CN106467892B - A kind of automatic cytological amplification system - Google Patents
A kind of automatic cytological amplification system Download PDFInfo
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- CN106467892B CN106467892B CN201610800908.XA CN201610800908A CN106467892B CN 106467892 B CN106467892 B CN 106467892B CN 201610800908 A CN201610800908 A CN 201610800908A CN 106467892 B CN106467892 B CN 106467892B
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Abstract
The present invention relates to technical field of cell culture more particularly to a kind of automatic cytological amplification systems.It can be realized and automation real-time monitoring is carried out to the gas meltage in cell growing environment, provide stable growing environment for cell culture.A kind of automatic cytological amplification system, comprising: the culture solution conveying device and hollow fiber reactor, the hollow fiber reactor being connected to by the circulation conduit of culture solution include culture solution channel and cell growth chamber；The culture solution conveying device includes gas dissolver, the gas dissolver includes gas feed, culture solution import and the culture solution outlet of shell and setting on the housing, and the gas dissolver forms circulation loop with the culture solution channel by the culture solution import and culture solution outlet；The gas being connected to the gas feed provides device, and the gas provides and is provided with gas flow regulating valve between device and the gas feed；The control system being electrically connected with the gas flow regulating valve.
Description
Technical field
The present invention relates to technical field of cell culture more particularly to a kind of automatic cytological amplification systems.
Background technique
T cell has multiple biological function, and such as direct killing target cell, auxiliary or inhibition B cell generate antibody, to spy
The responsing reaction of Specific Antigen and mitogenesis original and generate cell factor etc., mature T cell is through blood distribution to outer
The thymus dependent area of all immune organs is settled down, and can be recycled through lymphatic vessel, peripheral blood and tissue fluid etc., is played cell and is exempted from
The functions such as epidemic disease and immunological regulation.Successful incubation T cell, and this cell is largely injected into patient's body, to enhance immune system
As a kind of possibility.
T cell cultivate mainly using two methods in the prior art, one kind is to use Tissue Culture Dish, high-grade
Laminar flow hood and carbon dioxide incubator cultivated, this method efficiency is lower, is not able to satisfy clinical demand；Another kind is logical
It crosses hollow fiber reactor to cultivate the cells, one of culture apparatus is sequentially connected referring specifically to Fig. 1 by diversion pipe
Main circulation pump 1, gas exchanges pipe 2, culture tube 3 and culture solution master library 4, be provided on the culture solution master library 4 with it is extraneous even
Logical gas vent；Wherein, the culture tube 3 is hollow fiber reactor, and the diversion pipe includes connecting 1 He of main circulation pump
First diversion pipe 11 of the gas exchanges pipe 2, the connection gas exchanges pipe 2 and the culture tube 3 the second diversion pipe 12,
It connects the third diversion pipe 13 of the culture tube 3 and the culture solution master library 4 and connects the culture solution master library 4 and the master
4th diversion pipe 14 of circulating pump 1, the third diversion pipe 13 are connected to by the 5th diversion pipe 15 with waste tank 5, wherein institute
The cylindrical shell that hollow fiber reactor 3 includes hollow fiber conduit and the installation hollow fiber conduit is stated, it is described hollow
The inside and outside of fibre pipe is respectively formed culture solution channel 31 and cell growth chamber 32, flows through institute in the culture solution channel 31
Culture solution, inoculating cell in the cell growth chamber 32 are stated, the cell growth chamber 32 passes through the 6th with the 5th diversion pipe
Diversion pipe 16 is connected to, and it is the cell that culture solution can enter in the cell growth chamber 32 through the aperture of the hollow fiber conduit
Nutrition is provided to cultivate the cells, waste liquid caused by cell culture can drain into waste tank 5 through the 6th diversion pipe 16,
During this, cell culture system is closed system, is also difficult in the prior art in the closed system in cell growth process
The meltage (such as: oxygen, carbon dioxide etc.) of gas is suitably adjusted, so that cell is difficult under conditions of being suitable for stablizing
Growth, therefore, the regulation of cell growing environment constrains application of the hollow fiber reactor in terms of cell culture.
Summary of the invention
The embodiment of the present invention provides a kind of automatic cytological amplification system, can be realized to the gas in cell growing environment
Body meltage carries out automation real-time monitoring, provides stable growing environment for cell culture.
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 automatic cytological amplification system, comprising:
The culture solution conveying device and hollow fiber reactor being connected to by the circulation conduit of culture solution, the hollow fibre
Tieing up reactor includes culture solution channel and cell growth chamber；The culture solution conveying device includes gas dissolver, the gas
Dissolvers includes gas feed, culture solution import and the culture solution outlet of shell and setting on the housing, the gas
Dissolvers forms circulation loop with the culture solution channel by the culture solution import and culture solution outlet；
The gas being connected to the gas feed provides device, and the gas provides to be set between device and the gas feed
It is equipped with gas flow regulating valve；
The control system being electrically connected with the gas flow regulating valve；
The culture solution conveying device is used to convey culture solution to the culture solution channel, and the control system is for controlling
The aperture of the gas flow regulating valve to the gas provide device be passed through into the gas dissolver flow of gas into
Row is adjusted.
Preferably, it is provided with hollow fiber conduit in the shell of the gas dissolver, the hollow fiber conduit is by the shell
Internal space is divided into culture solution and flows through side and gas flow side, and the culture solution flows through side and the culture solution import and described
Culture solution outlet, the gas flow side are connected to the gas feed.
Optionally, it includes dioxide bottle and oxygen gas cylinder that the gas, which provides device, the dioxide bottle with
It is provided with first flow regulating valve between the gas feed, is provided with second between the oxygen gas cylinder and the gas feed
Flow control valve, the control system are electrically connected with the first flow regulating valve and the second flow regulating valve, for pair
The flow for the carbon dioxide and oxygen being passed through in the gas dissolver is adjusted.
Further, the control system is for obtaining the consumption models of carbon dioxide and the consumption models of oxygen, and root
The aperture of the first flow regulating valve is adjusted according to the consumption models of the carbon dioxide, so that the pH value of culture solution is protected
It holds in the first preset range, is adjusted according to aperture of the consumption models of the oxygen to the second flow regulating valve,
So that the dissolved oxygen amount of culture solution is maintained in the second preset range.
Optionally, be provided with pH detector and dissolved oxygen amount detector on the circulation conduit, the control system with it is described
PH detector and dissolved oxygen amount detector electrical connection, the control system are used for the pH signal sent according to the pH detector
The aperture of the first flow regulating valve is adjusted, so that pH value is maintained in the first preset range；According to the dissolved oxygen
The aperture of the second flow regulating valve is adjusted in the dissolved oxygen amount signal that amount detector is sent, so that dissolved oxygen amount is maintained at the
In two preset ranges.
Preferably, the culture solution conveying device further includes circulating pump, the output end of the circulating pump and the culture solution
Inlet communication, the outlet of input terminal and the culture solution channel, the circulating pump are electrically connected with the control system；
The system also includes pressure extension box；Filled with gas, the bottom of the pressure extension box in the pressure extension box
Portion is connected to the culture solution channel or the cell growth chamber, and the control system is also used to control the circulating pump and will train
Nutrient solution flows through the culture solution channel in the form of impulse wave, so that the pressure in the culture solution channel and the cell growth chamber
Difference is positive and negative alternately to be changed, and the pressure extension box is used to change by gas volume so that the culture solution channel and the cell
The pressure balance of growth chamber.
Further, the cell growth chamber is provided with first pressure sensor, and the culture solution of the gas dissolver goes out
Be additionally provided with second pressure sensor on circulation conduit mouthful between the import in the culture solution channel, the control system with
The first pressure sensor and second pressure sensor electrical connection, it is raw that the control system is also used to obtain the cell
The Metabolites Concentration model of growth of long chamber, and sent according to the first pressure sensor and the second pressure sensor
Pressure controls the circulating pump and conveys culture solution to the culture solution channel with predetermined pulse model, so that the cell growth chamber
Metabolites Concentration be maintained in third preset range.
Optionally, the system also includes waste liquid delivery pump and waste tanks；
The culture solution conveying device further includes the training being arranged between the hollow fiber reactor and the circulating pump
Nutrient solution surge tank, the outlet of the import of the culture solution surge tank and the culture solution channel, outlet and the circulating pump
Input terminal connection；
Pipeline between the input terminal of the waste liquid delivery pump and the culture solution surge tank and the culture solution channel connects
Logical, output end is connected to the waste tank, and the control system is electrically connected with the waste liquid delivery pump；
The control system is also used to obtain the model of growth of the products of cellular metabolism in the culture solution channel, however, it is determined that thin
Difference between the concentration of the products of cellular metabolism in the preset threshold of born of the same parents' metabolite and the culture solution channel is less than or equal to the
When one preset threshold, then controls the waste liquid delivery pump and open, the culture solution in the culture solution channel is delivered to waste tank
In.
Preferably, the system also includes sealant supplement cylinders and culture solution delivery pump；
The cell growth chamber further includes import, and the outlet of the sealant supplement cylinder connects with the input terminal of the culture solution delivery pump
It is logical, the inlet communication of the output end of the culture solution delivery pump and the cell growth chamber, the culture solution of the gas dissolver
It exports and is additionally provided with temperature sensor, the sealant supplement cylinder and the culture solution on the circulation conduit being connected to the culture solution channel
Be additionally provided with heater on the pipeline of delivery pump connection, the control system and the culture solution delivery pump, temperature sensor and
Heater electrical connection；
The control system is also used to obtain the temperature signal that the temperature sensor is sent, and conveys to the culture solution
The open and close of pump and the heater are controlled, so that the temperature in the culture solution channel is maintained at the 4th preset range
It is interior.
Preferably, third is additionally provided on the pipeline of the culture solution delivery pump and the inlet communication of the cell growth chamber
Pressure sensor, the control system are electrically connected with the third pressure sensor, and the control system is also used to according to
The pressure signal that third pressure sensor is sent controls the speed of culture solution delivery pump conveying culture solution, so that institute
The speed for stating culture solution delivery pump conveying culture solution is maintained in the 5th preset range.
The embodiment of the present invention provides a kind of automatic cytological amplification system, provides device by setting gas, and will be described
Gas provides device and is connected to the air inlet, can provide required gas for the gas flow side of the gas dissolver
Body, also, the aperture of the gas flow regulating valve is controlled by control system, it can be to the gas for flowing through the gas flow side
The flow of body is controlled, and stable gas is provided for cell culture, so as to the gas meltage in the culture solution
Automation control is carried out, so that culture solution suitable cell culture, stable growing environment is provided for cell culture.
Detailed description of the invention
Fig. 1 is a kind of structural schematic diagram for cell cycle culture systems that the prior art provides；
Fig. 2 is a kind of structural schematic diagram of automatic cytological amplification system provided in an embodiment of the present invention；
Fig. 3 is the structural schematic diagram of another automatic cytological amplification system provided in an embodiment of the present invention；
Fig. 4 is the structural schematic diagram of another automatic cytological amplification system provided in an embodiment of the present invention；
Fig. 5 is the structural schematic diagram of another automatic cytological amplification system provided in an embodiment of the present invention；
Fig. 6 is the structural schematic diagram of another automatic cytological amplification system provided in an embodiment of the present invention；
Fig. 7 is the structural schematic diagram of another automatic cytological amplification system provided in an embodiment of the present invention.
Specific embodiment
With reference to the accompanying drawing to a kind of automatic cytological amplification system provided in an embodiment of the present invention and its cell culture side
Method 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 a kind of automatic cytological amplification system provided in an embodiment of the present invention is suitable for any cell
Amplification, is particularly suitable for the amplification cultivation of suspension cell and attached cell.
On the one hand, the embodiment of the present invention provides a kind of automatic cytological amplification system, referring to fig. 2, comprising:
The culture solution conveying device and hollow fiber reactor 3 being connected to by the circulation conduit of culture solution, it is described hollow
Fiber reactor 3 includes culture solution channel 31 and cell growth chamber 32；The culture solution conveying device includes gas dissolver 2,
The gas dissolver 2 includes gas feed, culture solution import and the culture solution outlet of shell and setting on the housing,
The gas dissolver 2 forms circulation loop with the culture solution channel 31 by the culture solution import and culture solution outlet；
The gas being connected to the gas feed provides device (in figure shown in 01 and 02), and the gas provides device and institute
It states and is provided with gas flow regulating valve between gas feed (in figure shown in a and b)；
The control system 100 being electrically connected with the gas flow regulating valve；
The culture solution conveying device is used to convey culture solution to the culture solution channel 31, and the control system 100 is used
Device, which is provided, to the gas in the aperture for controlling the gas flow regulating valve is passed through gas into the gas dissolver 2
Flow is adjusted.
The embodiment of the present invention provides a kind of automatic cytological amplification system, provides device by setting gas, and will be described
Gas provides device and is connected to the air inlet, can provide required gas for the gas flow side of the gas dissolver 2
Body, also, control the aperture of the gas flow regulating valve (a and b) by control system 100, can be to flowing through the gas
The flow of the gas of dissolvers 2 is controlled, and certain gas flow is provided for cell culture, so as to the culture solution
In gas meltage carry out automation control so that culture solution suitable cell culture, stable growth is provided for cell culture
Environment.
In one embodiment of the invention, referring to Fig. 3, it is provided with hollow fiber conduit 21 in the shell of the gas dissolver 2,
Space in the shell is divided into culture solution and flows through side 22 and gas flow side 23, the culture solution by the hollow fiber conduit 21
Side 22 and the culture solution import and the culture solution outlet are flowed through, the gas flow side 23 connects with the gas feed
It is logical.
By the way that hollow fiber conduit 21 is arranged, so that the gas in gas flow side 23 is spread by the hollow fiber conduit 21
Enter the culture solution and flow through side 22, can be come into full contact with culture solution, improves the meltage of gas.
In another embodiment of the present invention, it includes dioxide bottle 01 and oxygen gas cylinder 02 that the gas, which provides device,
First flow regulating valve a, the oxygen gas cylinder 02 and institute are provided between the dioxide bottle 01 and the gas feed
It states and is provided with second flow regulating valve b between gas feed, the control system 100 and the first flow regulating valve a and institute
Second flow regulating valve b electrical connection is stated, is carried out for the flow to the carbon dioxide and oxygen being passed through in the gas dissolver 2
It adjusts.
By setting dioxide bottle and oxygen gas cylinder, carbon dioxide gas and oxygen can be provided for the culture solution
Gas meets cell culture demand so as to provide carbon dioxide and oxygen for the culture solution, meanwhile, pass through the control
The gas flow being passed through in the gas dissolver 2 is adjusted in system, can meltage to carbon dioxide and oxygen into
Row control.
In one embodiment of the invention, the control system 100 is used to obtain the consumption models and oxygen of carbon dioxide
Consumption models, and be adjusted according to aperture of the consumption models of the carbon dioxide to the first flow regulating valve a, so that
The pH value of culture solution is maintained in the first preset range, according to the consumption models of the oxygen to the second flow regulating valve b
Aperture be adjusted so that the dissolved oxygen amount of culture solution is maintained in the second preset range.
It, can be according to any time since in cell cultivation process, carbon dioxide primarily serves the effect for adjusting pH value
Solubility of the situation of change and carbon dioxide of the pH value of culture solution in the culture solution controls the logical of the carbon dioxide
Enter amount, carries out real-time monitoring so as to the pH value to culture solution；Meanwhile cell can consume a small amount of oxygen during training oxygen
Gas can control the logical of oxygen according to the solubility of the keto consumpting speed of the cell of any time and oxygen in the culture solution
Enter amount, so as to timely dissolved oxygen, controls dissolved oxygen amount.
In one embodiment of the invention, referring to Fig. 3, pH detector 03 and dissolved oxygen amount detection are provided on the circulation conduit
Device 04, the control system 100 are electrically connected with the pH detector 03 and dissolved oxygen amount detector 04, and the control system 100 is used
It is adjusted in aperture of the pH signal sent according to the pH detector 03 to the first flow regulating valve a, so that pH value
It is maintained in the first preset range；The second flow is adjusted according to the dissolved oxygen amount signal that the dissolved oxygen amount detector 04 is sent
The aperture of valve b is adjusted, so that dissolved oxygen amount is maintained in the second preset range.
By the pH detector 03 and the dissolved oxygen amount detector 04 to the pH value of the culture solution in circulating system and molten
Oxygen amount is measured in real time, and is adjusted in time by the control system 100 to the flow of carbon dioxide and the flow of oxygen
Section, so as to the culture solution in circulating system pH value and dissolved oxygen amount monitor in real time, it is suitable to guarantee to provide for cell
Growing environment.
In one embodiment of the invention, referring to fig. 4, the culture solution conveying device further includes circulating pump 1, the circulating pump
1 output end and the culture solution inlet communication, the outlet of input terminal and the culture solution channel 31, the circulating pump 1
It is electrically connected with the control system 100；
The cell culture circulatory system further includes pressure extension box 6；It is described filled with gas in the pressure extension box 6
The bottom of pressure extension box 6 is connected to the culture solution channel 31 or the cell growth chamber 32, and the control system 100 is also
Culture solution is flowed through to the culture solution channel 31 in the form of impulse wave for controlling the circulating pump 1, so that the culture solution
The pressure difference of channel 31 and the cell growth chamber 32 is positive and negative alternately to be changed, and the pressure extension box 6 is for passing through gas volume
Change the pressure balance so that the culture solution channel 31 and the cell growth chamber 32.
In embodiments of the present invention, by be arranged pressure extension box 6, and by the control system control loop pump 1 with
The form of impulse wave conveys culture solution, so that the positive and negative alternating of pressure difference in the culture solution channel 31 and the cell growth chamber 32
Variation.When the pressure difference is timing, the culture solution in the culture solution channel 31 enters the cell growth chamber 32, pressure extension
The gas volume of case 6 is compressed, until pressure balance.Waste liquid when the pressure difference is negative, in the cell growth chamber 32
Into the culture solution channel 31, the gas volume of pressure extension box 6 increases, until pressure balance, thus realize culture solution and
The exchange of waste liquid, in this way, growing advantageous nutriment to cell in waste liquid and can be cultured liquid band to enter circulating system is cell
It is used, additionally it is possible to constantly to carry out culture solution update, reduce the consumption of culture solution.
For different cell categories, the exchange capacity of culture solution and waste liquid is also different, and is applied to the training
Pressure difference between nutrient solution channel 31 and the cell growth chamber 32 would also vary from, with the attached cell and the suspension
It is illustrated for cell, when cultivating suspension cell, suspension cell directly can be inoculated in cell growth chamber 32
Interior, cell is in suspended state, and when cultivating attached cell, it needs to locate the surface of cell growth chamber 32 in advance
Reason, for example, being then laid on the inner wall of the hollow fiber conduit when the inside of the hollow fiber conduit is cell growth chamber 32
One layer of albumen, the inner wall for enabling cell to be attached to the hollow fiber conduit carry out adhere-wall culture.Vice versa.Therefore, lead to
The exchange of culture solution and waste liquid, the culture solution channel 31 and the cell growth chamber 32 are realized when normal attached cell culture
Between pressure difference be slightly larger than suspension cell culture when pressure difference.
In another embodiment of the present invention, referring to fig. 4, the cell growth chamber 32 is provided with first pressure sensor 05,
The culture solution of the gas dissolver 2, which is exported, is additionally provided with the on the circulation conduit between the import in the culture solution channel 31
Two pressure sensors 06, the control system 100 and 06 electricity of the first pressure sensor 05 and the second pressure sensor
Connection, the control system 100 are also used to obtain the Metabolites Concentration model of growth of the cell growth chamber 32, and according to institute
The pressure for stating first pressure sensor 05 and the second pressure sensor 06 transmission controls the circulating pump 1 with predetermined pulse mould
Type conveys culture solution to the culture solution channel 31, so as to be maintained at third pre- for the Metabolites Concentration of the cell growth chamber 32
If in range.
While by realizing exchange to culture solution and waste liquid, the circulating pump 1 is controlled by culture solution with predetermined pulse mould
Type flows through the culture solution channel 31, can exchange capacity to culture solution and waste liquid and exchange frequency control, to make
The Metabolites Concentration in the cell growth chamber is obtained to be unlikely to excessively high and have a negative impact to cell growth.Illustratively,
The impulse wave of the predetermined pulse model can for rectangular wave, sine wave etc., the wave crest and trough of the predetermined pulse model it
Between difference it is also changeable.The wave crest of exchange capacity in the unit time of culture solution and waste liquid and the predetermined pulse model and
The size and pulse frequency of trough have relationship, can rule of thumb or theoretical calculation determines the predetermined pulse model
Concrete form reduce the metabolism in the cell growth chamber 32 as long as culture solution and waste liquid is enabled to be exchanged in time
Production concentration.
It should also be noted that, metabolite is increasing, so that in the cell growth chamber with the culture of cell
Culture solution viscosity increases, can also be by the way that the impulse wave is gradually shortened in order to improve the exchange effect of culture solution and waste liquid
Time cycle or/and increase the pressure difference between the culture solution channel and the cell growth chamber to realize.
In another embodiment of the present invention, it is anti-that the control system 100 is also used to receive the second pressure sensor 06
The pressure signal of feedback, and be corrected according to pulse signal of the pressure signal of feedback to the circulating pump 1.
In embodiments of the present invention, by closed-loop control, so that the circulating pump 1 is transported with stable pulse signal
Row, and to the pressure difference steady change in the culture solution channel 31 and the cell growth chamber 32.
In one embodiment of the invention, referring to Fig. 5, the top of the pressure extension box 6 is provided with first pipe 61, described
Third valve c is provided in first pipe 61；
6 outside of pressure extension box is provided with the first liquid level biography at the first height and the second height respectively from top to bottom
Sensor 07 and the second liquid level sensor 08, the control system 100 are passed with first liquid level sensor 07, the second liquid level respectively
Sensor 08 and third valve c electrical connection, the control system 100 are used to detect culture solution in first liquid level sensor 07
Liquid level control the third valve c when reaching the first height and open, the gas into the pressure extension box 6, described
Second liquid level sensor 08 detects that controlling the third valve c when liquid level of culture solution reaches the second height opens, from
Discharge section gas in the pressure extension box 6, so that the liquid level of the culture solution is maintained at the first height and second high
Between degree.
It in embodiments of the present invention, can be to institute by the first liquid level sensor 07 and second liquid level sensor 08
The gas flow stated in pressure extension box 6 is monitored in time, so that the gas volume in the pressure extension box 6 is maintained at certain
In the range of, it is controlled convenient for the exchange capacity to culture solution and waste liquid.
In one embodiment of the invention, the system also includes waste tank 5 and waste liquid delivery pump 51,
The culture solution conveying device further includes being arranged between the hollow fiber reactor 3 and the circulating pump 1
Culture solution surge tank 4, the outlet of the import of the culture solution surge tank 4 and the culture solution channel 31, outlet with it is described
The input terminal of circulating pump 1 is connected to；
Pipe between the input terminal of the waste liquid delivery pump 51 and the culture solution surge tank 4 and the culture solution channel 31
Road connection, output end are connected to the waste tank 5, and the control system 100 is electrically connected with the waste liquid delivery pump 51；
The control system 100 is also used to obtain the model of growth of the products of cellular metabolism in the culture solution channel 31, if
Determine that the difference between the preset threshold of products of cellular metabolism and the concentration of the products of cellular metabolism in the culture solution channel 31 is small
When being equal to the first preset threshold, then controls the waste liquid delivery pump 51 and open, the culture solution in the culture solution channel 31 is defeated
It send into waste tank 5.
Wherein, the model of growth of the products of cellular metabolism in the culture solution channel 31 can be obtained by experience, can also be with
It is obtained by experiment, when being obtained by experiment, change curve of the products of cellular metabolism at any time in section can be drawn, led to
The change curve is crossed, any time can be calculated according to the growth rate and any time period of the Metabolites Concentration of any time
Metabolite concentration, however, it is determined that it is default that difference between the concentration and preset threshold of the metabolite is less than or equal to first
When threshold value, then controls the waste liquid delivery pump 51 and open, culture solution is delivered in the waste tank 5.As cell is grown
Progress, the continuous exchange of culture solution and waste liquid, when the Metabolites Concentration in the culture solution in the culture solution channel 31 is higher,
When being unsuitable for cell growth, it can be drained into the waste tank 5.
In another embodiment of the present invention, referring to Fig. 6, the system also includes sealant supplement cylinders 7 and culture solution delivery pump 71；
The cell growth chamber 32 further includes import, and exporting for the sealant supplement cylinder 7 is defeated with the culture solution delivery pump 71
Enter end connection, the inlet communication of the output end of the culture solution delivery pump 71 and the cell growth chamber 32, the gas dissolution
Temperature sensor 09, the benefit are additionally provided on the circulation conduit that the culture solution outlet of device 2 is connected to the culture solution channel 31
Heater 8, the control system 100 and the training are additionally provided on the pipeline that flow container 7 is connected to the culture solution delivery pump 71
Nutrient solution delivery pump 71, temperature sensor 09 and heater 8 are electrically connected；
The control system 100 is also used to obtain the temperature signal that the temperature sensor 09 is sent, and to the culture
The open and close of liquid delivery pump 71 and the heater 8 are controlled, so that the temperature in the culture solution channel 31 is maintained at
In 4th preset range.
During Cyclic culture, in the temperature and the hollow fiber reactor 3 that the culture solution channel 31 is embodied
The temperature in portion is almost the same, is detected by the temperature to circulating system, and passes through control institute according to detected temperature
It states culture solution delivery pump 71 to convey culture solution and heat it or not heater, the culture solution delivery pump 71 can be conveyed
The temperature of culture solution be adjusted, and pass through the friendship in the culture solution channel 31 and the culture solution of the cell growth chamber 32
It changes, the temperature of the circulating system can be adjusted, so that cell is grown in the environment of proper temperature.
In another embodiment of the present invention, the inlet communication of the culture solution delivery pump 71 and the cell growth chamber 32
Third pressure sensor 010 is additionally provided on pipeline, the control system 100 is electrically connected with the third pressure sensor 010,
The control system 100 is also used to convey the culture solution according to the pressure signal that the third pressure sensor 010 is sent
The speed of 71 conveying culture solutions of pump is controlled, so that the speed of the 1 culture solution delivery pump 71 conveying culture solution is maintained at the
In five preset ranges.
In embodiments of the present invention, culture solution is conveyed to the culture solution delivery pump 72 by third pressure sensor 010
Pressure detected in time, can be by the control system 100 to the culture solution channel 31 and the cell growth chamber
32 pressure difference is adjusted, and controls so as to the exchange frequency and exchange capacity to culture solution and waste liquid, realizes to institute
State the accurate regulation of the temperature of circulating system.
In one embodiment of the invention, referring to Fig. 7, the system also includes fluid infusion pump 72, the input of the fluid infusion pump 72
End is connected to the sealant supplement cylinder 7, the inlet communication of output end and the culture solution surge tank 4, and the culture solution surge tank 4 is from upper
It is provided with third liquid level sensor 011 and the 4th liquid level sensor 012 at third height and the 4th height respectively to lower, it is described
Control system 100 is also used to the third liquid level sensor 011 and detects that the liquid level of culture solution reaches third height time control
It makes the fluid infusion pump 72 to close, stops the fluid infusion into the culture solution surge tank 4；It is detected in the 4th liquid level sensor 010
Fluid infusion pump 72 is controlled when reaching four height to the liquid level of culture solution to open, and is mended into the culture solution surge tank 4
Liquid, so that the liquid level of culture solution is maintained between third height and the 4th height in the culture solution surge tank 4.
When carrying out fluid infusion to the culture solution channel 32 and/or the cell growth chamber 33 by the sealant supplement cylinder 7, lead to
It crosses and the liquid level of the culture solution surge tank 4 is monitored, it is insufficient to can be avoided culture solution in the culture solution surge tank 4
Cycle interruption is caused, can also prevent culture solution in the culture solution surge tank 4 from excessively overflowing.
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 automatic cytological amplification system characterized by comprising
The culture solution conveying device and hollow fiber reactor, the doughnut being connected to by the circulation conduit of culture solution are anti-
Answering device includes culture solution channel and cell growth chamber；The culture solution conveying device includes gas dissolver, the gas dissolution
Device includes gas feed, culture solution import and the culture solution outlet of shell and setting on the housing, the gas dissolution
Device forms circulation loop with the culture solution channel by the culture solution import and culture solution outlet；
The gas being connected to the gas feed provides device, and the gas provides to be provided between device and the gas feed
Gas flow regulating valve；
The control system being electrically connected with the gas flow regulating valve；
The culture solution conveying device is used to convey culture solution to the culture solution channel, and the control system is described for controlling
The flow that the aperture of gas flow regulating valve is passed through gas into the gas dissolver to gas offer device is adjusted
Section；
The system also includes pressure extension box；Filled with gas in the pressure extension box, the bottom of the pressure extension box with
The culture solution channel or cell growth chamber connection, the control system are also used to control the culture solution conveying device
Culture solution is flowed through to the culture solution channel in the form of impulse wave, the pressure extension box is used to make by gas volume variation
Obtain the pressure balance in the culture solution channel and the cell growth chamber.
2. automatic cytological amplification system according to claim 1, which is characterized in that
Hollow fiber conduit is provided in the shell of the gas dissolver, the hollow fiber conduit separates space in the shell
Side and gas flow side are flowed through for culture solution, the culture solution flows through side and the culture solution import and culture solution outlet connects
Logical, the gas flow side is connected to the gas feed.
3. automatic cytological amplification system according to claim 1, which is characterized in that
It includes dioxide bottle and oxygen gas cylinder that the gas, which provides device, the dioxide bottle and the gas feed
Between be provided with first flow regulating valve, be provided with second flow regulating valve between the oxygen gas cylinder and the gas feed,
The control system is electrically connected with the first flow regulating valve and the second flow regulating valve, for being passed through the gas
The flow of carbon dioxide and oxygen in dissolvers is adjusted.
4. automatic cytological amplification system according to claim 3, which is characterized in that
The control system is used to obtain the consumption models of carbon dioxide and the consumption models of oxygen, and according to the carbon dioxide
Consumption models the aperture of the first flow regulating valve is adjusted so that the pH value of culture solution is maintained at the first default model
In enclosing, it is adjusted according to aperture of the consumption models of the oxygen to the second flow regulating valve, so that culture solution is molten
Oxygen amount is maintained in the second preset range.
5. automatic cytological amplification system according to claim 3, which is characterized in that
PH detector and dissolved oxygen amount detector, the control system and the pH detector and institute are provided on the circulation conduit
The electrical connection of dissolved oxygen amount detector is stated, the pH signal that the control system is used to send according to the pH detector is to described first-class
The aperture of adjustable valve is adjusted, so that pH value is maintained in the first preset range；It is sent according to the dissolved oxygen amount detector
Dissolved oxygen amount signal the aperture of the second flow regulating valve is adjusted so that dissolved oxygen amount is maintained at the second preset range
It is interior.
6. automatic cytological amplification system according to claim 1, which is characterized in that
The culture solution conveying device further includes circulating pump, the output end of the circulating pump and the culture solution inlet communication, defeated
Enter the outlet at end with the culture solution channel, the circulating pump is electrically connected with the control system.
7. automatic cytological amplification system according to claim 6, which is characterized in that
The cell growth chamber is provided with first pressure sensor, culture solution outlet and the culture solution of the gas dissolver
Second pressure sensor is additionally provided on circulation conduit between the import in channel, the control system and the first pressure pass
Sensor and second pressure sensor electrical connection, the control system are also used to obtain the metabolite of the cell growth chamber
Concentration model of growth, and followed according to the pressure control that the first pressure sensor and the second pressure sensor are sent
Ring pump conveys culture solution to the culture solution channel with predetermined pulse model, so that the Metabolites Concentration of the cell growth chamber
It is maintained in third preset range.
8. automatic cytological amplification system according to claim 6, which is characterized in that
The system also includes: waste liquid delivery pump and waste tank；
The culture solution conveying device further includes the culture solution being arranged between the hollow fiber reactor and the circulating pump
Surge tank, the outlet of the import of the culture solution surge tank and the culture solution channel are exported defeated with the circulating pump
Enter end connection；
Pipeline between the input terminal of the waste liquid delivery pump and the culture solution surge tank and the culture solution channel is connected to, defeated
Outlet is connected to the waste tank, and the control system is electrically connected with the waste liquid delivery pump；
The control system is also used to obtain the model of growth of the products of cellular metabolism in the culture solution channel, however, it is determined that cell generation
It thanks to the difference between the preset threshold of product and the concentration of the products of cellular metabolism in the culture solution channel and is less than or equal to first in advance
If when threshold value, then controlling the waste liquid delivery pump and opening, the culture solution in the culture solution channel is delivered in waste tank.
9. automatic cytological amplification system according to claim 1, which is characterized in that
The system also includes sealant supplement cylinders and culture solution delivery pump；
The cell growth chamber further includes import, and the outlet of the sealant supplement cylinder is connected to the input terminal of the culture solution delivery pump,
The inlet communication of the output end of the culture solution delivery pump and the cell growth chamber, the culture solution outlet of the gas dissolver
Temperature sensor is additionally provided on the circulation conduit being connected to the culture solution channel, the sealant supplement cylinder and the culture solution convey
It pumps and is additionally provided with heater, the control system and the culture solution delivery pump, temperature sensor and heating on the pipeline of connection
Device electrical connection；
The control system is also used to obtain the temperature signal that the temperature sensor is sent, and to the culture solution delivery pump and
The open and close of the heater are controlled, so that the temperature in the culture solution channel is maintained in the 4th preset range.
10. automatic cytological amplification system according to claim 9, which is characterized in that
It is additionally provided with third pressure sensor on the pipeline of the culture solution delivery pump and the inlet communication of the cell growth chamber,
The control system is electrically connected with the third pressure sensor, and the control system is also used to according to the third pressure sensing
The pressure signal that device is sent controls the speed of culture solution delivery pump conveying culture solution, so that the culture solution conveys
The speed of pump conveying culture solution is maintained in the 5th preset range.
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| 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 |
| CN107012094B (en) * | 2017-05-26 | 2019-03-22 | 奥凯（苏州）生物技术有限公司 | A kind of module group unit cell expansion system |
| CN107043734B (en) * | 2017-05-26 | 2020-12-25 | 奥凯（苏州）生物技术有限公司 | Automatic cell collection method of cell amplification system |
| CN107904167B (en) * | 2017-11-15 | 2023-06-06 | 中国航天员科研训练中心 | Automatic cell expansion culture equipment |
| CN107904170A (en) * | 2017-12-18 | 2018-04-13 | 上海白泽医疗器械有限公司 | Media to cell culture modules and cell culture system |
| CN108300661A (en) * | 2018-02-13 | 2018-07-20 | 京东方科技集团股份有限公司 | Culture apparatus and culture device |
| CN110404112B (en) * | 2018-04-26 | 2022-03-04 | 西安新长安再生医学科技研发有限公司 | Preparation method of self-body beautifying biological material and self-body beautifying biological material |
| US20190352589A1 (en) * | 2018-05-18 | 2019-11-21 | Huacells Corporation | Automated Cell Culturing |
| CN111334427B (en) * | 2019-03-12 | 2023-05-05 | 华道(上海)生物医药有限公司 | Totally-enclosed cell culture system |
| CN109913410A (en) * | 2019-04-19 | 2019-06-21 | 华子昂 | The emulation cultural method of stem cell |
| WO2021114129A1 (en) * | 2019-12-11 | 2021-06-17 | 中国科学院深圳先进技术研究院 | In vitro experimental model for research of mechanism of tumor perineural invasion |
| CN112940933A (en) * | 2021-01-29 | 2021-06-11 | 上海睿钰生物科技有限公司 | In-vitro life-sustaining perfusion culture system and control method thereof |
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| CN1131304C (en) * | 1999-12-09 | 2003-12-17 | 中国科学院力学研究所 | Spacial dialyzing counterflow cell cultivator and cultivating method |
| CN101486967B (en) * | 2009-02-13 | 2011-09-28 | 中国人民解放军第三军医大学第一附属医院 | Culture fluid gas/liquid and liquid/liquid exchanger for tissue engineering tissue bionic cultivation |
| 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 |
| 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 |
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