KR102473972B1 - Single use cell culture system - Google Patents

Abstract

The present invention provides a disposable cell culture system, comprising: a biological safety cabinet (110) having a support (111) formed at a set height from the ground and a plate (112) disposed on the top of the support (111) and configured to open and close a portion of the front surface or a portion of the side surface; a wave bioreactor (120) having a seating table (121) formed on the top of the plate (112) to seat a disposable bag (10) thereon and an agitator (122) agitating a cell culture solution of the disposable bag (10) by vibrating the seating table (121) up and down or left and right; a gas mixer controller (130) formed on one side of the biological safety cabinet (110) to control gas supply to the disposable bag (10); and a measurement unit (140) formed on the seating table (121) to be connected to each sensor port of the disposable bag (10) so that pH, DO, temperature, and foam are measured; and a bioreactor controller (150) formed on the other side of the biological safety cabinet (110) to control the wave bioreactor (120), calibrating each sensor of the measurement unit (140), programming the culture environment of the cell culture solution, recording and maintaining the fermentation process of the cell culture solution, controlling the agitation speed of the agitator (122), and enabling control of DO, thereby operating the wave bioreactor (120) in the internal space of the biological safety cabinet (110) to improve safety and perform long-time fermentation in an aseptic environment.

Description

Disposable cell culture system {SINGLE USE CELL CULTURE SYSTEM}

The present invention relates to a disposable cell culture system capable of improving safety by operating a wave bioreactor in an internal space of a biosafety cabinet and fermenting a cell culture solution in a disposable bag in an aseptic environment for a long time.

As is well known, various types of bioreactors including cell culture devices are widely used in a wide range of fields such as medicine, pharmacy, and biotechnology, such as reagent and vaccine manufacturing, new drug development, treatment and research using stem cells.

For example, various conditions such as nutrients, temperature, humidity, pH, concentration of oxygen and carbon dioxide suitable for the inside of the container where the mixing of various cell culture media or cell culture in a bioreactor occur must be maintained for a specific period of time. do.

On the other hand, if the culture medium being produced using the bioreactor is contaminated, all expensive materials in use must be discarded. Therefore, the inside of the bioreactor must be kept completely shielded from the outside so as not to be contaminated.

Accordingly, a technology capable of fermenting the cell culture solution in a disposable bag for a long time in an aseptic environment through a biological safety cabinet is required.

Korean Patent Registration No. 10-2076363 (disposable biological fermenter) Korean Patent Registration No. 10-1629150 (Agitator for bioreactor using a disposable container)

The technical problem to be achieved by the spirit of the present invention is to improve safety by operating a wave bioreactor in the inner space of a biological safety cabinet and to provide a disposable cell culture system that can ferment a cell culture medium in a disposable bag for a long time in an aseptic environment. have.

In order to achieve the above object, an embodiment of the present invention is a biosafety cabinet having a support formed at a certain height from the ground, a plate disposed on top of the support, and configured to open or close a part of the front or part of the side; a wave bioreactor having a seat formed on the top of the plate on which the disposable bag is seated, and a stirrer configured to agitate the cell culture medium of the disposable bag by vibrating the seat up and down or left and right; a gas mixer controller formed on one side of the biosafety cabinet to control gas supply to the disposable bag; a measurement unit formed on the safety belt and connected to each sensor port of the disposable bag to measure pH, DO, temperature, and foam, respectively; And formed on the other side of the biological safety cabinet to control the wave bioreactor, calibrate each sensor of the measurement unit, program the culture environment of the cell culture medium, record and maintain the fermentation process of the cell culture medium, It provides a disposable cell culture system comprising a; bioreactor controller that controls the stirring speed of the stirrer and enables control of DO.

Here, an inlet duct that guides outside air taken in through a partially opened gap of the biosafety cabinet to the top of the biosafety cabinet through the rear surface, an intake fan that takes in outdoor air through the inlet duct, and passing through the intake fan It may further include an air circulation unit composed of a filter for filtering air and a circulation duct for circulating the air passing through the filter in the inner space of the biological safety cabinet.

In addition, batch, fed-batch and continuous fermentation can be performed through the bioreactor controller.

In addition, substitution of carbon dioxide and nitrogen may be possible through the gas mixer controller.

In addition, a disposable vessel may be seated on the plate by replacing the wave bioreactor, and the gas mixer controller may control gas supply to the disposable vessel.

In addition, the disposable vessel includes a rotating shaft formed on a central axis, a turbine impeller formed radially symmetrical at a lower end of the rotating shaft, and a top magnetic drive formed at a lower end of the turbine impeller, and a top magnetic drive formed at a lower end of the plate. A lower magnetic drive opposite to the drive and a servo motor rotationally driving the lower magnetic drive may be formed.

According to the present invention, there is an effect of improving safety by operating the wave bioreactor in the inner space of the biological safety cabinet and fermenting the cell culture solution in a disposable bag for a long time in an aseptic environment.

1 is an implementation diagram of a disposable cell culture system according to an embodiment of the present invention.
Figure 2 shows a front view of the disposable cell culture system of Figure 1.
Figure 3 is a separate view of the wave bioreactor of the disposable cell culture system of Figure 1.
FIG. 4 illustrates air circulation of the disposable cell culture system of FIG. 1 .
FIG. 5 illustrates the application of the disposable bag of the wave bioreactor of FIG. 1 .
Figure 6 illustrates the application of the disposable vessel of Figure 1.
FIG. 7 illustrates a perspective view of the disposable cell culture system of FIG. 1 .
8 illustrates an exploded view of the disposable cell culture system of FIG. 7 .
FIG. 9 illustrates a cross-sectional structure of the disposable cell culture system of FIG. 7 .

Hereinafter, embodiments of the present invention having the above-described characteristics will be described in more detail with reference to the accompanying drawings.

The disposable cell culture system according to an embodiment of the present invention includes a support 111 formed at a certain height from the ground, and a plate 112 disposed on top of the support 111, so that a part of the front side or a part of the side is opened and closed. The biosafety cabinet 110 formed on the top of the plate 112 and the seating table 121 on which the disposable bag 10 is seated, and the seating table 121 is vibrated up and down or left and right to vibrate the cells of the disposable bag 10 A wave bioreactor 120 having an agitator 122 for agitating the culture medium, a gas mixer controller 130 formed on one side of the biosafety cabinet 110 and controlling gas supply to the disposable bag 10, and seating It is formed on the table 121 and is connected to each sensor port of the disposable bag 10 to measure pH, DO, temperature and foam, respectively. Control the reactor 120, calibrate each sensor of the measuring unit 140, program the culture environment of the cell culture medium, record and maintain the fermentation process of the cell culture medium, control the stirring speed of the stirrer 122, Including the bioreactor controller 150 that enables the control of DO, the wave bioreactor 120 is operated in the internal space of the biological safety cabinet 110 to improve safety and to ferment for a long time in an aseptic environment. do.

Hereinafter, with reference to FIGS. 1 to 9, the detailed description of the disposable cell culture system having the above configuration is as follows.

First of all, the biosafety cabinet 110 is a configuration that provides an internal space in an aseptic environment to enable a long-term culture process. Specifically, the support 111 formed at a certain height from the ground, and placed on top of the support 111 A plate 112 is provided, and a part of the front side or part of the side is configured to be opened and closed so that outside air is sucked in or the single use bag 10 is seated and separated.

Next, referring to FIGS. 1 to 3 , the wave bioreactor 120 includes a seating table 121 formed on top of the plate 112 and receiving the disposable bag 10 thereon, and a seating table 121 is provided with an agitator 122 for agitating the cell culture solution in the disposable bag 10 by vibrating up and down or left and right at regular intervals to provide an environment in which cells can grow and ferment the cell culture solution.

On the other hand, referring to FIG. 8, the seat 121 prevents the disposable bag 10 accommodated inside through a transparent cover 121a that can be opened and closed separately or hingedly, and the stirrer 122 is A plate 122a coupled to the bottom surface of 121, a cam link 122b swinging the plate 122a back and forth or left and right, a drive link 122c interlocked with the cam link 122b, and a drive link ( 122c) may be configured as a motor (122d) that rotates.

Here, although not shown, the seating table 121 may vibrate up and down or left and right by the agitator 122, but is rotated forward and backward at regular intervals on the horizontal surface of the seating table 121 to rotate the cells of the disposable bag 10. It may also be configured to agitate the culture medium.

Next, as illustrated in FIG. 5 , the gas mixer controller 130 is formed on one side of the rear side of the biosafety cabinet 110 and is connected to each port of the disposable bag 10 through a hose so that the disposable bag 10 Controls the supply of gases such as oxygen, carbon dioxide, and nitrogen to

Here, carbon dioxide and nitrogen can be substituted through the gas mixer controller 130, and nitrogen concentration and oxygen concentration can be controlled.

For example, the gas mixer controller 130 supplies a plurality of gas supply ports for supplying or removing various gases such as oxygen, carbon dioxide, nitrogen, and nitrogen to provide an anaerobic microorganism culture environment inside the disposable bag 10. A nitrogen sparger, an air supply port for supplying compressed air by a compressor, an inoculation source for supplying microorganisms, and a sampling port for collecting a culture solution sample of the cultured microorganism culture medium and supplying it to the sampler from the disposable bag 10, A feed pump for pumping corresponding gas to a gas supply port and a nitrogen sparger may be included, so that carbon dioxide and nitrogen may be substituted and nitrogen and oxygen concentrations may be controlled through valve control.

Next, the measuring unit 140 is formed on the seating table 121 and is connected to each sensor port (not shown) for pH, DO, and temperature of the disposable bag 10 to measure pH, DO, temperature, and foam, respectively.

Next, the bioreactor controller 150 is formed on the other side of the rear surface of the biological safety cabinet 110 to control the driving of the wave bioreactor 120, to calibrate each sensor of the measuring unit 140, and to culture the cell culture medium. is programmed, the fermentation process of the cell culture medium is recorded and maintained, the agitation speed of the stirrer 122 is controlled, and DO is controlled so as to control the fermentation process of the cell culture medium.

Here, through the bioreactor controller 150, various fermentation processes such as batch, fed-batch, and continuous fermentation of the cell culture solution can be selectively performed.

In addition, the bioreactor controller 150 may control the temperature of the cell culture medium in the disposable bag 10 through a warm air fan (not shown) or a hot wire (not shown) installed in the plate 112 .

Next, referring to FIG. 4 , the air circulation unit 150 guides outside air taken in through a partially opened gap on the front side of the biological safety cabinet 110 to the top of the biological safety cabinet 110 through the rear space. an inlet duct 151 that takes in air through the inlet duct 151, an intake fan 152 that takes in outside air, a filter 153 that filters foreign substances by filtering the air that has passed through the intake fan 152, and a filter 153 It consists of a circulation duct 154 that circulates the air that has passed through the biological safety cabinet 110 in the inner space of the biological safety cabinet 110, circulates the filtered air inside the biological safety cabinet 110, maintains an aseptic state, and blocks contamination. , It is possible to prevent cross-contamination between culture materials, and to prevent contaminants from leaking out of the biological safety cabinet 110 by intake of outdoor air, thereby protecting the experimenter and the laboratory.

Through this, through the aseptic environment of the biological safety cabinet 110, contamination of the culture during long-term cultivation is prevented, and contamination during inoculation and sampling by connecting the inoculation port of the small fermentor, the disposable bag 10, or the disposable vessel 20 can be blocked.

On the other hand, as illustrated in FIG. 6, in order to ferment the cell culture medium of the disposable vessel 20 instead of the cell culture medium of the disposable bag 10, the wave bioreactor 120 is replaced and placed on the plate 112. The disposable vessel 20 of the cylindrical transparent container is seated, and the gas mixer controller 130 may control gas supply to the disposable vessel 20 .

In addition, the disposable vessel 20 has a rotating shaft 161 formed on a central axis, a turbine impeller 162 formed radially symmetrical at the lower end of the rotating shaft 161, and a top magnetic drive formed at the lower end of the turbine impeller 162. 163, and at the lower end of the plate 112, a lower magnetic drive (not shown) opposite to the upper magnetic drive 163 and a servo motor (not shown) for rotationally driving the lower magnetic drive are formed, The impeller 162 may be rotated to agitate the cell culture medium of the disposable vessel 20 in a watertight state.

Meanwhile, before the disposable bag 10 or the disposable vessel 20 is seated on the plate 112, the internal space of the biological safety cabinet 110 may be sterilized by using high-temperature and high-pressure autoclaves or gamma rays of a gamma ray irradiator. .

In addition, through a PC connected to the bioreactor controller 150 by wire or wirelessly, temperature, agitation speed, pH, dissolved oxygen, and bubbles generated during fermentation can be easily and precisely controlled to cultivate anaerobic microorganisms, and through the PC , It is possible to integrate and control a plurality of bioreactor controllers 150 to enable data logging and remote control, and the bioreactor controller 150 enables data storage using a USB port, and the biological safety cabinet 110 ) can be configured to accommodate a wide range of cell culture tank capacities of 1.5 ml to 15 l, to facilitate cleaning after completion of culture by the biological safety cabinet 110, and to enable various culture experiments in a small laboratory It can be configured in a compact design.

Meanwhile, as illustrated in FIG. 9 , a spray nozzle for forming an air curtain is formed at the bottom of the front cover 113 or the bottom of the side cover 114 of the biosafety cabinet 110 to inoculate the disposable bag 10. At the time of inoculation and sampling by connection, compressed air is injected to the bottom to block contamination of the disposable bag 10 due to inflow of foreign substances from the outside, or to block air leakage from the seating table 121 to the outside It can also be augmented to perform inoculation or sampling operations in a clean environment.

Therefore, by the configuration of the disposable cell culture system as described above, the wave bioreactor can be operated in the inner space of the biological safety cabinet to improve safety and ferment the cell culture solution in the disposable bag for a long time in an aseptic environment.

The embodiments described in this specification and the configurations shown in the drawings are only one of the most preferred embodiments of the present invention, and do not represent all of the technical spirit of the present invention, so various equivalents that can replace them at the time of this application. It should be understood that there may be waters and variations.

110: biological safety cabinet 111: support
112: plate 113: front cover
114: side cover 120: wave bioreactor
121: seat 122: stirrer
130: gas mixer controller 140: measurement unit
145: bioreactor controller 150: air circulation unit
151: inlet duct 152: intake fan
153: filter 154: circulation duct
161: rotating shaft 162: turbine impeller
163: top magnetic drive 10: disposable bag
20: disposable vessel

Claims (6)

A biosafety cabinet having a support formed at a certain height from the ground and a plate disposed on top of the support, and configured to open and close a part of the front or part of the side to intake outside air and provide an internal space in an aseptic environment;
a wave bioreactor having a seat formed on the top of the plate on which the disposable bag is seated, and a stirrer configured to agitate the cell culture medium of the disposable bag by vibrating the seat up and down or left and right;
a gas mixer controller formed on one side of the biosafety cabinet to control gas supply to the disposable bag;
a measurement unit formed on the seat and connected to each sensor port of the disposable bag to measure pH, DO, temperature, and foam, respectively; and
It is formed on the other side of the biosafety cabinet to control the wave bioreactor, calibrate each sensor of the measurement unit, program the culture environment of the cell culture medium, record and maintain the fermentation process of the cell culture medium, and A bioreactor controller that controls the agitation speed and enables control of DO;
An inlet duct that guides outside air taken in through a partially opened gap of the biosafety cabinet to the upper end of the biosafety cabinet through the back side, an intake fan that takes in outdoor air through the inlet duct, and air passing through the intake fan. Further comprising an air circulation unit composed of a filter for filtering and a circulation duct for circulating the air passing through the filter in the inner space of the biological safety cabinet,
The agitator is composed of a plate coupled to the bottom surface of the seating table, a cam link that swings the plate back and forth or left and right, a driving link interlocked with the cam link, and a motor rotating the driving link, It is configured to rotate forward and reverse at regular intervals on a horizontal plane to stir the cell culture medium of the disposable bag by rotation,
Prior to the seating of the disposable bag on the plate, the internal space of the biosafety cabinet is sterilized with high temperature and high pressure of an autoclave or gamma rays of a gamma ray irradiator.
Disposable cell culture system.
delete According to claim 1,
Characterized in that batch, fed-batch and continuous fermentation are performed through the bioreactor controller,
Disposable cell culture system.
According to claim 1,
Characterized in that the substitution of carbon dioxide and nitrogen is possible through the gas mixer controller,
Disposable cell culture system.
According to claim 1,
Replacing the wave bioreactor, a disposable vessel is seated on the plate, and the gas mixer controller controls gas supply to the disposable vessel,
Disposable cell culture system.
◈Claim 6 was abandoned when the registration fee was paid.◈ According to claim 5,
The disposable vessel includes a rotating shaft formed on a central axis, a turbine impeller formed radially symmetrical at the lower end of the rotating shaft, and a top magnetic drive formed at the lower end of the turbine impeller,
Characterized in that a lower magnetic drive opposite to the upper magnetic drive and a servo motor for rotationally driving the lower magnetic drive are formed at the lower end of the plate.
Disposable cell culture system.

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