific embodiment
With reference to the accompanying drawing to a kind of dynamic circulation cell culture apparatus provided in an embodiment of the present invention and cell culture processes
It is described in detail.
In the description of the present invention, it is to be understood that, term " center ", "upper", "lower", "front", "rear", " left side ",
The orientation or positional relationship of the instructions such as " right side ", "vertical", "horizontal", "top", "bottom", "inner", "outside" is based on the figure
Orientation or positional relationship is merely for convenience of description of the present invention and simplification of the description, rather than the device of indication or suggestion meaning or
Element must have a particular orientation, be constructed and operated in a specific orientation, therefore be not considered as limiting the invention.
Term " first ", " second " be used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance or
Implicitly indicate the quantity of indicated technical characteristic." first " is defined as a result, the feature of " second " can be expressed or imply
Ground includes one or more of the features.
It should be noted that dynamic circulation cell culture apparatus provided in an embodiment of the present invention is suitable for the training of various cells
It supports, according to the different only by environmental parameter needed for changing various cell growths of cell category, for example, each in culture solution
The concentration parameter of nutriment, cytokine concentrations parameter etc.；The embodiment of the present invention is particularly suitable for suspension cell and adherent thin
The culture of born of the same parents.
On the one hand, the embodiment of the present invention provides a kind of dynamic circulation cell culture apparatus, referring to fig. 2, comprising:
Hollow fiber reactor 1, the hollow fiber reactor 1 include culture solution channel 11 and cell growth chamber 12；
Porcine HGF delivery pump 02；The input terminal of the Porcine HGF delivery pump 02 is for inputting cell growth
The factor, the inlet communication of the output end of the Porcine HGF delivery pump 02 and the cell growth chamber 12, the cell are raw
Long factor delivery pump 02 is used to convey Porcine HGF to the cell growth chamber 12.
A kind of dynamic circulation cell culture apparatus provided in an embodiment of the present invention, in cell cultivation process, as cell is raw
Long, growth factor can be constantly consumed, while with the continuous discharge of waste liquid, the Porcine HGF of the cell growth chamber 12
Content can reduce, can be indefinite by the Porcine HGF delivery pump 02 by the way that Porcine HGF delivery pump 02 is arranged
When to the cell growth chamber 12 convey Porcine HGF, keep the Porcine HGF content to be maintained at certain model
In enclosing, so as to improve cell culture effect.
In one embodiment of the invention, referring to Fig. 3, the incubator further includes Porcine HGF storage tank 3, the cell
The outlet of growth factor storage tank 3 is connected to the input terminal of the cell factor delivery pump 02, the Porcine HGF storage tank 3
Bottom is provided with import, and the entrance of the Porcine HGF storage tank 3 is provided with shut-off valve a, and the shut-off valve a is for closing
The Porcine HGF storage tank 3 is isolated from the outside when closing.
In embodiments of the present invention, by the way that shut-off valve a is arranged, it can be avoided extraneous bacterium into the Porcine HGF
In storage tank 3.
Wherein, it should be noted that since hollow fiber reactor generally includes container and setting in the container
At least one hollow fiber conduit, the inside of the hollow fiber conduit and it is external be respectively the culture solution channel 11 and described thin
Intracellular growth chamber 12, for example, when the inside of the hollow fiber conduit is the culture solution channel 11, then the hollow fiber conduit
Outside is the cell growth chamber 12, when the outside of the hollow fiber conduit is culture solution channel 11, then the doughnut
The inside of pipe is the cell growth chamber 12.
It should also be noted that, it is raw suspension cell directly can be inoculated in cell when cultivating suspension cell
In long chamber 12, cell is in suspended state, and when cultivating attached cell, it needs to carry out the surface of cell growth chamber 12
Pretreatment, for example, when the inside of the hollow fiber conduit is cell growth chamber 12, then on the inner wall of the hollow fiber conduit
It is laid with one layer of albumen, the inner wall for enabling cell to be attached to the hollow fiber conduit carries out adhere-wall culture.Vice versa.
In one embodiment of the invention, the incubator further includes pressure extension box 2 and culture solution delivery pump 01, the pressure
Filled with gas, the bottom of the pressure extension box 2 and the culture solution channel 11 or the cell growth chamber in power extension box 2
12 connections；
The input terminal of the culture solution delivery pump 01 for inputting culture solution, output end and the culture solution channel 11 into
Mouth connection, the culture solution delivery pump 01 are used to flowing through culture solution into the culture solution channel 11 in the form of impulse wave, so that
The pressure difference of the culture solution channel 11 and the cell growth chamber 12 is positive and negative alternately to be changed, and the pressure extension box 2 is for leading to
That crosses gas volume changes the pressure balance so that the culture solution channel 11 and the cell growth chamber 12.
By the way that pressure extension box 2 is arranged, and it is filled with a certain amount of gas in the pressure extension box 2, by the pressure
Extension box 2 is connected to the culture solution channel 11 or cell growth chamber 12, and by the culture solution delivery pump 01 with pulse
The form of wave conveys culture solution to the culture solution channel 11, so that the culture solution channel 11 and the cell growth chamber 12
Pressure difference is positive and negative alternately to be changed, when the pressure difference of the culture solution channel 11 and the cell growth chamber 12 is timing, culture solution
The culture solution in channel 11 enters the cell growth chamber 12, and the gas volume in the pressure extension box 2 is compressed into the training
The pressure balance in nutrient solution channel 11 and the cell growth chamber 12, when the culture solution channel 11 and the cell growth chamber 12
When pressure difference is negative, the waste liquid of the cell growth chamber 12 enters the culture solution channel 11, the gas in the pressure extension box 2
Body volume increases the pressure balance until the culture solution channel 11 and the cell growth chamber 12, so as to realize culture solution
With the dynamic exchange of waste liquid；In vivo culture is simulated while being updated to the culture solution of the cell growth chamber 12, is cell
Growth provides stable physiological environment, reduces the consumption of culture solution.
Wherein, it is also necessary to which explanation, for different cell categories, the exchange capacity of culture solution and waste liquid is not yet
Together, and the pressure difference that needs to be applied between the culture solution channel 11 and the cell growth chamber 12 would also vary from,
For example, the attached cell compared with the suspension cell, is attached on the tube wall of the hollow fiber conduit due to attached cell,
Can more or less blocking be generated to the exchange of culture solution and waste liquid, therefore, usual attached cell culture solution channel when cultivating
Pressure difference between 11 and the cell growth chamber 12 is slightly larger than pressure difference when suspension cell culture.
In another embodiment of the present invention, referring to fig. 4, the dynamic circulation cell culture apparatus further includes culture liquid storage tank 4,
The outlet of the culture liquid storage tank 4 is connected to the input terminal of the culture solution delivery pump 01, the outlet in the culture solution channel 11
With the inlet communication of the culture liquid storage tank 4.
In embodiments of the present invention, liquid storage tank 4 is cultivated by setting, culture solution circulation loop is capable of forming, in culture solution
During swapping with waste liquid, the waste liquid of discharge constantly can be taken back institute by the culture solution for flowing through the culture solution channel 11
Culture liquid storage tank 4 is stated, and is used for continuous feed flow, dynamic circulation utilization is carried out to culture solution, reduces culture solution waste, and can protect
The nutriment for being conducive to cell growth in waste liquid is stayed, cell culture is conducive to.
In one embodiment of the invention, referring to Fig. 5, the dynamic circulation cell culture apparatus further includes waste tank 5, waste liquid
Delivery pump 03 and the first sampler 04, the outlet of the input terminal of the waste liquid delivery pump 03 and the culture solution channel 11,
Output end is connected to the waste tank 5, the outlet in the collection terminal of first sampler 04 and the culture solution channel 11 it
Between pipeline connection, the first sampler 04 is for being sampled the culture solution for flowing through the culture solution channel 11.
It is sampled by the culture solution that the first sampler 04 flows out the culture solution channel 11, it can be to circulation loop
In the Metabolites Concentration of culture solution detected in time, so as to according to testing result to the training in the circulation loop
Nutrient solution is handled in time, for example, being unfavorable for cell when the Metabolites Concentration for detecting the culture solution in the circulation loop is higher
When growth, the waste liquid delivery pump 03 can be opened, to collecting the culture solution in the waste tank 5.
In another embodiment of the present invention, referring to Fig. 6, the dynamic circulation cell culture apparatus further includes the second sampler
05, the sampling end of second sampler 05 is connected to the cell growth chamber 12, and second sampler 05 is used for described
Cell and culture solution in cell growth chamber 12 are sampled.
It is sampled by cell and culture solution of second sampler 05 to the cell growth chamber 12, it can be to institute
The cell density for stating cell growth chamber 12 is detected, so as to continue culture or to cell to determine according to testing result
It is acquired, it is, of course, also possible to the Metabolites Concentration in the culture solution of the cell growth chamber 12 is detected, and according to
Testing result performs corresponding processing it, such as: when Metabolites Concentration is excessively high in the culture solution of the cell growth chamber, no
When conducive to cell growth, it can alternately be changed by controlling the pressure difference in the culture solution channel 11 and the cell growth chamber 12
Frequency, to promote exchanging for waste liquid and culture solution, so that the Metabolites Concentration of the cell growth chamber 12 is reduced, it is exemplary
, the frequency of the positive and negative alternately variation of the pressure difference of the culture solution channel 11 and the cell growth chamber 12 can be by controlling institute
The pulse frequency of culture solution delivery pump 01 is stated to realize.
In one embodiment of the invention, referring to Fig. 7, the dynamic circulation cell culture apparatus further includes cellular collector 06,
Between the import of the cellular collector 06 and the Porcine HGF storage tank 3 and the Porcine HGF delivery pump 02
Pipeline connection, the cellular collector 06 are used in 02 antiport of Porcine HGF delivery pump that the cell is raw
Cell collection in long chamber 12 gets up.
In embodiments of the present invention, it is carried out by cell and culture solution of second sampler 05 to the cell growth chamber 12
Sampling, can be by by the Porcine HGF if the cell density reaches certain value after detecting its cell density
02 antiport of delivery pump gets up the cell collection in the cell growth chamber 12.
In one embodiment of the invention, referring to Fig. 8, the Porcine HGF storage tank 3 and the Porcine HGF are defeated
It send and is additionally provided with heater 6 on the pipeline between pump 02, the heater 6 is used for the Porcine HGF for flowing through the pipeline
It is heated.
By the way that heater 6 is arranged, the temperature for the Porcine HGF for entering the cell growth chamber 12 can be adjusted
Section, improves the activity of the Porcine HGF, to improve cell culture effect.
In one embodiment of the invention, the dynamic circulation cell culture apparatus further includes controller (not shown), institute
It is electric with the shut-off valve a, culture solution delivery pump 01, Porcine HGF delivery pump 02, waste liquid delivery pump 03 respectively to state controller
Connection is opened for controlling the shut-off valve a, culture solution delivery pump 01, Porcine HGF delivery pump 02, waste liquid delivery pump 03
It opens and closes, and control the culture solution delivery pump 01 and convey culture solution to the culture solution channel 11 in the form of impulse wave,
Control 02 Positive work of Porcine HGF delivery pump or antiport.By the way that controller is arranged, cell can be trained
Feeding whole process carries out automation control.
In one embodiment of the invention, referring to Fig. 9, the entrance of the culture liquid storage tank 4 is provided with the first valve b, institute
The top for stating culture liquid storage tank 4 is additionally provided with gas vent, the cell growth chamber 12 and the first valve b and the culture solution
Pipeline between channel 11 is connected to by first pipe, and is provided at the cell growth chamber 12 in the first pipe
Exhaust valve c, is additionally provided with the second valve d on the pipeline between the first valve b and the culture solution channel 11, and described first
The outlet of pipeline is connected to the output end side of the second valve d.
Exhaust gas can be generated in cell cultivation process, a part of exhaust gas can enter in the pressure extension box 2, and in addition one
Dividing can smoothly enter into the culture solution when exhaust gas can be opened in the exhaust valve c, the first valve b is opened, the second valve d is closed
In storage tank 4, exhaust gas can be passed through row of exhausting-gas hole to outside by the culture liquid storage tank 4.
On the other hand, the embodiment of the present invention provides a kind of dynamic circulation cell culture processes, is applied to as described above dynamic
State circulating cells incubator, comprising:
Porcine HGF is conveyed to the cell growth chamber by the Porcine HGF delivery pump, so that described thin
The intracavitary Porcine HGF content of intracellular growth is maintained in the first preset range.
A kind of dynamic circulation cell culture processes provided in an embodiment of the present invention are conveyed by setting Porcine HGF
Pump, and with the culture of cell, Porcine HGF sporadically is conveyed into the cell growth chamber, can be prevented because of cell
It grows and constantly consumes Porcine HGF and waste liquid and be constantly discharged so that the Porcine HGF of the cell growth chamber contains
Amount reduces, so that the content of the Porcine HGF keeps in a certain range, can be improved cell culture effect.
In another embodiment of the present invention, the method also includes: by the culture solution delivery pump by culture solution with
The form of impulse wave flows through the culture solution channel, alternately changes so that the pressure difference inside and outside the hollow fiber conduit is positive and negative,
The pressure difference of the culture solution channel and the cell growth chamber is timing, and the culture solution in the culture solution channel enters described thin
Intracellular growth chamber, the gas volume of the pressure extension box are compressed into the pressure in the culture solution channel and the cell growth chamber
Balance；When pressure difference in the culture solution channel and the cell growth chamber is negative, the waste liquid of the cell growth chamber enters
The culture solution channel, the gas volume of the pressure extension box increase to the culture solution channel and the cell growth chamber
Pressure balance.Can promote the exchange of the culture solution and waste liquid, the consumption of less culture solution prevents from cell being grown in waste liquid
Advantageous nutriment it is rapidly losing.
Wherein, it should be noted that for different cell categories, the culture solution channel and the cell growth chamber it
Between pressure difference demand it is different, for example, the attached cell is compared with the suspension cell, since attached cell is attached to
On the tube wall of the hollow fiber conduit, can more or less blocking be generated to the exchange of culture solution and waste liquid, it is therefore, usually adherent thin
Born of the same parents cultivate when the culture solution channel 11 and the cell growth chamber 12 between pressure difference be slightly larger than suspension cell culture when
Pressure difference.
Wherein, pulse, which refers to the process of, rapidly returns back to its original state after a physical quantity is mutated in short duration.
Without limitation to the concrete form of the impulse wave, the impulse wave can be rectangular wave, can be sine wave, or its
The waveform of his shape.
In one embodiment of the invention, it is described culture solution is flowed through in the form of impulse wave culture solution delivery pump it is described
Culture solution channel specifically includes:
The culture solution delivery pump is controlled with the first pre-set velocity and the second pre-set velocity alternately to the culture perfusion
Side conveys culture solution.
In embodiments of the present invention, the culture flow velocity that the culture perfusion side is flowed through by controlling, can be to described
The pressure of culture perfusion side is controlled, and the pressure difference of the culture solution channel and the cell growth chamber is with the shape of rectangular wave
Formula variation, for the speed of the culture solution delivery pump in the wave crest of the rectangular wave, the pressure in the culture solution channel is greater than institute
The pressure of cell growth chamber is stated, culture solution enters the cell growth chamber up to pressure balance, then, the culture solution delivery pump
Speed be transformed into the trough of the rectangular wave rapidly, the pressure of the cell growth chamber is greater than the pressure in the culture solution channel
Power, waste liquid enters the culture solution channel until pressure balance.
Wherein, replace the frequency changed to first pre-set velocity and the second pre-set velocity without limitation, described first
The frequency that pre-set velocity and the second pre-set velocity alternately change can be determined by the cell category being inoculated with, due to different cells
Type, when culture required nutriment concentration therefore difference also has the exchange frequency of culture solution and waste liquid
Institute is different.
In one embodiment of the invention, the method also includes: every the first preset time, pass through first sampler
Detection is sampled to the culture solution for flowing through the culture solution channel, if Metabolites Concentration is more than or equal to the in the culture solution
When one preset value, then the waste liquid delivery pump is controlled by the controller and opened, the culture in the culture solution channel will be flowed through
Liquid drains into the waste tank.
During carrying out Cyclic culture to the cell by culture solution, as the growth of cell can generate largely
Metabolite is sampled detection by the culture solution flowed out to the culture solution channel, and when Metabolites Concentration is excessively high
Control loop circuit is closed, and the culture solution is drained into waste tank, can be timely updated to the environment of cell culture,
Improve the efficiency of cell culture.
In one embodiment of the invention, the method also includes: every the second preset time, pass through second sampler
Cell and culture solution to the cell growth chamber, which are sampled, detects cell density therein, if the cell density
More than or equal to the second preset value, then the Porcine HGF delivery pump antiport is controlled by the controller, it will be described
Cell delivery in cell growth chamber is into the cellular collector.
It is described reversed by the controller control Porcine HGF delivery pump in another embodiment of the present invention
Operating, by the cell delivery in the cell growth chamber into the cellular collector after further include: pass through the controller
The Porcine HGF delivery pump Positive work is controlled, is inoculated with carefully by the cellular collector to the cell growth chamber
Born of the same parents.In embodiments of the present invention, inoculating cell can also be carried out to the cell growth chamber by the cellular collector, promoted
The lasting culture of next wheel cells.
In one embodiment of the invention, the method also includes: every third preset time, controls first valve and open
It opens, the closing of exhaust valve opening and the second valve, portion caused by the cell growth chamber is discharged into the culture solution and is stored up
In tank.
By closing second valve, the portion that the cell growth chamber can be made to generate smoothly drains into described
It cultivates in liquid storage tank, and passes through the row of exhausting-gas hole of the culture solution tank top to outside.
Certainly, the method can also include: every the 4th preset time, by second sampler to the cell
The culture solution of growth chamber is sampled, and detects its Porcine HGF content, if the Porcine HGF content and first
Difference between the lower limit of preset range is less than or equal to the first preset threshold, then controls the Porcine HGF delivery pump and open
It opens, the Porcine HGF of the second predetermined amount is conveyed to the cell growth chamber by Porcine HGF storage tank, so that described
Porcine HGF content is maintained in first preset range.
It is exchanged as cell grows constantly consumption Porcine HGF and culture solution with the continuous of waste liquid, cell growth chamber
Porcine HGF content can reduce, detected by the Porcine HGF content to the cell growth chamber, in institute
Extraneous infection is completely cut off by shut-off valve when stating Porcine HGF content deficiency, it is raw that Xiang Suoshu cell growth chamber supplements steril cell
The long factor promotes cell to grow well.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (10)
1. a kind of dynamic circulation cell culture apparatus characterized by comprising
Hollow fiber reactor, the hollow fiber reactor include culture solution channel and cell growth chamber；
Porcine HGF delivery pump；The input terminal of the Porcine HGF delivery pump is for inputting Porcine HGF, institute
State the output end of Porcine HGF delivery pump and the inlet communication of the cell growth chamber, the Porcine HGF delivery pump
For conveying Porcine HGF to the cell growth chamber；
The incubator further includes pressure extension box and culture solution delivery pump, and the pressure extension box is interior filled with gas, the pressure
The bottom of power extension box is connected to the culture solution channel or the cell growth chamber；
The dynamic circulation cell culture apparatus further includes culture liquid storage tank, and the outlet of the culture liquid storage tank and the culture solution are defeated
The input terminal of pump is sent to be connected to, the inlet communication of the output end of the culture solution delivery pump and the culture solution channel, the culture
The inlet communication of the outlet in liquid channel and the culture liquid storage tank；
Culture solution for being flowed through the culture solution channel by the culture solution delivery pump in the form of impulse wave；The pressure extension
When case changes for the positive and negative alternating of the pressure difference between the culture solution channel and cell growth chamber, pass through the change of gas volume
Change the pressure balance so that the culture solution channel and the cell growth chamber.
2. dynamic circulation cell culture apparatus according to claim 1, which is characterized in that
The incubator further includes Porcine HGF storage tank, the outlet and the cell factor of the Porcine HGF storage tank
The input terminal of delivery pump is connected to, and the bottom of the Porcine HGF storage tank is provided with import, the Porcine HGF storage tank
Entrance be provided with shut-off valve.
3. dynamic circulation cell culture apparatus according to claim 1, which is characterized in that
The dynamic circulation cell culture apparatus further includes waste tank, waste liquid delivery pump and the first sampler, the waste liquid conveying
The outlet of the input terminal of pump and the culture solution channel, output end are connected to the waste tank, first sampler
Collection terminal and the culture solution channel outlet.
4. dynamic circulation cell culture apparatus according to claim 1, which is characterized in that
The dynamic circulation cell culture apparatus further includes the second sampler, and the collection terminal and the cell of second sampler are raw
Long chamber connection.
5. dynamic circulation cell culture apparatus according to claim 4, which is characterized in that
The dynamic circulation cell culture apparatus further includes cellular collector, and the import of the cellular collector and the cell are grown
Pipeline connection between factor storage tank and the Porcine HGF delivery pump, the cellular collector are used for raw in the cell
The cell collection in the cell growth chamber is got up when long factor delivery pump antiport.
6. dynamic circulation cell culture apparatus according to claim 2, which is characterized in that
The dynamic circulation cell culture apparatus further includes controller, and the controller is conveyed with the shut-off valve, culture solution respectively
Pump, the electrical connection of Porcine HGF delivery pump, for controlling the shut-off valve, culture solution delivery pump and Porcine HGF conveying
The opening and closing of pump, and control the culture solution delivery pump culture solution is flowed through into the culture solution in the form of impulse wave and lead to
Road controls the Porcine HGF delivery pump Positive work or antiport.
7. according to the described in any item dynamic circulation cell culture apparatus of claim 3-5, which is characterized in that
The dynamic circulation cell culture apparatus further includes controller, the controller respectively with the culture solution delivery pump, cell
Growth factor delivery pump and the electrical connection of waste liquid delivery pump, for controlling culture solution delivery pump, Porcine HGF delivery pump and giving up
The opening and closing of liquid delivery pump, and control the culture solution delivery pump and culture solution is flowed through to the culture in the form of impulse wave
Liquid channel controls the Porcine HGF delivery pump Positive work or antiport.
8. a kind of dynamic circulation cell culture processes are applied to the described in any item dynamic circulation cell culture of claim 1-7
Device characterized by comprising
Porcine HGF is conveyed to the cell growth chamber by the Porcine HGF delivery pump, so that the cell is raw
Long intracavitary Porcine HGF content is maintained in the first preset range；
And culture solution is flowed through culture solution delivery pump by the culture solution channel in the form of impulse wave, so that the culture
The positive and negative alternately variation of pressure difference between liquid channel and cell growth chamber；
Change the pressure so that the culture solution channel and the cell growth chamber by the gas volume in pressure extension box
Balance, promotes the culture solution for flowing through the culture solution channel and the waste liquid generated in cell growth chamber to swap.
9. dynamic circulation cell culture processes according to claim 8, which is characterized in that the dynamic circulation cell culture
Device includes waste tank, waste liquid delivery pump, the first sampler and controller, and the waste liquid delivery pump and the controller are electrically connected
It connects, the outlet of the collection terminal of first sampler and the culture solution channel；
The method also includes: every the first preset time, by first sampler to flowing through the culture solution channel
Culture solution is sampled detection, if Metabolites Concentration is more than or equal to the first preset value in the culture solution, by described
Controller controls waste liquid delivery pump and opens, and the culture solution for flowing through the culture solution channel is drained into the waste tank.
10. dynamic circulation cell culture processes according to claim 8, which is characterized in that the dynamic circulation cell training
Feeding device includes controller and the second sampler, and the controller is electrically connected with the Porcine HGF delivery pump, and described
The collection terminal of two samplers is connected to the cell growth chamber；
The method also includes: every the second preset time, by second sampler to the cell of the cell growth chamber
It is sampled with culture solution and cell density therein is detected, if the cell density is more than or equal to the second preset value,
The Porcine HGF delivery pump antiport is controlled by the controller, by the cell delivery in the cell growth chamber
Into cellular collector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610800907.5A CN106497783B (en) | 2016-09-01 | 2016-09-01 | A kind of dynamic circulation cell culture apparatus and cell culture processes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610800907.5A CN106497783B (en) | 2016-09-01 | 2016-09-01 | A kind of dynamic circulation cell culture apparatus and cell culture processes |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106497783A CN106497783A (en) | 2017-03-15 |
| CN106497783B true CN106497783B (en) | 2019-03-22 |
Family
ID=58291330
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610800907.5A Active CN106497783B (en) | 2016-09-01 | 2016-09-01 | A kind of dynamic circulation cell culture apparatus and cell culture processes |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106497783B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107189975B (en) * | 2017-05-26 | 2020-12-25 | 奥凯（苏州）生物技术有限公司 | Cell collection method of closed cell amplification system |
| CN107058086B (en) * | 2017-05-26 | 2020-07-03 | 奥凯（苏州）生物技术有限公司 | Method and system for preventing cells from blocking culture device |
| CN107043701A (en) * | 2017-05-26 | 2017-08-15 | 奥凯（苏州）生物技术有限公司 | A kind of cell expansion system for improving cell inoculation dispersiveness |
| CN111592985B (en) * | 2020-04-28 | 2024-02-06 | 郑州伊美诺生物技术有限公司 | Continuous culture device and method for hybridoma cells |
| CN113817605A (en) * | 2021-09-18 | 2021-12-21 | 河南科技大学 | Dynamic cell culture device and single cell culture device |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2206892A (en) * | 1987-07-07 | 1989-01-18 | Aftab Alam | Stationary cell culture system |
| FR2794130B1 (en) * | 1999-05-26 | 2003-01-03 | Bertin Technologies Sa | METHOD AND DEVICE FOR CULTURING MULTI-APPLICATION CELLS |
| US20030054544A1 (en) * | 2001-09-14 | 2003-03-20 | Medcell Biologics, Inc. | Oxygen enriched bioreactor and method of culturing cells |
| CN100376665C (en) * | 2005-12-15 | 2008-03-26 | 中国人民解放军军事医学科学院野战输血研究所 | Cell culturation apparatus |
| CN201737949U (en) * | 2010-07-12 | 2011-02-09 | 中国人民解放军第三军医大学第一附属医院 | Cell culturing device capable of periodically converting positive pressure and negative pressure |
| ES2508615T3 (en) * | 2010-08-06 | 2014-10-16 | Tissuse Gmbh | Circulation system |
| GB201019932D0 (en) * | 2010-11-24 | 2011-01-05 | Imp Innovations Ltd | Dimensional hollow fibre bioreactor systems for the maintenance, expansion, differentiation and harvesting of human stem cells and their progeny |
| EP2639294A1 (en) * | 2012-03-15 | 2013-09-18 | CellProthera | Automaton and automated method for cell culture |
| CN203048948U (en) * | 2012-10-17 | 2013-07-10 | 浙江赛尚医药科技有限公司 | Cell culture piping device and cell automated culture system |
| CN103589808A (en) * | 2013-11-11 | 2014-02-19 | 中国人民解放军第三军医大学第一附属医院 | Automatic control method and system for pH/oxygen partial pressure/partial pressure of carbon dioxide of cell and tissue culture solution |
| CN103614296B (en) * | 2013-11-15 | 2015-05-20 | 汪洋 | Double-chamber three-dimensional pouring bioreactor system |
-
2016
- 2016-09-01 CN CN201610800907.5A patent/CN106497783B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN106497783A (en) | 2017-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106497783B (en) | A kind of dynamic circulation cell culture apparatus and cell culture processes | |
| CN106635790B (en) | A kind of fully enclosed automatic cell culture system | |
| CN106467892B (en) | A kind of automatic cytological amplification system | |
| CN106479889B (en) | A kind of cell culture circulatory system | |
| CN102382766B (en) | Tidal bioreactor for animal cell culture | |
| CN106497784B (en) | A kind of control pressurer system and its compress control method for realizing culture solution and waste liquid exchange | |
| CN206783688U (en) | A kind of multilevel cell amplification system | |
| CN105543085B (en) | The micro fluidic device and method controlled for microorganism fertilizer bacterium training systern | |
| CN107904169A (en) | A kind of modularized bioreactor suitable for various kinds of cell type culture | |
| CN106520552B (en) | A kind of biological reactor for cell culture | |
| JP2018529367A (en) | Bioreactor capable of shutting off gas supply | |
| CN206014949U (en) | A kind of nutrition liquid supplying device for cell culture | |
| CN109825436A (en) | Totally-enclosed cell culture system | |
| CN107236669A (en) | A kind of the high efficiency culture systems and application process of mass cell production and culture | |
| CN102199537A (en) | Membrane bioreactor used in microgravity environment and simulated microgravity environment | |
| CN111944692B (en) | Animal cell hollow fiber culture system and cell culture method thereof | |
| CN203065461U (en) | Internal loop airlift microbial culture reactor | |
| CN206843495U (en) | Ezyme bio-reactor and intelligent enzyme reaction work station | |
| CN102002481A (en) | Production method of porcine reproductive and respiratory syndrome virus | |
| CN102002482B (en) | Method for producing PRRS (Porcine Reproductive and Respiratory Syndrome) viruses | |
| CN104974933B (en) | A kind of extensive continuous several times, which suspend, turns the apparatus and method of expression recombinant protein wink | |
| CN206814791U (en) | Disposable intelligent digester systems for animal cell culture | |
| CN102154100B (en) | Potential difference circulating bioreactor | |
| CN106834117B (en) | For releasing the enzyme reactor and intelligent enzyme reaction work station of zooblast attaching | |
| CN208667686U (en) | A kind of fermented tea installation for fermenting |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |