CN203923208U - The supporting cell culture system of disposable sterilized cell culture bags - Google Patents

The supporting cell culture system of disposable sterilized cell culture bags Download PDF

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Publication number
CN203923208U
CN203923208U CN201420293029.9U CN201420293029U CN203923208U CN 203923208 U CN203923208 U CN 203923208U CN 201420293029 U CN201420293029 U CN 201420293029U CN 203923208 U CN203923208 U CN 203923208U
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cell culture
unit
control unit
gas
disposable sterilized
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CN201420293029.9U
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何国强
陈跃武
邓哲
刘利科
梁志国
胡志威
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AUSTAR PHARMACEUTICAL EQUIPMENT (SHIJIAZHUANG) Co Ltd
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AUSTAR PHARMACEUTICAL EQUIPMENT (SHIJIAZHUANG) Co Ltd
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Abstract

The utility model provides a kind of disposable sterilized cell culture bags supporting cell culture system, comprise agitating unit, support pipeline, temperature control unit, the controller unit of gas circuit control unit and control four unit noted earlier, described support pipeline respectively with agitating unit, temperature control unit, gas circuit control unit is connected, in order to provide described system required gas, the passage of liquid, described agitating unit is provided with the agitating vane with the integrated design of hollow gas distributor, described hollow gas distributor is connected with gas circuit control unit, described hollow gas distributor comprises the micro bubbles sparger that is arranged on agitating vane bottom and the ventpipe that is arranged on agitating vane inside.Described system has reduced labour intensity, produces required personnel and GMP production plant area will be significantly less than traditional production unit, is conducive to realize the operation of reequiping faster for use in new technological process.

Description

The supporting cell culture system of disposable sterilized cell culture bags
Technical field
The utility model relates to a kind of cell culture system, especially relates to the supporting cell culture system of a kind of disposable sterilized cell culture bags.
Background technology
Large scale culturing technology (large-scale culture technology) refers under artificial condition (sets ph, temperature, dissolved oxygen etc.), the technology at a large amount of culturing cells of bioreactor (bioreactor) middle-high density for the production of biological products.
Large scale culturing cell mainly adopts spinner culture system and stainless cylinder of steel culture systems at present.
There is following problem in spinner culture: its surface-area is limited, and Growth of Cells density is low; When cultivation, monitoring and the condition that controls environment (pH, dissolved oxygen etc.) are unstable, affect quality product; Labour intensity is large, takes up room large, and pollution rate is high.
Stainless cylinder of steel is cultivated the following problem that exists: need complicated cleaning in place (CIP) or sterilizing in place (SIP) pipeline, the possibility of crossed contamination between technical process operation; Every production batch setup time is long, and producers' labour intensity is large; Stirring system shearing force is large, and cell injury is serious.
Utility model content
The utility model provides a kind of disposable sterilized cell culture bags supporting cell culture system, can meet well the needs of cell height propagation, coordinate scientific and reasonable online monitoring system to reflect in time the upgrowth situation of cell simultaneously, make cell high-density culture more scientific and reasonable, ensure the high efficient expression of target product.Its technical scheme is as described below:
The supporting cell culture system of disposable sterilized cell culture bags, comprise agitating unit, support pipeline, temperature control unit, the controller unit of gas circuit control unit and control four unit noted earlier, described support pipeline respectively with agitating unit, temperature control unit, gas circuit control unit is connected, in order to provide described system required gas, the passage of liquid, described agitating unit is provided with the agitating vane with the integrated design of hollow gas distributor, described hollow gas distributor is connected with gas circuit control unit, described hollow gas distributor comprises the micro bubbles sparger that is arranged on agitating vane bottom and the ventpipe that is arranged on agitating vane inside.
Described controller unit is provided with the touch-screen that carries out man-machine interaction, and Control Server, and by PLC, other four unit is carried out to automatization control.
The top of described agitating unit is provided with the motor that drives agitating vane, and described motor adopts Frequency Converter Control.
Described support pipeline is provided with the valve of the opening and closing to gas, liquid line, described support pipeline is divided into gas piping system and process pipeline system, and described gas piping system comprises that pressurized air, oxygen, nitrogen, the carbon dioxide mouth of pipe, flow take into account by-pass valve control; Described process pipeline system comprises alkali pump, product pump, temperature control water.
Described cell culture bags is placed in jacketed vessel, described jacketed vessel is connected by high-pressure water pipe, rapid-acting coupling with temperature control unit, and described temperature control unit comprises the recycle pump that the water in chuck is circulated, the refrigerator that water temperature is controlled and well heater.
Described gas circuit control unit is provided with pH sensor and dissolved oxygen sensor, for measuring pH value and the oxygen dissolving value of cell culture fluid.
In described jacketed vessel, be provided with the temperature sensor being connected with controller unit, described controller unit is connected with recycle pump, refrigerator and well heater.
Described pH sensor is connected with controller unit with dissolved oxygen sensor, pressurized air, O in described controller unit control support pipeline 2, N 2and CO 2the open and close of AVSV Air Vent Solenoid Valve.
In described cell culture bags, be provided with the pressure transmitter and the temperature sensor that are connected with controller unit.
In the pipeline of described gas piping system, be provided with the under meter being connected with controller unit.
The supporting cell culture system of disposable sterilized cell culture bags that the utility model provides, it is the complete Controlling System designing for disposable cell bags culture scheme, especially stirring rake and gas distributor integrated design, the gaseous conductance speed of ventilation capacity and oxygen is high, there is extremely low mechanical shear stress simultaneously, described system, because adopting disposable assembly, has been avoided the possibility of crossed contamination between technical process operation; Reduce every production batch setup time.
Brief description of the drawings
Fig. 1 is the supporting cell culture system framework of disposable sterilized cell culture bags described in the utility model schematic diagram;
Fig. 2 is the structural representation of the supporting cell culture system of described disposable sterilized cell culture bags;
Fig. 3 is the external structure schematic diagram of the supporting cell culture system of described disposable sterilized cell culture bags;
Fig. 4 is the side-view of Fig. 3;
Fig. 5 is A-A sectional view in Fig. 4;
Fig. 6 is the schematic diagram of stirring rod position;
Fig. 7 is the schematic diagram of described agitating vane;
Fig. 8 is the schematic diagram of described hollow gas distributor;
Fig. 9 is the side-view of described hollow gas distributor;
1-electrical cabinet framework, 2-pedestal, 3-stirring-head, 4-regulator cubicle, 5-universal wheel, 6-directional wheel, 7-chain female connector, supports in 8-stirring rod, 9-stirring rod lower support, 10-stirring rod (25L uses), 11-stirring rod (50L uses), 12-foam bag support, 13-stirs head-shield, 14-protective door, 15-container guide rail, 16-agitating vane, 17-hollow gas distributor, 18-ventpipe, 19-micro bubbles sparger, 20-sleeve.
Embodiment
As shown in Figure 1, concrete structure schematic diagram as shown in Figure 2, is mainly made up of parts such as agitating unit, support pipeline, temperature control unit, gas circuit control unit and controllers the supporting cell culture system of disposable sterilized cell culture bags that the utility model provides.By support pipeline, agitating unit, temperature control unit, gas circuit control unit are coupled together; Controller unit carries out automatization control by PLC to above four unit, and carries out man-machine interaction by touch-screen, can realize the automatic control functions such as culture bag mixing speed, temperature, pH, dissolved oxygen, and have data logging and query function.
As shown in Figure 3 and Figure 4, the supporting cell culture system of disposable sterilized cell culture bags is outside equipped with electrical cabinet framework 1, described electrical cabinet framework 1 is placed on pedestal 2, described base 2 is provided with container guide rail 15, universal wheel 5, directional wheel 6 and chain female connector 7, and described directional wheel 6 can be seen in Fig. 6.
On described electrical cabinet framework 1, be provided with and stir head-shield 13, protective door 14 and support 12 for foam bag, in order to immobilized-cell culture bag.
As shown in Figure 5, described stirring-head 3 is arranged on described electrical cabinet framework 1, belongs to agitating unit.Comprise motor, step-down gear, drive socket, after stirring rod upper end is inserted in drive socket, insert in agitating vane 16 stirring rod lower end, and revolution, can drive blade to stir.
In Fig. 6, in the stirring rod arranging in described electrical cabinet framework 1, support 8, stirring rod lower support 9 is connected with respectively 25L stirring rod 10 and stirring rod 11 for 50L.
According to the difference of cell bags volume (25L or 50L) and support vessels, system is furnished with 2 different stirring rod.Stirring rod by supporting 8 in stirring rod, stirring rod lower support 9 is fixed on Electric Appliance Cabinet framework 1.
As shown in Fig. 7, Fig. 8, Fig. 9, described hollow gas distributor 17 comprises the micro bubbles sparger 19 that is arranged on agitating vane 16 bottoms and the ventpipe 18 that is arranged on agitating vane inside, described ventpipe 18 is connected with gas circuit control unit with micro bubbles sparger 19 respectively, the gas of transmission gas circuit control unit is to micro bubbles sparger 19, described micro bubbles sparger 19 is made up of miniature air outlet one by one, can effectively see through gas, and prevent that liquid from entering ventpipe.
Agitating vane 16, ventpipe 18 and micro bubbles sparger 19 are all fixed on sleeve 20, described sleeve 20 is that stirring rod is connected use with agitating vane 16, in the time that stirring rod drives agitating vane 16 to move, ventpipe 18 and micro bubbles sparger 19 be operation thereupon also, makes diffusing in nutrient solution of gas uniform.
Illustrate the effect of each unit below in conjunction with Fig. 2.
The top of described agitating unit is provided with motor, drives agitating vane to stir, and drives flowing of feed liquid, realizes and mixing and mass transfer effect.Motor adopts Frequency Converter Control, and stepless change, operates steadily, and mixing speed is adjustable within the scope of (20~120) rpm.
Described frequency transformer is accepted analog quantity setting signal, the digital quantity start stop signal from controller unit by quick connector, and current frequency transformer output speed, run signal, trouble signal are fed back to controller unit.Agitating unit simultaneously with cell culture bags in pressure, temperature carry out interlocking alarm, when bag internal pressure or temperature exceed alarm setting value, stir and will stop in emergency.
The vane type adopting with respect to stainless cylinder of steel stirs, stirring rake realize shearing force extremely low in, chemistry (nutrition and waste), physics (pH, an O uniformly can be provided in tank body 2and CO 2) and heating power environment, and maintain being uniformly distributed of cell suspension.
Stirring rake 16 and 17 integrated designs of hollow gas distributor, this design forms relatively high surface volume ratio via the bubble producing by hollow gas distributor and headspace, has improved the gaseous conductance speed of nutrient solution and oxygen; Pass into gas in nutrient solution time, stir the effect that can produce DYNAMIC DISTRIBUTION, also can promote oxygen conduction and CO 2release.
Described support pipeline provides gas, and the passage of liquid carries out the opening and closing control of pipeline by various valves.By the monitoring of the device such as well heater, refrigerator, peristaltic pump, gas filter, gas meter, gas ratio variable valve, maintain the stable of parameter in reactor operational process.
It is divided into gas piping system and process pipeline system according to function, and gas piping system comprises that pressurized air, oxygen, nitrogen, the carbon dioxide mouth of pipe, flow take into account by-pass valve control etc.; Process pipeline system comprises respectively alkali pump, product pump, temperature control water etc.Between pipeline, connect by aseptic rapid-acting coupling.
In described temperature control unit, cell bags is placed in jacketed vessel, and chuck is connected by high-pressure water pipe, rapid-acting coupling with temperature control unit.Recycle pump circulates to the water in chuck, and refrigerator and well heater are controlled water temperature.Chuck and cell bags are carried out heat exchange and realize the temperature control of nutrient solution in final cell bag.
Principle of work is as described below: by adopting 2 temperature electrodes to measure cell bags temperature and jacket temperature, by rapid-acting coupling, signal is transferred to control unit, control unit is pressed two temperature pid control mode according to set(ting)value and process values, controls refrigerator and well heater and opens/stop and realize temperature control.When cell bags design temperature is during lower than the probe temperature temperature difference, system starts heating automatically; When design temperature is during higher than probe temperature, system starts refrigeration automatically.Reasonably control strategy and parameter setting have ensured the temperature controlled susceptibility of cell bags and stability.
Therefore, temperature of reactor control unit adopts the two temperature pid control mode based on cell bags temperature, jacket temperature, temperature control precision ± 0.2 DEG C.
The various gases that described gas circuit control unit need to provide cell to need, oxygen is that cellular metabolism is necessary.Although cell cultures is aerobic far below microbial culture, oxygenate is still a major challenge that cell cultures scale is amplified.Air flow need to improve with the increase of biological reaction container (being cell bags) volume and cell concn herein.Oxygen level reduces will affect Growth of Cells and output, and higher oxygen level also can produce toxicity to cell.
Air, O 2, CO 2and the N using once in a while 2inject cell culture fluid by hollow gas distributor.Minitype gas sparger can provide larger gas-to-liquid contact surface, and higher oxygen mass transfer coefficient (kLa) therefore can be provided under lower gas flow rate.
Wherein, pH control method is to adopt pH electrode to measure, and control unit is realized according to pid algorithm control carbonic acid gas breather valve and alkali lye peristaltic pump.When pH is during lower than set(ting)value, control unit starts alkali lye peristaltic pump, supplements alkali lye and improves pH to set(ting)value; When pH is during higher than set(ting)value, control unit starts carbon dioxide magnetic valve, reduces pH to set(ting)value.Alkali lye peristaltic pump flow can regulate by peristaltic pump motor driver, has ensured the sensitivity that pH controls.
Dissolved oxygen control method is to adopt dissolved oxygen electrode to measure, and control unit is according to senior pid algorithm control air, O 2, N 2the opening time of magnetic valve is realized the automatic control of dissolved oxygen.When dissolved oxygen is during lower than set(ting)value, controller starts control valve, improves dissolved oxygen to set(ting)value to logical oxygen/pressurized air in reactor.When dissolved oxygen is during higher than set(ting)value, controller starts control valve, maintains dissolved oxygen to set(ting)value to logical appropriate nitrogen in reactor.
This culture systems assembled two of DO (being dissolved oxygen) and pH independently, the air-flow control strategy of applying flexible, to automatically maintain DO and pH setup parameter.In these two strategies, nearly four kinds of gases (air, O 2, N 2and CO 2) can automatically regulate correct blending ratio by the feedback of DO and pH sensor.Then, mixed gas is input in gas distributor by a thermal mass gas flow meters (TMFC).PLC in control unit controls air, O 2, N 2and CO 2the On/Off of AVSV Air Vent Solenoid Valve; To TMFC, and accept the feedback signal of TMFC by rapid-acting coupling transportation simulator amount setting signal; The opening/stop of output high-speed pulse control alkali lye peristaltic pump by PLC.
Described controller unit is taking PLC and module thereof as core, in PLC according to processing requirement coding, collocation touch-screen, dissolved oxygen transmitter, pH transmitter, peristaltic pump, magnetic valve etc. are realized the control function of whole system, various state parameters while gathering cell cultures, and carry out automatic or manual control.
Controller unit adopts touch-screen control and two kinds of patterns of terminal Long-distance Control.
Show the critical datas such as DO, pH, temperature, mixing speed by touch-screen, and the current running status such as valve, pump, motor.And can carry out the unlatching of the elements such as valve, pump, electrical equipment on touch-screen, and feature operation and parameter setting (comprising that the setting at Temperature Setting, DO/pH reference mark, PID set).There is data presentation, state of the art demonstration, process parameter control, correction, the functions such as data logging storage, the inquiry of historical data, meet GMP and manage record and the trackability requirement to goods.
When terminal Long-distance Control, operator can realize operation and the monitoring to culture systems by terminal computer.
Described cell culture system adopts disposable sterilized cell culture bags, it when arrival, is independent sterile packed, penetrate sterilization via gamma ray good fortune, instant getting, setup time between having reduced batch, reduce labour intensity, produced required personnel and GMP production plant area will be significantly less than traditional production unit.
Disposable sterilized cell culture bags and assembly thereof are not used in follow-up operation, have eliminated the possibility of crossed contamination between the common technical process operation of traditional tank body.Because reduce or abandoned the demand to stainless steel equipment, also can avoid the long period of device assembles.The reduced complexity of system, thereby related engineering demand reduces too.Without cleaning in place (CIP) or sterilizing in place (SIP) operation, and the pressure rating of relevant pipeline, valve, controller or container.In addition, the use of disposable assembly has also reduced the complexity of checking.Because almost there is no repeatedly used assembly, just do not have any project to need to follow the tracks of yet, also just save a large amount of sterilizings, clean checking research.Finally, by having eliminated the restriction of firm tubing system and fixing stainless cylinder of steel, disposable assembly also helps realization and reequips faster the operation for use in new technological process.
The stirring rake system of particular design has extremely low mechanical shear stress, can greatly lower cell injury or mortality ratio.Stirring rake and the integrated design of hollow gas distributor, produce the effect of DYNAMIC DISTRIBUTION in nutrient solution while passing into gas, improve the gaseous conductance speed of ventilation capacity and oxygen.
For disposable cell culture bags, set up rational control model, design brand-new, reliable and stable control strategy, carry out the optimum control of pH and dissolved oxygen (DO).Culture systems can be controlled the flow that adds air, oxygen, nitrogen and four kinds of gases of carbonic acid gas in cell culture bags in an orderly manner quantitatively by controller, make it keep best ratio to control pH value and the dissolved oxygen level in cell culture fluid, make system all the time in optimum regime, the needs with the growth that meets zooblast to pH value and dissolved oxygen.

Claims (10)

1. the supporting cell culture system of disposable sterilized cell culture bags, it is characterized in that: comprise agitating unit, support pipeline, temperature control unit, the controller unit of gas circuit control unit and control four unit noted earlier, described support pipeline respectively with agitating unit, temperature control unit, gas circuit control unit is connected, in order to provide described system required gas, the passage of liquid, described agitating unit is provided with the agitating vane with the integrated design of hollow gas distributor, described hollow gas distributor is connected with gas circuit control unit, described hollow gas distributor comprises the micro bubbles sparger that is arranged on agitating vane bottom and the ventpipe that is arranged on agitating vane inside.
2. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, it is characterized in that: described controller unit is provided with the touch-screen that carries out man-machine interaction, and Control Server, and by PLC, other four unit are carried out to automatization control.
3. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, is characterized in that: the top of described agitating unit is provided with the motor that drives agitating vane, and described motor adopts Frequency Converter Control.
4. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, it is characterized in that: described support pipeline is provided with the valve of the opening and closing to gas, liquid line, described support pipeline is divided into gas piping system and process pipeline system, and described gas piping system comprises that pressurized air, oxygen, nitrogen, the carbon dioxide mouth of pipe, flow take into account by-pass valve control; Described process pipeline system comprises alkali pump, product pump, temperature control water.
5. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, it is characterized in that: described cell culture bags is placed in jacketed vessel, described jacketed vessel is connected by high-pressure water pipe, rapid-acting coupling with temperature control unit, and described temperature control unit comprises the recycle pump that the water in chuck is circulated, the refrigerator that water temperature is controlled and well heater.
6. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, is characterized in that: described gas circuit control unit is provided with pH sensor and dissolved oxygen sensor, for measuring pH value and the oxygen dissolving value of cell culture fluid.
7. the supporting cell culture system of disposable sterilized cell culture bags according to claim 5, it is characterized in that: in described jacketed vessel, be provided with the temperature sensor being connected with controller unit, described controller unit is connected with recycle pump, refrigerator and well heater.
8. the supporting cell culture system of disposable sterilized cell culture bags according to claim 6, is characterized in that: described pH sensor is connected with controller unit with dissolved oxygen sensor, pressurized air, O in described controller unit control support pipeline 2, N 2and CO 2the open and close of AVSV Air Vent Solenoid Valve.
9. the supporting cell culture system of disposable sterilized cell culture bags according to claim 1, is characterized in that: in described cell culture bags, be provided with the pressure transmitter and the temperature sensor that are connected with controller unit.
10. the supporting cell culture system of disposable sterilized cell culture bags according to claim 4, is characterized in that: in the pipeline of described gas piping system, be provided with the under meter being connected with controller unit.
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CN104017727A (en) * 2014-06-04 2014-09-03 奥星制药设备(石家庄)有限公司 Disposable supporting cell culture system for sterile cell culture bag
CN105420104A (en) * 2015-12-09 2016-03-23 中国航天员科研训练中心 Cell culture bag with three-dimensional structure
US9536122B2 (en) 2014-11-04 2017-01-03 General Electric Company Disposable multivariable sensing devices having radio frequency based sensors
US9538657B2 (en) 2012-06-29 2017-01-03 General Electric Company Resonant sensor and an associated sensing method
US9638653B2 (en) 2010-11-09 2017-05-02 General Electricity Company Highly selective chemical and biological sensors
US9658178B2 (en) 2012-09-28 2017-05-23 General Electric Company Sensor systems for measuring an interface level in a multi-phase fluid composition
CN106802684A (en) * 2016-12-30 2017-06-06 徐州工程学院 Purple potato pears compound fruit vinegar fermentation monitoring system and its method of work based on MAS
US9746452B2 (en) 2012-08-22 2017-08-29 General Electric Company Wireless system and method for measuring an operative condition of a machine
US9920292B2 (en) 2015-08-31 2018-03-20 General Electric Company System and method for initiating a cell culture
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US10684268B2 (en) 2012-09-28 2020-06-16 Bl Technologies, Inc. Sensor systems for measuring an interface level in a multi-phase fluid composition
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US10018613B2 (en) 2006-11-16 2018-07-10 General Electric Company Sensing system and method for analyzing a fluid at an industrial site
US9638653B2 (en) 2010-11-09 2017-05-02 General Electricity Company Highly selective chemical and biological sensors
US9538657B2 (en) 2012-06-29 2017-01-03 General Electric Company Resonant sensor and an associated sensing method
US10598650B2 (en) 2012-08-22 2020-03-24 General Electric Company System and method for measuring an operative condition of a machine
US9746452B2 (en) 2012-08-22 2017-08-29 General Electric Company Wireless system and method for measuring an operative condition of a machine
US9658178B2 (en) 2012-09-28 2017-05-23 General Electric Company Sensor systems for measuring an interface level in a multi-phase fluid composition
US10684268B2 (en) 2012-09-28 2020-06-16 Bl Technologies, Inc. Sensor systems for measuring an interface level in a multi-phase fluid composition
CN104017727A (en) * 2014-06-04 2014-09-03 奥星制药设备(石家庄)有限公司 Disposable supporting cell culture system for sterile cell culture bag
CN104017727B (en) * 2014-06-04 2016-03-30 奥星制药设备(石家庄)有限公司 The supporting cell culture system of disposable sterilized cell culture bags
US9536122B2 (en) 2014-11-04 2017-01-03 General Electric Company Disposable multivariable sensing devices having radio frequency based sensors
US9920292B2 (en) 2015-08-31 2018-03-20 General Electric Company System and method for initiating a cell culture
CN105420104A (en) * 2015-12-09 2016-03-23 中国航天员科研训练中心 Cell culture bag with three-dimensional structure
CN106802684A (en) * 2016-12-30 2017-06-06 徐州工程学院 Purple potato pears compound fruit vinegar fermentation monitoring system and its method of work based on MAS

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