KR20120122889A - Isolator and method of migrating culture - Google Patents

Abstract

PURPOSE: An isolator and a transfer method of culture materials are provided to control the property change of culture materials. CONSTITUTION: An isolator (10) comprises a processing chamber, a storage chamber(23), a first opening portion(60), a second opening portion(61), a first door, and a temperature controller(73). When an incubator(15) is installed in the isolator, the first opening portion penetrates a culture room of the incubator and the working area. The first door blocks the second opening portion in order to shut a receiving chamber. The culture material taken out from the incubator is stored in the receiving chamber. When the receiving chamber is shut tightly, and the incubator is removed, the temperature controller controls the temperature of the culture material. [Reference numerals] (100) Container; (101) Container; (20) Decontamiantion gas generating device; (21) Supply device; (22) Progressing chamber; (23) Pass box; (24) Emitting device; (25) Manipulation unit; (26) Control device; (40) Electromagnetic valve; (41) Electromagnetic valve; (42) Pan; (43) Pan; (50) CO2 gas generator; (73) Temperature controller; (74) Humidifier; (80) Inactive device; (81) Inactive device; (A1) Culturing room; (AA) Isolator; (BB,CC) Air

Description

Isolator, culture transfer method {ISOLATOR AND METHOD OF MIGRATING CULTURE}

The present invention relates to an isolator, a method of transferring a culture.

During a series of cell culture steps, a part of all or part of the cultured cells may be transferred to a new container and planted. This operation is called passage, for example, is performed in an isolator having a work chamber in communication with the culture chamber of the culture apparatus when the culture apparatus is installed (see Patent Document 1, for example).

Japanese Patent Laid-Open No. 2011-19762

For example, when culturing the cells after passage again, when the number of containers in which the cells to be cultured are stored is large, there may be a case where the existing culture apparatus cannot store all the containers. In this case, it is necessary to remove the existing culture apparatus from the isolator and install a new culture apparatus with sufficient storage space in the isolator. However, it may take time until the cells are stored in the newly installed culture apparatus after removing the previously installed culture apparatus, so that a property change that is not originally expected in the cells (culture) left in the isolator may occur. There was a case.

This invention is made | formed in view of the said subject, and an object of this invention is to provide the isolator which can suppress the change of the culture property.

In order to achieve the above object, the isolator detachable to the culture apparatus according to an aspect of the present invention includes a working chamber, a storage chamber, and a first opening communicating between the culture chamber of the culture apparatus and the working chamber when the culture apparatus is mounted. A second opening communicating between the work chamber and the storage chamber, a first door closing the second opening to seal the storage chamber, and a culture taken out of the culture chamber are stored in the storage chamber, It is equipped with the temperature control apparatus which adjusts the temperature of the said culture thing accommodated in the said storage chamber, when a storage chamber is sealed and the said culture apparatus is removed.

It is possible to provide an isolator capable of suppressing changes in the properties of the culture.

BRIEF DESCRIPTION OF THE DRAWINGS The figure which shows the structure of the isolator 10 which is 1st Embodiment of this invention.
FIG. 2 is a flowchart showing an example of a process performed by an operator when storing a culture to be subcultured in a new culture device 16. FIG.
3 is a diagram showing the states of the isolator 10 and the culture chamber A1 when passage is performed.
4 is a diagram illustrating a state of the isolator 10 and the culture chamber A1 when the passage is completed.
5 is a diagram illustrating a state of the isolator 10 when the culture chamber A1 is removed.
FIG. 6 is a diagram showing the states of the isolator 10 and the culture chamber B1 when the work chamber 22 and the culture chamber B1 are decontaminated.
7 is a view showing a state of the isolator 10 when the containers 110 and 111 are transferred to the culture chamber B1.
Fig. 8 is a diagram showing the main configuration of the isolator 11 which is the second embodiment of the present invention.
Fig. 9 is a diagram showing the main configuration of the isolator 12 which is the third embodiment of the present invention.

At least the following matters will become apparent from the description of this specification and the accompanying drawings. In addition, sterilization means killing microorganisms and making them as close to sterile as possible, but in the present specification, so-called decontamination, sterilization, sterilization and the like are also included. In addition, a sterile environment is an almost aseptic environment, and decontamination treatment refers to a treatment for realizing the sterile environment, and a substance used for decontamination treatment is called a decontamination substance.

(1st embodiment)

FIG. 1: is a figure which shows the structure of the isolator 10 which is 1st Embodiment of this invention. The isolator 10 is a device for the operator to work with cells and the like in a sterile environment. The culture device 15 is a device for culturing a culture such as cells, and includes a culture chamber A1 for storing the containers 100 and 101 in which the culture is stored, and a door A2 for sealing the culture chamber A1. Equipped.

In addition, the isolator 10 is equipped with a culture device 15, and the isolator 10 includes the decontamination gas generating device 20, the supply device 21, the work chamber 22, the pass box 23, and the discharge device ( 24), the operation part 25 and the control apparatus 26 are comprised.

The decontamination gas generator 20 supplies the hydrogen peroxide gas which is a decontamination gas to the supply apparatus 21.

The supply device 21 is a device for supplying the hydrogen peroxide gas or air outside the isolator 10 to the work room 22 and the pass box 23, and the electromagnetic valves 40 and 41 and the fans 42 and 43. It is configured to include.

Each of the electromagnetic valves 40 and 41 switches the hydrogen peroxide gas or external air based on the control of the control device 26 to supply the fans 42 and 43. The fan 42 supplies hydrogen peroxide gas or external air supplied via the electromagnetic valve 40 to the workroom 22, and the fan 43 passes hydrogen peroxide gas or air supplied from the electromagnetic valve 41. Supply to (23). In addition, the decontamination gas generator 20 and the supply apparatus 21 correspond to a decontamination substance supply apparatus.

The working room 22 is a space for working with a biological material such as a cell. The working chamber 22 is provided with the air filters 50 and 51, the doors 52-54, the working glove 55, and the connection member 56. As shown in FIG. Moreover, the opening part 60 is formed in the wall surface of the side where the cultivation apparatus 15 of the work chamber 22 is installed, and the opening part 61 is formed in the wall surface of the side where the pass box 23 of the work chamber 22 is installed, have.

The air filter 50 is a filter for removing the hydrogen peroxide gas supplied from the fan 42 or the dust contained in the air. The air filter 51 is a filter for removing dust and the like contained in the gas discharged from the working chamber 22. In addition, for example, a HEPA (High Efficiency Particulate Air) filter is used for the air filters 50 and 51.

The door 52 is provided so that opening or closing of the opening part 60 is possible, and the door 53 is provided so that opening or closing of the opening part 61 is possible. Moreover, the door 54 is provided in the front surface of the work room 10 so that opening and closing is possible.

The work glove 55 is provided in an opening (not shown) formed in the door 54 so that an operator can work with cells in the work chamber 22 while the door 54 is closed. In addition, when the doors 52 to 54 are closed, the work room 22 is sealed.

The connecting member 56 is a member for connecting the isolator 10 and the culture apparatus 15, for example, and is provided on the wall surface of the outside of the work chamber 10 so as to surround the opening 60. In addition, if the door 52 (second door) and the door A2 are opened when the culture apparatus 15 is installed in the connecting member 56, the opening 60 is between the work chamber 22 and the culture chamber A1. It communicates through (the 1st opening part). In addition, the work chamber 22 and the pass box 23 communicate with each other through the opening 61 (second opening) when the door 53 (first door) is opened.

The pass box 23 is a space for sterilizing an article such as a culture apparatus or a medium to be carried in the work chamber 22. In addition, when the culture apparatus 15 is removed, for example, in the pass box 23 (storage chamber), a container in which the culture is stored is temporarily stored.

The pass box 23 is provided with the air filters 70 and 71, the door 72, the temperature regulating device 73, the humidifier 74, and the carbon dioxide gas generator 75. As shown in FIG.

The air filter 70 removes the hydrogen peroxide gas supplied from the fan 43 or the dust contained in the air, similar to the air filter 50, and the air filter 71 is similar to the air filter 51. Remove dust, etc. contained in the gas discharged from).

The door 72 (third door) is a door for carrying in articles such as a culture apparatus or a medium into the work room 22. In addition, although the state in which the door 72 was closed is shown in FIG. 1, the articles, such as a culture apparatus, are opening 65 (third opening part) which communicates with the pass box 23 and the exterior when the door 72 is opened. Imported through). In addition, the pass box 23 is sealed in the state in which the doors 53 and 72 are closed.

The temperature adjusting device 73, the humidifier 74 and the carbon dioxide gas generator (CO ₂ gas generator) 75 are cultured to prevent the change of the properties of the culture when the culture is stored in the pass box 23. The temperature of the container in which the water is stored or the environment in the pass box 23 is adjusted.

The temperature adjusting device 73 is, for example, a plate-type thermostat using a Peltier element as a heat source into which a container in which a culture is stored can be loaded. The temperature regulating device 73 is installed at the bottom of the pass box 23, for example, and based on the control of the control device 26, the temperature (temperature of the culture) of the container in which the culture is stored is a predetermined temperature (eg For example, it adjusts to 37 degreeC or 4 degreeC).

The humidifier 74 (humidity adjustment device) adjusts the humidity in the sealed pass box 23 to a predetermined humidity (for example, 90%) based on the control of the control device 26.

The carbon dioxide gas generator 75 (concentration adjusting device) generates the CO ₂ gas based on the control of the control device 26, and sets the CO ₂ concentration in the sealed pass box 23 to a predetermined concentration (eg, 5%).

For this reason, the pass box 23 of this embodiment implements the function of the sterilization chamber in a general isolator, and the function of the storage chamber which can accommodate a culture, preventing the change of the property of a culture.

The discharge device 24 is a device for discharging gas such as hydrogen peroxide gas or air from the work chamber 22 or the pass box 23, and includes the inactivation devices 80 and 81.

Each of the deactivation apparatuses 80 and 81 is provided with a catalyst, detoxifies a gas, and outputs it to the exterior of the isolator 10. FIG.

The operation unit 25 is an operation panel or the like for the operator to set the operation of the isolator 10. In addition, the operation result of the operation part 25 is transmitted to the control apparatus 26. The control device 26 collectively controls the isolator 10 based on the operation result of the operation unit 25.

(Process performed by the operator when transferring the culture to a new culture device)

Here, referring to FIGS. 2 to 7, when there is not enough storage space of the existing culture device 15 (first culture device), the passaged culture is transferred to a new culture device 16 (second). The processing performed by the operator when storing in the cultivation apparatus) will be described. In addition, in the state in which the cultivation apparatus 15 is attached to the isolator 10, it is assumed that all of the culture chamber A1, the work chamber 22, and the pass box 23 are in a sterile environment. The state of each door of (15) shall be a state shown, for example in FIG. In addition, as shown in FIG. 3, the container 110, 111 used at the time of passage is supposed to be carried in the working room 22 beforehand. 3 to 6 illustrate only the main configuration of the isolator 10 for convenience.

First, as shown in FIG. 3, the operator opens the door A2, takes out the container 100 in which the culture is stored, from the culture chamber A1 to the work chamber 22, and performs passage (S100). Specifically, a part of the culture (cells) of the container 100 is transferred to the containers 110 and 111.

When the passage is completed, as shown in FIG. 4, the operator returns the container 100 to the culture chamber A1 and closes the door A2 and the door 52 (S101). In addition, the operator moves the containers 110 and 111 to the pass box 23, and then closes the door 53 to seal the pass box 23 (S102: first and second steps).

After the pass box 23 is sealed, the operator operates the operation unit 25 to prevent the change of the properties of the culture, the temperature of the containers 110 and 111 accommodated in the pass box 23, the pass box 23. The adjustment of the humidity inside and the CO ₂ concentration is started (S103: third step).

And the operator removes the culture apparatus 15 previously installed as shown in FIG. 5 (S104: 4th process), and attaches the new culture apparatus 16 to the isolator 10 (S105: 5th process). In addition, the culture apparatus 16 is provided with the culture chamber B1 and the door B2 which seals the culture chamber B1 similarly to the culture apparatus 15. In this case, it is assumed that no culture is stored in the culture chamber B1, and no decontamination treatment is performed in the culture chamber B1. In the meantime, the temperature in the pass box 23 is continuously adjusted. Therefore, even if the process S104, 105 takes time, for example, the change of the property of the culture accommodated in the container 110, 111 can be prevented.

When the culture apparatus 15 is removed, the operator performs a decontamination process for sterilizing the inside of the culture chamber B1 with the contaminated connection member 56 and the like. Specifically, as shown in FIG. 6, the operator opens the door B2 and the door 52, and then operates the operation unit 25 to decontaminate the decontamination gas generator 20 or the like (S106: 6th process). As a result, the hydrogen peroxide gas generated in the decontamination gas generator 20 is supplied to the work chamber 22 through the air filter 50. Since the hydrogen peroxide gas is also supplied to the culture chamber B1 through the work chamber 22, the work chamber 22 and the culture chamber B1 are sterilized. In addition, since the pass box 23 is sealed during the decontamination treatment, the culture contained in the pass box 23 is not affected by hydrogen peroxide gas.

In addition, after the work chamber 22 and the culture chamber B1 are sterilized, the operator operates the operation unit 25 to complete the adjustment of temperature, humidity, and CO ₂ concentration (S107). As shown in FIG. 7, the operator opens the door 53 and moves the containers 110 and 111 stored in the pass box 23 to the culture chamber B1 (S108: 7th process). As a result, the cultures accommodated in the new containers 110 and 111 can be cultured in the culture chamber B1.

(Second Embodiment)

FIG. 8: is a figure which shows the main structure of the isolator 11 which is 2nd Embodiment of this invention. The isolator 11 is provided with the decontamination gas generator 20 etc. similarly to the isolator 10 shown in FIG. 1, but is abbreviate | omitted here. In addition, the same code | symbol is attached | subjected to the same block in FIG.

The isolator 11 is provided with the storage chamber 200 which is not provided in the isolator 10 shown in FIG. In the work room 210, an opening 300 (second opening) communicating between the storage room 200 and the work room 210, and a door 301 that seals the storage room 200 by closing the opening 300. (First door) is provided. In addition, the temperature adjusting device 73, the humidifier 74, and the CO ₂ gas generator 75 are provided in the storage chamber 200. Thus, for example, when the culture apparatus 15 is removed, the worker can store the container in which the culture is stored in the storage chamber 200. And by changing the environment (temperature, humidity, etc.) of the storage chamber 200, the change of the property of the culture accommodated in the storage chamber 200 can be suppressed.

(Third embodiment)

FIG. 9: is a figure which shows the main structure of the isolator 12 which is 3rd Embodiment of this invention. Like the isolator 10 shown in FIG. 1, the isolator 12 is equipped with the decontamination gas generating apparatus 20 etc., but is abbreviate | omitted here. In addition, the same code | symbol is attached | subjected to the same block in FIG.

When the isolator 12 and the isolator 10 are compared, the isolator 12 is provided with a connecting member 410 having a port 400 to which hydrogen peroxide gas is supplied instead of the connecting member 56. In addition, such a connection member 410 is disclosed by patent document 1 (Unexamined-Japanese-Patent No. 2011-19762, for example).

Then, the temperature regulation device 73, the humidifier 74 and the CO ₂ gas generating device 75 is provided at the studio (22). Thus, for example, when the culture apparatus 15 is removed, the worker can store the container in which the culture is stored in the workroom 22 with the doors 52 and 53 closed. And by changing the environment (temperature, humidity, etc.) of the work chamber 22, the change of the property of the culture accommodated in the work chamber 22 can be suppressed.

In the above, the isolator 10-12 of this embodiment was demonstrated. In the isolator 10, when the culture device 15 is removed, the culture is stored in a pass box 23 provided with the temperature adjusting device 73. Therefore, even when the culture is temporarily stored in the isolator 10, it is possible to prevent a change in the properties of the culture.

In addition, the isolator 10 is provided with a door 52 that closes the opening 60 when the incubator 15 is removed. For this reason, when the culture apparatus 15 is removed, it can suppress that the working chamber 22 is contaminated and that the substance in the working chamber 22 leaks to the outside.

In addition, in the isolator 10, the pass box 23 for carrying an article from the outside is used as a storage chamber in which the culture is temporarily stored. Therefore, space saving can be achieved.

In addition, the pass box 23 of the isolator 10 is provided with a carbon dioxide gas generator 75. Therefore, the change in the properties of the culture can be prevented more than when only the temperature of the culture is adjusted.

In addition, the humidifier 74 is provided in the pass box 23 of the isolator 10. Therefore, the change in the properties of the culture can be prevented more than when only the temperature of the culture is adjusted.

For example, when only the connecting member 410 can be decontaminated, such as the isolator 12, the culture may be temporarily stored in the work room 22 in which the temperature adjusting device 73 is installed. Even in this case, changes in the properties of the culture can be prevented.

In addition, in the isolator 10, when installing a new culture apparatus 16, the culture chamber B1 and the work chamber 22 of the culture apparatus 16 can be sterilized together. Therefore, when installing the new culture apparatus 16, the effort which decontaminates the culture chamber B1 can be reduced previously.

In addition, the said Example is for making an understanding of this invention easy, and does not limit and interpret this invention. The present invention can be changed and improved without departing from the spirit thereof, and the equivalents thereof are included in the present invention.

For example, the temperature adjusting device 73 is a plate-type thermostat using a Peltier element as a heat source capable of directly adjusting the temperature of the container in which the culture is stored, but is not limited thereto. The temperature adjusting device 73 may be, for example, a device for adjusting the temperature in the pass box 23 in the isolator 10. When the temperature in the pass box 23 is adjusted to 37 ° C., for example, the temperature of the culture contained in the pass box 23 is also approximately the same. Therefore, even in this case, it is possible to suppress the change in the properties of the culture. In addition, since the temperature adjusting device 73 should just be able to adjust the temperature in the pass box 23, the heater etc. which are provided in the wall surface of the outer side of the pass box 23 may be sufficient. That is, the temperature adjusting device 73 does not need to be provided inside the pass box 23.

When a part of the working room 22 has the space partitioned so that it may be sealed by a door, if a temperature etc. of the partitioned space can be adjusted, a culture can be stored, suppressing the change of the property of a culture. In addition, the partitioned space where temperature etc. can be adjusted may be provided in the bottom bottom of the work room 22.

10, 11, 12: Isolator
15, 16: culture device
20: decontamination gas generator
21: supply unit
22, 210: Workroom
23: Pass Box
24: discharge device
25: control panel
26: control unit
40, 41: electromagnetic valve
42, 43: Fan
50, 51, 70, 71: air filter
52 to 54, 72, 301, A2, B2: door
55: globe
56, 410: connecting member
60, 61, 65, 300: opening
73: temperature adjusting device
74: humidifier
75: carbon dioxide (CO ₂ ) gas generator
80, 81: deactivator
200: storage room
400: port
A1, B1: culture chamber

Claims (7)

The incubator is a removable isolator,
Workshop,
With a storage room,
A first opening communicating with the culture chamber of the culture apparatus and the working chamber when the culture apparatus is mounted;
A second opening communicating between the work chamber and the storage chamber,
A first door closing the second opening to seal the storage compartment;
When a culture taken out from the culture chamber is stored in the storage chamber, and the storage chamber is sealed, and the culture apparatus is removed, there is provided a temperature adjusting device for adjusting the temperature of the culture stored in the storage chamber. Isolator characterized in that. The isolator according to claim 1, further comprising a second door that closes the first opening when the culture apparatus is removed. According to claim 2, A third opening for communicating between the outside and the storage chamber,
And a third door that opens or closes the third opening,
The third door,
And the second opening is closed by the first door, and is opened when an article is brought into the storage compartment. The carbon dioxide of the storage chamber according to any one of claims 1 to 3, wherein the culture taken out from the culture chamber is stored in the storage chamber, the storage chamber is sealed, and the culture apparatus is removed. An isolator, characterized by further comprising a concentration adjusting device for adjusting the concentration of. The humidity of the storage chamber according to any one of claims 1 to 4, wherein the culture taken out from the culture chamber is stored in the storage chamber, the storage chamber is sealed, and the culture apparatus is removed. Isolator characterized in that it further comprises a humidity control device for adjusting the. The incubator is a removable isolator,
Workshop,
An opening communicating between the culture chamber of the culture apparatus and the work chamber when the culture apparatus is mounted;
A door that closes the opening to seal the work room;
And a temperature adjusting device for adjusting the temperature of the culture stored in the work room when the culture taken out from the culture room is stored in the work room, the work room is closed, and the culture device is removed. Isolator. An isolator in which a first culture device or a second culture device can be mounted, the method for moving a culture containing the culture contained in the first culture chamber of the first culture device to the second culture chamber of the second culture device,
The isolator,
Workshop,
With a storage room,
A first opening communicating between the first or second culture chamber and the working chamber,
A second opening communicating between the work chamber and the storage chamber,
A door that closes the second opening to seal the storage compartment;
Decontamination material supply device for supplying decontamination material to the work room,
And a temperature adjusting device provided in the storage chamber,
A first step of moving the culture of the first culture chamber to the storage chamber when the first culture apparatus is mounted;
A second step of closing the door to seal the storage chamber;
A third step of adjusting the temperature of the culture to the temperature adjusting device;
A fourth step of removing the first culture device,
A fifth step of mounting the second culture apparatus,
A sixth step of decontaminating the work chamber and the second culture chamber in the decontamination material supply device;
And a seventh step of opening the door to move the culture stored in the storage chamber to the second culture chamber.

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