JP2010233482A - Culturing container, and culturing apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a culturing container and culturing apparatus by which the problem of a required preserving place is solved, and also cell-culturing work is efficiently performed by users. <P>SOLUTION: This culturing container 100 including an access port 103 for putting in and taking out a suspension, closing an inside space 102 communicating with the access port 103 from the outside by closing the access port 103, and arranging a plurality of culturing chambers 101 in the inside space 102 at least in one direction, includes divided parts 104 in the inside space 102, performing the closure and release of the closure between the adjacent culturing chambers among the plurality of the culturing chambers 101, and also functioning as the bottom surface part for receiving the suspension. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a culture vessel and a culture apparatus, and is particularly suitable for use when culturing using a plurality of culture vessels.

  In the biotechnology field, cells are cultured. Cell culturing means increasing the number of cells in a predetermined medium, growing them, or changing their properties. Therefore, when culturing cells accompanied by an increase in the number of cells, a large culture area must be secured from the beginning of the culture.

  In order to secure a large culture area, it is sufficient to use a container such as a petri dish or flask having a large culture area (hereinafter collectively referred to as a culture container), but a culture container having a large culture area is used from the beginning of the culture. Is bulky, bothersome and inefficient. Therefore, when the cultured cells increase in the whole culture vessel, a plurality of different culture vessels are used, and further culturing is continued by subcultured the cultured cells to these different culture vessels. Yes.

  However, the use of a plurality of culture vessels requires a storage place for them, and their management becomes complicated.

  The point requiring a storage place can be solved by stacking the plurality of culture vessels in the vertical direction. Non-Patent Document 1 describes a culture vessel (hereinafter referred to as a conventional vessel) having a plurality of chambers in the vertical direction. The conventional container has the same effect as stacking a plurality of culture containers by incorporating a plurality of chambers in the vertical direction.

  The conventional container has an inlet / outlet for allowing cells, culture medium, and the like to enter / exit, and the plurality of chambers communicate with each other through a communication portion that is a space communicating with the inlet / outlet inside the conventional container. . That is, the space inside the conventional container is partly divided into a plurality of spaces by incorporating a plurality of chambers, but the chambers are closed by incorporating the communication portion. It is a connected space.

  FIG. 13 (a) shows a state in which a suspension of cells and medium (symbol 2000) is injected into the conventional container 1000, and FIG. 13 (b) shows that the cells are substantially uniform for each chamber 1001. The state in which the cells are dispensed and the medium is also approximately equally differentiated is shown in FIG. 5C, where the chambers are stacked in the vertical direction and the cells are cultured.

  The transition of the state between the drawings (a) → (b) → (c) of the conventional container 1000 in FIG.

  In FIG. 6A, the suspension 2000 (the wavy line region in the figure) is in a state of being injected into the conventional container 1000 through the inlet / outlet 1003, and the cells and the culture medium are accumulated at the bottom of the conventional container 1000. The conventional container 1000 is placed such that the communication portion 1002 is located below and the chamber 1001 is located above. The chamber 1001 is a plurality of partial spaces formed by the dividing portions 1004 and is a small room that is not closed.

  In FIG. 6B, the entire conventional container 1000 is rotated 180 ° so that the bottom portion and the ceiling portion of the conventional container 1000 in FIG. That is, the communication part 1002 (the hatched area in the figure) is positioned above and the chamber 1001 is positioned below. As a result, the suspension 2000 (the wavy line region in the figure) is divided into each chamber 1001 approximately uniformly by each dividing portion 1004. In addition, in the dispensing of cells, the homogenization of the seeding density is not strict, and it may be uniform to some extent.

  In FIG. 8C, the entire conventional container 1000 is rotated 90 ° so that the surface with the inlet / outlet 1003 is positioned upward. In the figure, each dividing portion 1004 is a plate forming the bottom surface of each chamber. As a result, the suspension 2000 divided substantially uniformly in each chamber spreads to the bottom surface (dividing part 1004) of the chamber in the chamber, and the cells can be cultured. In addition, by providing the rib 1005 in each dividing part 1004, the suspension 2000 can be held in each chamber without flowing out.

"NUNC CATALOGUE 2006-" Naruje Nuku International Co., Ltd. April 2006 production (50-51 pages)

  In the prior art, the point requiring a storage place could be solved by stacking a plurality of culture vessels in the vertical direction.

  However, as described above, the space inside the conventional container 1000 is partially divided into a plurality of spaces, but due to the presence of the communication portion, the chambers are closed spaces. It is a connected space. As a result, for example, when the conventional container 1000 is tilted from the state shown in FIG. 13 (c) toward the state shown in FIG. 13 (a), between the chambers during cell culture (by spilling the suspension). As a result, the amount of suspension and the number of cultured cells differ greatly between the chambers.

  Further, in each chamber, the suspension 2000 can be put only up to the height of the rib 1005. In the state where the suspension 2000 is put to the height of the rib 1005, the suspension 2000 is put in each chamber during cell culture. When the drug is injected, the suspension overflows.

  In the prior art, for example, due to the problems described above, the cell culture work must be performed while always paying attention so that the suspension does not spill from the chamber. Can't be done.

  Therefore, an object of the present invention is to provide a culture container and a culture apparatus that solve the problem of requiring a storage place and allow a user to efficiently perform cell culture work.

  The culture container according to the present invention includes an inlet / outlet for allowing liquid to enter / exit, the internal space leading to the inlet / outlet is closed from the outside by closing the inlet / outlet, and a plurality of culture chambers are provided in the inner space. A culture vessel arranged in at least one direction, wherein a bottom surface portion that closes and releases the closure between adjacent culture chambers of the plurality of culture chambers and receives the liquid in the internal space A dividing part that functions as:

  Since the adjacent culture chambers are mutually closed by the dividing portion, liquid leakage and mixing between the adjacent culture chambers can be prevented, and the user can efficiently perform the cell culture operation. be able to.

  The culture container according to the present invention includes an inlet / outlet for allowing a liquid to enter and exit, and the inner space is closed from the outside by closing the inlet / outlet, and the internal space leading to the inlet / outlet is A dividing part that divides into a plurality of closed culture chambers arranged in at least one direction is provided, and the dividing part has a function as a bottom part for receiving a liquid, and the dividing part divides the internal space. Thus, the plurality of culture chambers closed to each other are formed, and the division part releases the division of the internal space, whereby the closure of the plurality of culture chambers closed to each other is released.

  A plurality of cultures closed to each other, wherein the internal space communicating with the inlet / outlet is divided into a plurality of culture chambers arranged in at least one direction and mutually closed; Since it is divided into chambers, the user can efficiently perform cell culture work.

  The culture container is made of at least a stretchable material, and the dividing portion is formed on a part of the inner surface of the culture container by the expansion and contraction of the material, and the part of the inner surface is in contact with each other. A part divides the internal space to form a plurality of culture chambers that are closed to each other, and the partial contact of the inner surface is released to release the division of the internal space and the parts are closed to each other. The closure of the plurality of culture chambers may be released.

  With this configuration, even when the dividing portion does not divide the internal space, even if the culture vessel does not have a plurality of culture chambers, the dividing portion divides the internal space to form a plurality of culture chambers. Since the plurality of culture chambers are closed to each other, the user can efficiently perform the cell culture work.

  In addition, the first frame body, the second frame body facing the first frame body, and an expansion / contraction portion made of at least a stretchable material, the expansion / contraction portion includes the first frame body and the first frame body. Existing between two frames, joining the first frame and the second frame, and the first frame has walls for forming a plurality of culture chambers closed to each other. Provided, the second frame body is provided with an occlusal portion that meshes with each of the wall portions, and the dividing portion that divides the communication portion that is a part of the internal space includes the wall portion and the occlusion A plurality of culture chambers that are closed to each other by dividing the communicating portion by engaging the wall portion and the occlusal portion corresponding to the wall portion. Then, by releasing the occlusion between the wall portion and the occlusion portion corresponding to the wall portion, the division of the communication portion is released, Closing the plurality of culture chambers that are closed There are or may be released.

  With the above-described configuration having the first frame, the second frame, the expansion / contraction part, the wall part, the occlusion part, and the communication part, even when the dividing part does not divide the internal space (communication part), the plurality of culture chambers Even if a plurality of culture chambers exist, but the culture chambers are not closable, the severing part severes the communication part so that the culturing chambers are closable with each other. Can efficiently perform cell culture work.

  Further, the occlusal part has a communication hole that can be opened and closed, and when the communication part is divided to form a plurality of culture chambers that are closed to each other, by opening the communication hole, adjacent to each other, The closed state may be released in the plurality of culture chambers, and the communication holes may be closed to close each other in the plurality of culture chambers.

With this configuration, in addition to enabling the user to efficiently cultivate cells, it is possible to reduce the number of culture chambers used for cell culture by opening communication holes with the culture container installed vertically. Further, when the plurality of culture chambers arranged in at least one direction by being divided by the dividing portion are stacked in the vertical direction, the plurality of culture chambers are kept closed. May be.

  Thereby, even when it is the state laminated | stacked with respect to the perpendicular direction, a user can perform the culture | cultivation operation | work of a cell efficiently.

  A culture apparatus according to the present invention includes a temperature maintaining apparatus and the culture container according to any one of claims 1 to 4, a pipe connected to the inlet / outlet of the culture container, and the inside of the culture container through the pipe. A liquid injection part for performing liquid injection, a waste liquid part for performing waste liquid from the inside of the culture vessel through the pipe, and an opening / closing part for opening and closing the inlet / outlet, and for dividing the internal space by the dividing part A split drive unit is provided.

  With this configuration, the user can efficiently perform cell culture work. In addition, cells can be cultured with the divided portion having a large area as the bottom surface in a state where cells are approximately uniformly dispensed for each of the plurality of culture chambers and the medium is also approximately equally differentiated. In addition, it is possible to handle the waste liquid in each culture room and collect the detached cultured cells at once, because the user has to deal with each of the culture rooms. To be released. In addition, cell culture can be performed fully or semi-automatically.

  In addition, you may provide the rotation drive part which rotates the said culture container, and the vibration drive part which gives a vibration to the said culture container.

  With this configuration, in addition to enabling the user to efficiently perform the cell culture operation, the cell culture can be performed fully automatically or semi-automatically.

  Closure means that the inside is blocked from the outside, either biologically or chemically, and that heat, moisture, microorganisms, chemicals, gases, etc. do not come and go from the outside to the inside. . Furthermore, a state in which the inside is physically blocked from the outside, that is, a state where force, electromagnetic waves, or the like is not involved with the inside from the outside may be included.

  According to the present invention, it is possible to provide a culture container and a culture apparatus that allow a user to efficiently perform cell culture work.

It is a figure explaining the culture container which concerns on embodiment. It is a figure explaining the culture apparatus which concerns on embodiment. It is a flowchart explaining the culture | cultivation operation | movement which concerns on embodiment. It is a figure explaining the culture container which concerns on embodiment. It is a figure explaining the culture apparatus which concerns on embodiment. It is a flowchart explaining the culture | cultivation operation | movement which concerns on embodiment. It is a figure explaining the culture container which concerns on embodiment. It is a figure explaining the culture apparatus which concerns on embodiment. It is a flowchart explaining the culture | cultivation operation | movement which concerns on embodiment. It is a figure explaining the culture container which concerns on embodiment. It is a figure explaining the culture apparatus which concerns on embodiment. It is a flowchart explaining the culture | cultivation operation | movement which concerns on embodiment. It is a figure explaining the conventional culture container.

Embodiments of the present invention will be described below with reference to the drawings.
[First Embodiment]
(Culture container)
First, FIG. 1 shows a configuration of a culture vessel 100 according to the present embodiment. A suspension is present in the culture vessel 100.

  According to FIG. 1A, the culture vessel 100 includes a main body 106 having an inlet 103a and an outlet 103b (hereinafter, sometimes collectively referred to as the inlet / outlet 103). Therefore, the internal space of the culture vessel 100 (that is, the inside of the main body) communicates with the inlet / outlet 103. By closing the entrance / exit 103, the inside of the main body is closed from the outside. That is, due to this closing property, the inside of the main body 106 is biologically or chemically isolated from the outside world, and heat, moisture, microorganisms, chemical substances, gases, etc. that affect cell culture are outside the world. It is designed not to go to and from the main body. The entire interior of the main body 106 (specifically, the entire interior of the main body 106 excluding the periphery of the entrance / exit 103, etc., but in the following is considered to be the entire interior of the main body 106) is the communicating portion 102. . In the present embodiment, the internal space coincides with the communication portion 102 (therefore, in this embodiment, the inside of the main body 106, that is, the internal space is also referred to as the communication portion 102). The main body 106 may be partially made of a plastic such as glass or polystyrene, or a polymer material, and the culture vessel 100 may include a main body 106 and a gripping part for conveyance.

  Referring to FIGS. 5B to 5C, how to form the dividing portion 104 that divides the communicating portion 102 in this embodiment, and how the dividing portion 104 divides the communicating portion 102 causes the main body. A description will be given of whether the inside of 106 is divided into a plurality of closed spaces (hereinafter referred to as culture chambers) 101 arranged in at least one direction. In the present embodiment and the following embodiments, the description will be made assuming that one direction is the horizontal direction (see (b) to (c) in the figure). Note that the one direction is not limited to the horizontal direction, and may be the vertical direction, and may be regarded as the horizontal direction and the vertical direction even if it is slightly distorted or tilted. Moreover, the circumference direction etc. may be sufficient and it is not restricted to a linear direction. Further, since the culture chambers only need to be arranged in at least one direction, the culture chamber can be formed in an L-shape, T-shape, or W-shape, so that the culture chamber can be arranged in two directions, three directions, or multiple directions It may be arranged.

  The main body 106 is made of a material having at least elasticity, and is made of, for example, a polymer material such as a rubber material or ethylene vinyl acetate. The dividing portion 104 is formed on a part of the inner surface of the main body 106 by expanding and contracting the material, and by contacting a part of the inner surface, the communication portion 102 is divided and the inside of the main body 106 is closed to each other. Divide into a plurality of culture chambers 101.

  According to FIG. 4B, the partition plate B100 is pressed against the main body 106 with respect to the state of FIG. 4A, so that a node 104a is formed on the main body 106 (that is, the partition plate B100 is pressed against the main body 106). A portion formed by expanding and contracting a part of the main body 106 along the partition plate of the partition plate B100 is a node 104a). Further, by pressing the partition plate B100 against the main body 106, as shown in FIG. 3C, the dividing portion 104 is constituted by a part 106a and 106b of the inner surface of the main body 106 (that is, the partition plate B100 is pressed against the main body 106). Thus, the part formed by expanding and contracting a part of the main body 106 along the partition plate of the partition plate B100 is a part 106a of the inner surface of the main body 106), and the parts 106a and 106b of the inner surface are in contact with each other. Then, the communication part 102 is divided and the inside of the main body 106 is divided into a plurality of culture chambers 101 which are closed to each other. Since the entire main body 106 is made of a material having at least elasticity as described above, a part of the main body 106 expands and contracts, a part 106a of the inner surface becomes the dividing portion 104, and a part 106a of the inner surface is the counterpart. By contacting the inner surface part 106 b (the inner surface part 106 b also becomes the dividing portion 104), the communication portion 102 is divided.

  If the main body 106 is partly expanded and contracted so that the inner part 106a becomes the dividing portion 104, the entire main body 106 does not need to be a stretchable material, but only a part of the main body 106. It may be a material having elasticity. Further, the partition plate B100 is pressed against the main body 106 to form the node 104a in the main body 106. However, the main body 106 is twisted while pressing the partition plate B100 against the main body 106, or the main body 106 is used without using the partition plate B100. The node 104b may be formed in the main body 106 by twisting the 106 (FIG. 4D). As a result, the dividing portion 104 is constituted by a part 106c (that is, a node 104b) of the inner surface of the main body 106, and the parts 106c of the inner surface come into contact with each other to divide the communicating portion 102 and And dividing into a plurality of culture chambers 101 which are closed to each other.

  Moreover, when the dividing part 104 releases the division of the communication part 102, the plurality of culture chambers 101 that are mutually closed inside the main body 106 are released from the closed state, and a plurality of culture chambers 101 are formed. Return to space. Specifically, by releasing the contact between the parts 106c on the inner surface, the division of the communication portion 102 is released, and the closure of the plurality of culture chambers 101 that are closed to each other is released.

  When the culture vessel 100 in the state shown in FIG. 5C or FIG. 9D is rotated 90 degrees leftward (or rotated 90 degrees rightward, forward or near to the paper surface, or backward in the paper surface), the dividing unit 104 By being divided, the culture chambers 101 arranged in at least one direction are stacked in the vertical direction (vertical direction). The figure (e) shows the figure which rotated the culture container 100 of the state of the figure (c) 90 degree | times to the left direction. At this time, the plurality of culture chambers 101 are kept closed. Therefore, when the culture vessel 100 is tilted from the stacked state with respect to the vertical direction, the suspension does not spill from each culture chamber 101. The state of being stacked in the vertical direction is not limited to the case where the communication dividing portions 104 between the plurality of culture chambers 101 as shown in the figure are substantially perpendicular to the vertical direction. That is, if it can be considered that the plurality of culture chambers 101 are stacked in the vertical direction, it may be considered that the culture chambers 101 are stacked in the vertical direction.

  In addition, although the six culture chambers 101 are provided in the same figure (b)-(d), the number of culture chambers is not restricted to this.

  As described above, the culture vessel 100 includes the inlet / outlet 103 for allowing the suspension to enter and exit, and the inner space 102 communicating with the inlet / outlet 103 is closed from the outside by closing the inlet / outlet 103, and the inside of the inner space 102. A plurality of culture chambers 101 are arranged in at least one direction, and the interior space 102 is closed between adjacent culture chambers 101 and the closure is released. In addition, it includes a dividing portion 104 that functions as a bottom portion that receives the suspension.

  With this configuration, the plurality of culture chambers 101 can be easily formed in one space inside the main body 106, that is, the communication portions 102 that communicate with the plurality of culture chambers 101 inside the main body 106 are divided, thereby closing each other. The culture chamber 101 can be easily obtained. In addition, it is possible to easily return from the state in which the plurality of culture chambers 101 are formed to the state of one space when it is necessary to insert a reagent, change the medium, or take out the cultured cells. In addition, returning to the state of one space from the state in which the plurality of culture chambers 101 are formed may cancel the division of all the communication portions 102 at one time, or cancel the partial division of the communication portions 102. After that, the remaining communication portion 102 may be disconnected. In this way, it is possible to provide a culture container in which a user can efficiently perform cell culture work.

  In addition, by making the dividing part 104 side with a large area into the bottom face part which receives a suspension, it can culture | cultivate a cell in the state which laminated | stacked each culture chamber, ensuring a culture area. As shown in FIG. 5E, even if the area of the dividing portion 104 that receives the suspension is not large, the vertical direction is applied to the vertical culture vessel 100 to compress the height direction. On the other hand, the culture area can be increased by enlarging the bottom area.

As shown in FIG. 5E, the inner wall of the main body 106 plays the role of this side wall when a side wall is required to hold the suspension without flowing out to the bottom surface portion that receives the suspension. Therefore, it is not necessary to separately provide a rib-like structure that was necessary in the prior art.
(Culture equipment)
Next, FIG. 2 shows a configuration of a culture apparatus 200 using the culture container 100 according to the present embodiment.

  The culture apparatus 200 includes a culture container 100, a pipe P100 connected to the inlet 103a of the culture container 100 when injecting a suspension or the like into the culture container 100, and a valve of the pipe P100 during the injection. An opening / closing part SW100 that opens and closes, and a liquid injection part M100 that injects the liquid into the main body 106 through the pipe P100, and a pipe P101 that is connected to the outlet 103b of the culture container 100 when waste liquid from the culture container 100 is discharged. And an open / close unit SW101 that opens and closes the valve of the pipe P101 in the case of the waste liquid, and a waste liquid part M101 that drains the waste liquid from the inside of the main body 106 through the pipe P101. The inlet 103a and the pipe P100, and the outlet 103b and the pipe P101 are connected by a tube or the like, and the inlet 103a and the pipe P100, and the outlet 103b and the pipe P101 are connected even if the culture vessel is rotated by the rotation drive unit R100 as described later. Is not to leave.

  The liquid injection part M100 is suspended in a culture vessel 100, a medium, physiological saline (hereinafter referred to as PBS), a simple substance of a reagent such as an enzyme, cytokine, protein, or a mixture thereof, or a cell cultured with them. It has a function of injecting carbon dioxide into the infusion solution such as a liquid and the culture vessel 100. The injection is performed using a pump such as a peristaltic pump or a diaphragm pump. Carbon dioxide is normally sealed at an appropriate timing in order to maintain a concentration of 5% for the purpose of maintaining the pH of the medium at an appropriate value. The waste liquid part M101 has a function of draining the waste liquid from the culture vessel 100 to the waste liquid tank. Similarly, the waste liquid is pumped.

  Further, the culture apparatus 200 divides the communicating portion 102 by the dividing portion 104 shown in FIG. 1, so that the partition plate B100 is pressed against the main body 106 as described above (and the main body of the partition plate B100). A separation drive unit B101 for separating from (106).

  Furthermore, the culture apparatus 200 includes a rotation drive unit R100 that rotates the culture vessel, a vibration drive unit V100 that applies vibration to the culture vessel, and a hammer V101. The purpose of rotating the culture vessel is to first divide the suspension into the respective culture chambers by forming a plurality of culture chambers 101 after injecting the suspension with the culture vessel 100 on the side, and thereafter 90 Each of the culture chambers 101 by rotating the culture vessel 100 so that the longitudinal direction of the culture vessel 100 is vertical (that is, the culture chambers 101 arranged in one direction are stacked in the vertical direction). The cells can be cultured with the dividing part 104 having a large area as the bottom surface in a state where the cells are dispensed substantially evenly and the medium is also approximately equally differentiated (in this case, the dividing part 104 is at the bottom). Become). Note that the vertical length may be slightly compressed by applying a force in the vertical direction to the culture vessel 100 standing in the vertical direction. Thereby, in the culture chamber 101, while the height direction is compressed, a bottom area can be expanded and a culture area can be enlarged.

  The purpose of applying vibration to the culture vessel is to peel the cultured cells from the bottom of the culture chamber 101. In addition, when giving a vibration to a culture container instead of giving a vibration to a culture container, you may use a cell peeling agent together.

  In addition, the partition plate B100 is pressed against the main body 106 to form the node 104a in the main body 106, or the main body 106 is twisted to form the node 104b in the main body 106, whereby a plurality of pieces are formed in one space inside the main body 106. The culture chamber 101 can be easily formed. Moreover, it can return to the state of one space easily from the state in which the plurality of culture chambers 101 are formed. That is, the dividing part divides the internal space, thereby forming a plurality of closed spaces inside the main body, and the dividing part cancels the internal space dividing so that the inside of the main body is separated. A plurality of closed spaces can be released. This makes it possible to collect waste liquid from each culture chamber and collect the detached cultured cells. That is, the user is freed from the annoyance and the like that these actions must be performed for each culture room.

  The culture apparatus 200 also includes a temperature maintenance device for maintaining a temperature suitable for cell culture.

  In this way, it is possible to provide a culture device that allows a user to efficiently perform cell culture work. In addition, there is provided a culture apparatus capable of culturing cells with the dividing portion 104 having a large area as a bottom surface in a state where cells are approximately uniformly dispensed for each culture chamber 101 and the medium is also approximately equally differentiated. be able to. In addition, it is possible to handle the waste liquid in each culture room and collect the detached cultured cells at once, because the user has to deal with each of the culture rooms. A released culture device can be provided.

  The control device PC100 controls the operation of the opening / closing unit SW100, the opening / closing unit SW101, the vibration driving unit V100, the dividing driving unit B101, the control of liquid feeding and waste liquid operations, other controls in the culture device 200, and the inside of the culture device 200. The temperature and humidity of each place are managed. The control device PC100 includes a CPU, a ROM storing a culture program, a RAM serving as a work area for each calculation, an OS and various programs, a hard disk storing a culture history, sensor value measurement history, management history, and the like. It consists of storage. The culture program may be stored in the storage. In the control device PC100, an input unit such as a keyboard, a mouse, and a touch panel, which are used to perform various inputs and serve as a culture start button described later, a display on which measurement results, management information, instruction information, warning output, and the like are output, An output unit such as a speaker is connected.

With the configuration described above, it is possible to provide a cell culture apparatus capable of performing cell culture work efficiently and semi-automatically in addition to allowing the user to efficiently perform cell culture work.
(Culture operation)
Next, FIG. 3 shows a flowchart of a culture operation using the culture apparatus 200 according to the present embodiment.

  In step S100, a liquid material such as a culture medium, PBS, or reagent, and a carbon dioxide cylinder are set in the liquid injection part M100 as a pre-operation of the culture operation. Also, a waste liquid tank is set in the waste liquid part M101. A pipe P100 is connected to the inlet 103a, and a pipe P101 is connected to the outlet 103b, and an opening / closing part SW100 opens and closes a valve of the pipe P100.

  In step S <b> 101, the liquid injection part M <b> 100 injects the liquid material and the cell suspension into the culture container 100. At this time, the rotation driving unit R100 may shake the culture vessel 100 so that the suspension is diffused evenly.

  In step S102, the inside of the culture vessel 100 is divided into a plurality of culture chambers 101. That is, the dividing drive unit B101 is used to press against the main body 106 of the partition plate B100, and the communication unit 102 is divided by the dividing unit 104. Thereby, the communication part 102 is divided | segmented and the inside of the main body 106 is divided into the some culture chamber 101 closed mutually.

  In step S103, the rotation drive unit R100 rotates the culture vessel 100 by 90 degrees, and performs rotation driving in which the longitudinal direction of the culture vessel 100 is set to be a vertical direction so that the culture chambers 101 are stacked.

  In step S104, since the plurality of culture chambers 101 are stacked as described above, the cells are dispensed substantially uniformly, and the culture medium is also substantially equally differentiated, so cell culture is started. For example, when a culture start button on the input unit of the control device PC100 is pressed, a culture program stored in the ROM in the control device PC100 is activated, and a heater that keeps the temperature of each culture chamber constant is started. Then, a timer for measuring the culture time is started.

  If it is time to input the reagent or replace the medium by referring to the timer, the process proceeds to the next step S105.

  In step S105, the rotation drive unit R100 rotates the culture vessel 100 by 90 degrees in the direction opposite to that in step S103, so that the plurality of culture chambers 101 coexist sideways. Rotation drive to lie down horizontally.

  In step S106, the inside of the culture vessel 100 is returned from a state in which the plurality of culture chambers 101 are present to a state having one space. That is, the dividing drive unit B101 is used to release the pressing state of the partition plate B100 against the main body 106, and the division of the communication unit 102 by the dividing unit 104 is released. Thereby, the division of the communication unit 102 is released, the plurality of culture chambers 101 closed to each other are integrated, and the inside of the main body 106 is returned to one space.

  In step S107, the opening / closing part SW100 opens and closes the valve of the pipe P100, so that the liquid injection part M100 puts the reagent into the culture vessel 100. At this time, the rotation driving unit R100 may shake the culture vessel 100 so that the reagent is evenly diffused in the suspension.

  In step S107, the opening / closing part SW101 opens and closes the valve of the pipe P101, so that the waste liquid part M101 drains the waste liquid such as the medium from the culture container 100 to the waste liquid tank, and the liquid injection part M100 transfers the suspension to the culture container. The medium may be exchanged by performing the step of reinjecting into 100. Note that the rotation driving unit R100 may shake the culture vessel 100 so that the suspension is diffused evenly.

  In step S108, the interior of the culture vessel 100 is divided into a plurality of culture chambers 101, as in step S102, in order to resume cell culture. That is, the partition plate B100 is pressed against the main body 106 using the dividing drive unit B101, and the communication unit 102 is divided by the dividing unit 104.

  In step S109, as in step S103, the rotation drive unit R100 rotates the culture vessel 100 by 90 degrees to stand the culture vessel 100 in a vertical direction so that the culture chambers 101 are stacked. Rotation drive is performed.

  Thereafter, by referring to the timer, when it is time to add the reagent or replace the culture medium, the steps S105 to S109 are repeated.

  When it is time to end the cell culture by referring to the timer, the operation proceeds to the next cell culture end operation.

  In step S110, the rotation drive unit R100 rotates the culture vessel 100 so that the cells attached to the bottom surface of each culture chamber are separated from the bottom surface.

  In step S111, the vibration drive unit V100 strikes the culture vessel 100 with the hammer V101 so that the cells attached to the bottom surface of each culture chamber are detached from the bottom surface.

  In step S112, the rotation driving unit R100 performs rotation driving to rotate the culture vessel 100 and lay the culture vessel 100 in the horizontal direction so that the plurality of culture chambers 101 coexist sideways.

  In step S113, as in step S106, the inside of the culture vessel 100 is returned from a state in which the plurality of culture chambers 101 are present to a state having one space. That is, the dividing drive unit B101 is used to release the pressing state of the partition plate B100 against the main body 106, and the division of the communication unit 102 by the dividing unit 104 is released.

  In step S114, the opening / closing part SW101 opens and closes the valve of the pipe P101, so that the waste liquid part M101 wastes only the waste liquid such as the culture medium from the culture vessel 100 into the waste liquid tank, and the cells are taken out. Any known method can be used as a method of draining only this waste liquid and taking out the cells. For example, the container may be manually removed and the cells may be removed.

  As described above, when the culture vessel 100 according to the present embodiment is used, a plurality of culture chambers 101 that are closed to each other can be easily formed in a single space inside the main body 106, that is, a plurality of culture chambers 101 that are closed to each other. In addition, it is possible to easily return to the state of one space from the state in which a plurality of culture chambers 101 closed to each other are formed. In other words, when multiple culture chambers closed to each other are necessary for culturing cells, they can be easily formed, while reagents are added and culture media are exchanged, or cultured cells are taken out. When it is desired to eliminate the division of the culture chambers, that is, when it is desired to return to the state of one space from the state in which the plurality of culture chambers 101 are formed, it can be easily restored.

  That is, when the inner surfaces 106a and 106b of the main body 106 come into contact with each other, the communication portion 102 that communicates the plurality of culture chambers 101 inside the main body 106 is divided, and the inside of the main body 106 is divided into a plurality of closed spaces. It is possible to easily form a plurality of culture chambers 101 that are divided, that is, closed in one space inside the main body 106. Moreover, it can return to the state (communication part 102) of one space from the state which formed the several culture chamber 101 closed mutually.

  As described above, since a plurality of culture chambers 101 that are mutually closed can be easily formed in one space inside the main body 106, a culture container and a culture apparatus that allow a user to efficiently perform cell culture work are provided. Can be provided. In addition, since a plurality of culture chambers 101 that are closed to each other can be returned to a single space state, suspensions and reagents can be charged at a time without dispensing each culture chamber. Then, by easily forming a plurality of culture chambers 101 that are closed to each other thereafter, dispensing for each culture chamber can be facilitated. At this time, the suspension and the reagent can be dispensed approximately equally by dividing the communicating portion 102 by the dividing portion 104 described above. Furthermore, since a plurality of culture chambers 101 that are closed to each other can be returned to the state of one space, there is no need to collect each culture chamber at the time of collecting a culture medium or cells, and it can be easily performed at one time. Recovery can be performed.

[Second Embodiment]
In the second embodiment, parts different from the culture container, the culture apparatus, and the culture operation described in the first embodiment will be mainly described. In the present embodiment, the reference numerals and the like in the figure are different from those in the first embodiment, but almost the same part can be easily conceived by replacing it appropriately. In addition, since the thing of the same code | symbol as 1st Embodiment is the same as the culture container, culture | cultivation apparatus, and culture | cultivation operation which were demonstrated in 1st Embodiment, description is abbreviate | omitted.
(Culture container)
First, FIG. 4 shows the configuration of the culture vessel 300 according to this embodiment. A suspension is present in the culture vessel 300.

  According to FIG. 5A, the culture vessel 300 includes a main body 306 having an inlet 303a and an outlet 303b (hereinafter sometimes collectively referred to as an inlet / outlet 303). Therefore, the internal space of the culture vessel 300 (that is, the inside of the main body) communicates with the inlet / outlet 303. By closing the entrance / exit 303, the inside of the main body is closed from the outside. That is, due to this closing property, the inside of the main body 306 is biologically or chemically shielded from the outside world, and heat, moisture, microorganisms, chemical substances, gases, etc. that affect cell culture are outside the world. It is designed not to go to and from the main body. The main body 306 may be made of a plastic material such as glass or polystyrene, or a polymer material, and the culture vessel 300 may include a main body 306 and a gripping part for transport.

  Inside the main body 306, there is a communication portion 302 that is one space communicating with the entrance / exit 303, and the main body 306 includes a first frame body 306a and a second frame body 306b facing the first frame body 306a. And a telescopic portion 306c. The stretchable part 306c exists between the first frame body 306a and the second frame body 306b, and joins the first frame body 306a and the second frame body 306b. The stretchable portion 306c is made of at least a stretchable material, and is made of a polymer material such as a rubber material or ethylene vinyl acetate, for example. The communication portion 302 exists between the first frame 306a and the second frame 306b.

  In the present embodiment and the following embodiments, the communication part is a part of the internal space, and the combination of the communication part and the space in each culture chamber is the internal space. In the present embodiment and the following embodiments, it is a communication part that is a part of the internal space that is divided into the dividing part, but the communication part that is a part of the internal space is divided into the dividing part. This means that the internal space is divided by the dividing portion.

  The first frame body 306a has a wall portion 304a for constituting a plurality of culture chambers that are closed together, and the second frame body 306b has an occlusion portion 304b that engages with each of the wall portions 304a. The dividing part 304 that is provided and divides the communication part 302 is composed of the wall part 304a and the occlusal part 304b. Note that the culture vessel 300 may include a gripping part for conveyance in addition to the main body 306.

  In FIG. 6A, the culture chamber 301 is divided (not closed) and has a communication part 302. In FIG. 5B, the expansion / contraction part 306c expands and contracts to the wall part. When the occlusal portion 304b corresponding to the wall portion 304a is engaged, the dividing portion 304 divides the communicating portion 302, and the inside of the main body 306 is a culture that is a plurality of closed spaces arranged in at least one direction. The room 301 is divided. Each culture chamber 301 has a structure surrounded by a first frame body 306a, a second frame body 306b, a wall portion 304a, and an occlusal portion 304b, and the culture chambers 301 at both ends are further surrounded by an expansion / contraction portion 306c. It has a structured.

  According to FIG. 5B, the expansion and contraction portion 306c contracts and the first frame body 306a approaches the second frame body 306b, so that the wall portion 304a and the occlusion portion 304b are engaged, and the first frame body 306a and the second frame body 306a are in contact with each other. The communicating portion 302 existing between the frames 306b is divided by the dividing portion 304. In the second embodiment, unlike the first embodiment, the dividing portion 304 is present from the beginning, and the main body 306 is divided into a plurality of culture chambers 301. Inside the main body 306, a communication portion 302 and each culture chamber 301 exist.

As described above, the culture vessel 300 includes the inlet / outlet 303 for allowing the suspension to enter and exit, and the inner space (in this embodiment, the communication unit 302 and each culture) that is connected to the inlet / outlet 303 by closing the inlet / outlet 303. A culture vessel 300 that is closed from the outside and has a plurality of culture chambers 301 arranged in at least one direction in the internal space, and a plurality of cultures in the internal space. In addition to closing and releasing the adjacent culture chambers in the chamber 301, a dividing portion 304 that functions as a bottom surface portion that receives the suspension is provided. With this configuration, a plurality of spaces are provided in one space inside the main body 306. The culture chambers 301 that are closed to each other can be easily obtained by forming the culture chambers 301 easily, that is, by eliminating the communication portions 302 that communicate the plurality of culture chambers 301 inside the main body 306 Can. In addition, it is possible to easily return from the state in which the plurality of culture chambers 301 are formed to the state of one space when it is necessary to add a reagent, change a medium, or take out cultured cells. That is, the dividing part divides the internal space, thereby forming a plurality of closed spaces inside the main body, and the dividing part cancels the internal space dividing so that the inside of the main body is separated. The plurality of spaces closed to each other can be released from the closed state, and the plurality of spaces can be returned to the state of one space. In addition, returning to the state of one space from the state in which the plurality of culture chambers 301 are formed may cancel the division of all the communication portions 302 at one time, or release the partial division of the communication portion 302 Then, it may be configured such that the division of the remaining communication portion 302 is released.

  In this way, it is possible to provide a culture container in which a user can efficiently perform cell culture work.

  In addition, the figure which rotated the culture container 300 of the state of the figure (b) 90 degree | times to the left in the figure (c) is shown. As shown in FIG. 5C, the cell is cultured in a state where the culture chambers are stacked while securing the culture area by making the dividing portion 304 side having a large area the bottom surface for receiving the suspension. Can do.

  The culture vessel 300 in the state of FIG. 5C is in a state in which the culture chambers 301 arranged in at least one direction by being divided by the dividing unit 304 are stacked in the vertical direction. At this time, the plurality of culture chambers 301 are kept closed. Therefore, when the culture vessel 300 is tilted from the stacked state with respect to the vertical direction, the suspension does not spill from each culture chamber 301. The state of being stacked in the vertical direction is not limited to the case where the communication dividing portions 304 between the plurality of culture chambers 301 as shown in the figure are substantially perpendicular to the vertical direction. That is, if it can be considered that the plurality of culture chambers 301 are stacked in the vertical direction, it may be considered that the culture chambers 301 are stacked in the vertical direction.

  Note that, as shown in FIG. 6C, a side wall is required to hold the suspension without flowing out to the bottom surface portion that receives the suspension, and the inner wall of the main body 306 plays the role of this side wall. Yes. Therefore, it is not necessary to separately provide a rib-like structure that was necessary in the prior art.

In the figure, six culture chambers 301 are provided, but the number of culture chambers is not limited to this.
(Culture equipment)
Since the culture apparatus in the second embodiment is almost the same as the culture apparatus described in the first embodiment, different parts will be mainly described.

  FIG. 5 shows a configuration of a culture apparatus 400 using the culture vessel 300 according to the present embodiment.

  In order to divide the communicating part 302 by the dividing part 304, the culture device 400 contracts the expansion / contraction part 306c described with reference to FIG. And a wall drive part B200 for separating the wall part 304a from the occlusion part 304b).

  The culture apparatus 400 includes a rotation drive unit R100 that rotates the culture vessel. The purpose of rotating the culture vessel is as follows. Initially, the culture container 300 is laid down with the communicating portion 302 facing down, and the suspension is injected into the communicating portion 302 with the wall portion 304a separated from the occlusal portion 304b, and then the wall portion 304a and the occlusal portion. The culture chamber 301 is formed by occlusion of 304b, so that the suspension is substantially equally differentiated into each culture chamber, and then rotated 90 degrees so that the longitudinal direction of the culture vessel 300 is set to the vertical direction. This is because the cells can be cultured with the wall 304a (dividing part 304) having a large area as the bottom surface in a state in which the cells are dispensed approximately uniformly every 301 and the medium is substantially equally differentiated ( In this case, the dividing portion 304 is the bottom).

  The culture apparatus 400 also includes a temperature maintenance device for maintaining a temperature suitable for cell culture.

From the above, it is possible to provide a culture apparatus that allows the user to efficiently perform cell culture work. In addition, there is provided a culture apparatus capable of culturing cells with the dividing portion 304 having a large area as a bottom surface in a state where cells are approximately uniformly dispensed for each culture chamber 301 and the medium is also substantially equally differentiated. be able to. In addition, it is possible to handle the waste liquid in each culture room and collect the detached cultured cells at once, because the user has to deal with each of the culture rooms. A released culture device can be provided. Furthermore, the above configuration can provide a cell culture apparatus capable of performing cell culture fully or semi-automatically.
(Culture operation)
Since the flow of the culture operation in the second embodiment is almost the same as the flow of the culture operation described in the first embodiment, different portions will be mainly described. That is, in this embodiment, step S102 is replaced with step S202, step S106 is replaced with step S206, step S108 is replaced with step S208, and step S113 is replaced with step S213. The other steps are the contents of the first embodiment as the culture vessel 300, What is necessary is just to interpret suitably according to the culture apparatus 400.

  FIG. 6 shows a flowchart of a culture operation using the culture apparatus 400 according to the present embodiment.

  In step S202, the plurality of culture chambers 101 are closed. That is, when the partitioning drive unit B101 is used to engage the wall portion 304a and the occlusion portion 304b corresponding to the wall portion 304a, the separation portion 304 divides the communication portion 302, and the interior of the main body 306 is closed to each other. The culture room 301 is divided into spaces.

  In step S206, the closed state between the plurality of culture chambers 101 is released. That is, the separation of the communicating portion 302 is released by separating the wall portion 304a from the occlusal portion 304b using the dividing drive unit B101, and the culture chamber 301, which is a plurality of closed spaces, is integrated, Return to one space.

  In step S208, the plurality of culture chambers 101 are closed as in step S202 in order to resume cell culture. That is, when the partitioning drive unit B101 is used to engage the wall portion 304a and the occlusion portion 304b corresponding to the wall portion 304a, the separation portion 304 divides the communication portion 302, and the interior of the main body 306 is closed to each other. The culture room 301 is divided into spaces.

  In step S213, the closed state between the plurality of culture chambers 101 is released as in step S206. That is, the separation of the communicating portion 302 is released by separating the wall portion 304a from the occlusal portion 304b using the dividing drive unit B101, and the culture chamber 301, which is a plurality of closed spaces, is integrated, Return to one space.

  As described above, when the culture vessel 300 according to the present embodiment is used, the wall 304a and the occlusal portion 304b corresponding to the wall 304a are engaged to exist between the first frame 306a and the second frame 306b. The communication part 302 is divided to divide the inside of the main body 306 into a plurality of closed spaces, that is, a plurality of culture chambers 301 closed to each other can be easily formed inside the main body 306, and are closed to each other. In addition, it is possible to easily return to the state where the communication portion 302 exists from the state where the plurality of culture chambers 301 are formed. In other words, when multiple culture chambers closed to each other are necessary for culturing cells, they can be easily formed, while reagents are added and culture media are exchanged, or cultured cells are taken out. When it is desired to eliminate the division of the culture chambers, that is, when it is desired to return from the state in which the plurality of culture chambers 301 are formed to the state in which the communication portion 302 exists, it can be easily returned.

  A plurality of culture chambers 301 that are mutually closed inside the main body 306 can be easily formed, that is, the communication portions 302 that communicate with the plurality of culture chambers 301 inside the main body 306 are divided, so that Obtainable. Moreover, it can return to the state in which the communication part 302 exists from the state which formed the some culture chamber 301 closed mutually.

  As described above, since a plurality of culture chambers 301 that are mutually closed can be easily formed in one space inside the main body 306, a culture container and a culture apparatus that allow a user to efficiently perform cell culture work are provided. Can be provided. In addition, since a plurality of culture chambers 301 closed to each other can be returned to the state of one space, suspensions and reagents can be charged at a time without dispensing each culture chamber. Then, by easily forming a plurality of culture chambers 301 that are closed to each other thereafter, dispensing for each culture chamber can be facilitated. At this time, the suspension and the reagent can be dispensed approximately evenly by dividing the communicating portion 302 by the dividing portion 304 described above. Furthermore, since a plurality of culture chambers 301 closed to each other can be returned to the state of one space, there is no need to collect each culture chamber at the time of collecting the culture medium and cells, and it can be easily performed at one time. Recovery can be performed.

[Third Embodiment]
Since the third embodiment is almost the same as the culture vessel, the culture apparatus, and the culture operation described in the second embodiment, different parts will be mainly described. In the present embodiment, the reference numerals and the like in the drawing are different from those in the second embodiment, but almost the same part can be easily conceived by appropriately replacing it. In addition, since the thing of the same code | symbol as 2nd Embodiment is the same as the culture container, culture | cultivation apparatus, and culture | cultivation operation which were demonstrated in 2nd Embodiment, description is abbreviate | omitted.
(Culture container)
FIG. 7 shows a configuration of a culture vessel 500 according to this embodiment.

  In this embodiment, each occlusal portion 304b is provided with a communication hole (not shown, no symbol), and the wall portion 304a and the occlusal portion 304b, which are the dividing portions 304, are engaged with each other to form the first frame body 306a and the second frame. Even if the communication part 302 existing between the bodies 306b is divided by the dividing part 304, the culture chambers 501 adjacent to each other across the communication hole communicate with each other through the communication hole (hereinafter referred to as communication). Each communication hole is provided with a communication hole sealing valve 504. By closing the communication hole with the communication hole sealing valve 504, communication between adjacent culture chambers 501 is disconnected. That is, by separating the communication in the communication hole by the communication hole sealing valve 504, the culture chambers 501 adjacent to each other across the communication hole are closed. On the contrary, the closed state between the adjacent culture chambers 501 is released by opening the communication hole.

  FIG. 6A shows a state in which all the communication hole sealing valves 504 are closed. When the wall portion 304a and the occlusal portion 304b are engaged in this state, the culture chambers 501 are arranged in at least one direction and are closed from each other. Therefore, when the wall portion 304a and the occlusal portion 304b are engaged and stood as shown in FIG. 5B, a suspension is present in each culture chamber 501.

  FIG. 5C shows a state in which the communication holes at both ends are opened, every other communication hole is opened, and the remaining communication holes are closed. When the wall portion 304a and the occlusal portion 304b are engaged in this state, only the adjacent culture chambers 401 communicate with each other. Other than the adjacent culture chamber 501 is in a closed state. Therefore, when standing as shown in FIG. 4D, the suspension flows from the upper culture chamber to the lower culture chamber through the communication hole, so that there is a suspension in every other culture chamber 501. (In this case, the number of culture chambers in which the suspension is present is ½ of the total number). As will be described later, in an actual culture operation, a predetermined communication hole is opened at the initial stage of culture (see step S302 and the like described later), and as the culture proceeds, the communication hole sealing valve 504 is moved to perform a predetermined operation. The communication hole is closed to expand the culture area (see step S308 and the like described later). Explaining in the figure, the culture is in the state of FIG. (D) at the initial stage of culture (in this figure, the first, third and fifth communication hole sealing valves 504 from the top correspond to the predetermined communication holes. ) As the culture proceeds, the state finally shifts to the state shown in FIG.

  With the above configuration, the number of culture chambers used for cell culture can be reduced by opening the communication holes in a state where the culture container is vertically installed. That is, by opening the communication hole, the culture area used for cell culture can be reduced, and by opening / closing the communication hole by the communication hole sealing valve 504, increase / decrease of the culture area used for cell culture can be adjusted.

  In cell culture, the culture area may depend on the number of cells contained in the suspension, and if the cell area is too large, cell culture may be adversely affected. Therefore, the communication hole sealing valve 504 is initially opened with a communication hole, the culture area is narrowed, and as the culture proceeds, the communication hole is closed with the communication hole sealing valve 504 to expand the culture area. We have to go. The predetermined communication hole sealing valve 504 to be opened can be determined from the necessary culture area. By using the culture container of this embodiment, it is possible to cope with such a culture method.

  As described above, it is possible to provide a culture container in which a user can efficiently perform cell culture work.

In addition, in the same figure (a)-(d), although six culture chambers 501 are provided, the number of culture chambers is not restricted to this.
(Culture equipment)
The culture apparatus in the third embodiment is almost the same as the culture apparatus described in the second embodiment, and therefore, different parts will be mainly described. .

FIG. 8 shows a configuration of a culture apparatus 600 that uses the culture vessel 500 according to the present embodiment. In FIG. 8, the illustration of the rotation drive unit R100, the vibration drive unit V100, and the hammer V101 described in the second embodiment is omitted for convenience of description. It has the same function as the embodiment (hereinafter, refer to the rotation drive unit R100, the vibration drive unit V100, and the hammer V101 in FIG. 5).
The culture apparatus 600 has a valve drive unit B500 that operates the communication hole sealing valve 504 to open and close each communication hole of the culture vessel 500.

  The culture apparatus 600 also includes a temperature maintenance device for maintaining a temperature suitable for cell culture.

As described above, it is possible to provide a culture apparatus that allows a user to efficiently perform cell culture work. In addition, there is provided a culture apparatus capable of culturing cells with the dividing portion 304 having a large area as a bottom surface in a state where cells are approximately uniformly dispensed for each culture chamber 501 and the medium is substantially equally differentiated. be able to. In addition, it is possible to handle the waste liquid in each culture room and collect the detached cultured cells at once, because the user has to deal with each of the culture rooms. A released culture device can be provided. Furthermore, the above configuration can provide a cell culture apparatus capable of performing cell culture fully or semi-automatically.
(Culture operation)
Since the flow of the culture operation in the present embodiment is almost the same as the flow of the culture operation described in the second embodiment, different parts will be mainly described. That is, in the present embodiment, step S202 is replaced with step S302, and step S208 is replaced with step S308, and the contents of the second embodiment may be appropriately interpreted according to the culture vessel 500 and the culture apparatus 600 in other steps.

  FIG. 9 shows a flowchart of a culture operation using the culture apparatus 400 according to the present embodiment.

  In step S302, in addition to the operation of step S202, the valve drive unit B500 moves the communication hole sealing valve 504 to open a predetermined communication hole. For example, as shown in FIG. 7D, the communication hole corresponding to the first, third, and fifth communication hole sealing valves 504 from the top may be a predetermined communication hole.

  In step S308, in addition to the operation of step S208, the valve drive unit B500 moves the communication hole sealing valve 504 to expand the culture area as the culture proceeds. Specifically, when the culture vessel is in the horizontal state, the communication hole is closed, and the rotation drive unit R100 rotates the culture vessel to change the culture vessel from the horizontal state to the vertical state, thereby suspending the suspension in each culture chamber. Is satisfied and the culture area is expanded.

  While repeating steps S105 to S109, as the culture proceeds, the communication hole sealing valve 504 is moved to close the communication hole. The timing for closing the communication hole in the time axis direction is determined from the result of counting the number of cells and the like, and the communication hole is closed at a timing when the number of cells is considered to reach a predetermined upper limit value.

  As described above, the culture container 500 and the culture apparatus 600 of the present embodiment have the culture container 300 and the culture apparatus 400 of the present embodiment, which allow the user to efficiently perform cell culture work. In addition to the effect of providing the device, the connection hole sealing valve 504 opens and closes the connection hole, thereby adjusting the increase / decrease of the culture area used for cell culture.

[Fourth Embodiment]
The culture apparatus in the fourth embodiment has a form in which two culture containers described in the third embodiment are combined. Therefore, in the present embodiment, mainly the portions different from the description in the third embodiment and the portions that require addition in the culture container, the culture apparatus, and the culture operation will be mainly described. In the present embodiment, the reference numerals and the like in the drawings are different from those in the third embodiment, but almost the same part can be easily conceived by appropriately replacing it. In addition, since the thing of the same code | symbol as 3rd Embodiment is the same as the culture container, culture | cultivation apparatus, and culture | cultivation operation which were demonstrated in 3rd Embodiment, description is abbreviate | omitted.
(Culture container)
FIG. 10 shows a configuration of a culture vessel 700 according to this embodiment.

  The culture apparatus 700 in the present embodiment uses two culture containers 500 described in the third embodiment (referred to as culture containers 5001 and 5002), and the first frame 306a (of the culture container 5001) of each of the culture containers 5001 and 5002. The first frame body 306a1 and the culture vessel 5002 are referred to as the first frame body 306a2), and the side is associated with each other.

  In the present embodiment, the second frame 306b, the two expansion / contraction portions 306c, the wall portion 304a, the occlusion portion 304b, the communication portion 302, the inlet / outlet 303, the communication hole sealing valve, which are referred to in the culture vessel 500 described in the third embodiment. 504 is represented as follows.

  The culture container 5001 is represented by a second frame 306b1, two telescopic portions 306c1, a wall 304a1, an occlusion portion 304b1, a communication portion 3021, an inlet / outlet 3031 (inlet 3031a and outlet 3031b), and a communication hole sealing valve 5041. Yes.

  The culture vessel 5002 is represented as a second frame 306b2, two expansion / contraction portions 306c2, a wall portion 304a2, an occlusion portion 304b2, a communication portion 3022, an inlet / outlet 3032 (inlet 3032a and outlet 3032b), and a communication hole sealing valve 5042. Yes.

  A slide valve 706e exists between the two first frame bodies 306a1 and 306a2, and two container culture chambers 701 (the culture container 5001 is a culture chamber 7011 and the culture container 5002 is a culture chamber). 7012), each hole 706d can be opened and closed by sliding the slide valve 706e. By opening the hole 706d, a suspension or the like can pass between the adjacent culture chambers 7011 and 7012 (same meaning as “contact” in the third embodiment), and the hole 706d is closed. Thus, the passage between the culture chamber 7011 and the culture chamber 7012 is closed, and by closing the hole 706d, the culture chamber 7011 and the culture chamber 7012 of the two containers are closed. That is, the closed state between the culture chambers 7011 and 7012 of the two containers is released by opening the hole 706d.

  As described above, the culture apparatus 700 is an integrated culture container including the culture container 5001, the culture container 5002, and the slide valve 706e.

  FIG. 5A is a diagram in which the culture apparatus 700 is in the horizontal direction with all the communication hole sealing valves 5041 and 5042 closing the communication holes and the slide valve 706e closing the hole 706d. In this state, when the wall portion 304a1 and the occlusal portion 304b1, and the wall portion 304a2 and the occlusal portion 304b2 are engaged, all the culture chambers 7011 are arranged in one direction and are in a closed state. All the culture chambers 7012 are also arranged in one direction and are closed with respect to each other. In FIG. 9A, the suspension exists only on the culture container 5002 side. When the wall portion 304a1 and the occlusal portion 304b1 and the wall portion 304a2 and the occlusal portion 304b2 are occluded and are erected as shown in FIG. 5B, the suspension exists only in each culture chamber 7012. Thereafter, when the slide valve 706e is slid to open the hole 706d, the closed state between the adjacent culture chambers 7011 and 7012 is released, so that the suspension exists only in each culture chamber 7012. A suspension is present in all of the culture chambers 7011 and 7012.

  With the above configuration, the number of culture chambers used for cell culture can be reduced by closing the hole 706d by sliding the slide valve 706e in a state where the culture vessel is vertically installed. That is, by closing the hole, the culture area used for cell culture can be reduced, and by opening / closing the hole 706d by the slide valve 706e, the increase / decrease of the culture area used for cell culture can be adjusted.

  Further, the hole 706d is closed by the slide valve 706e, the reagent or the like is poured into the culture chamber 7011, and the suspension is stored in the culture chamber 7012. The hole 706d is opened by the slide valve 706e. Thus, the reagent in the culture chamber 7011 and the suspension in the culture chamber 7012 can be mixed, and then the culture area is expanded using both the culture chamber 7011 and the culture chamber 7012, and the culture is continued. be able to.

  From the above, it is possible to provide a culture container in which a user can efficiently perform cell culture work.

In addition, in the same figure (a)-(b), although the culture chambers 7011 and 7012 are each provided, the number of culture chambers is not restricted to this.
(Culture equipment)
Since the culture apparatus in the fourth embodiment is almost the same as the culture apparatus described in the third embodiment, different parts will be mainly described.

  FIG. 11 shows a configuration of a culture apparatus 800 that uses the culture vessel 700 according to the present embodiment. In FIG. 11, the illustration of the rotation drive unit R100, the vibration drive unit V100, and the hammer V101 described in the second embodiment is omitted for convenience of description, but the culture apparatus of this embodiment also includes the second. It has the same function as the embodiment (hereinafter, refer to the rotation drive unit R100, the vibration drive unit V100, and the hammer V101 in FIG. 5).

  The culture apparatus 800 includes a pipe P800 connected to the inlets 3031a and 3032a of the culture container 700 when injecting a suspension or the like into the culture container 700, and a pipe P800 on the inlet / outlet 3031a side during the injection. An opening / closing part SW7001 for opening / closing the valve and an opening / closing part SW7002 for opening / closing the valve of the pipe P800 on the inlet / outlet 3032a side during the injection are provided. In addition, a pipe P801 connected to the inlet / outlet 3031b and 3032b of the culture container 700 when the waste liquid from the culture container 700 is disposed, and an opening / closing part SW7011 that opens and closes a valve of the pipe P801 on the inlet / outlet 3031b side when the waste liquid is present. In the case of the waste liquid, an opening / closing part SW7012 for opening / closing a valve of the pipe P801 on the inlet / outlet 3032b side is provided. In the present embodiment, as shown in the figure, there are two valves for the pipe P800 and two for the pipe P801.

  Further, in order to divide the communicating portions 3021 and 3022 by the dividing portions 304a1 and 304a2, the culture apparatus 800 contracts the expansion and contraction portions 306c1 and 306c2 to connect the wall portion 304a1 and the occlusal portion 304b1, and the wall portion 304a2 and the occlusal portion 304b2. Dividing drive parts B271 and B272 for engaging (and extending the telescopic parts 306c1 and 306c2 to separate the wall part 304a1 from the articulating part 304b1 and the wall part 304a2 from the articulating part 304b2) are provided.

  In addition, the culture apparatus 800 includes a valve drive unit B571 that operates the communication hole sealing valve 5041 and a valve drive unit B572 that operates the communication hole sealing valve 5042 to open and close each communication hole of the culture vessel 700. In order to open and close each hole 706d of the culture vessel 700, a valve drive unit B573 for sliding the slide valve 706e is provided.

  The culture apparatus 800 also includes a temperature maintenance device for maintaining a temperature suitable for cell culture.

From the above, it is possible to provide a culture apparatus that allows the user to efficiently perform cell culture work. In addition, there is provided a culture apparatus capable of culturing cells with the dividing portion 304 having a large area as a bottom surface in a state where cells are approximately uniformly dispensed for each culture chamber 701 and the medium is also substantially equally differentiated. be able to. In addition, it is possible to handle the waste liquid in each culture room and collect the detached cultured cells at once, because the user has to deal with each of the culture rooms. A released culture device can be provided. Furthermore, the above configuration can provide a cell culture apparatus capable of performing cell culture fully or semi-automatically.
(Culture operation)
Since the flow of the culture operation in the present embodiment is almost the same as the flow of the culture operation described in the third embodiment, different parts will be mainly described. That is, in this embodiment, step S302 is replaced with step S402, and step S308 is replaced with step S408, and the other steps may be appropriately interpreted in accordance with the contents of the third embodiment according to the culture vessel 700 and the culture apparatus 800.

  FIG. 12 shows a flowchart of a culture operation using the culture apparatus 800 according to this embodiment.

  In step S402, in addition to the operation of step S302, the valve drive units B571 and B572 move the communication hole sealing valves 5041 and 5042 to open predetermined communication holes.

  In step S408, in addition to the operation of step S308, the valve drive units B571 and B572 move the communication hole sealing valves 5041 and 5042 to expand the culture area as the culture progresses. Alternatively, the valve drive unit B573 slides the slide valve 706e to mix the reagent in the culture chamber 7011 and the suspension in the culture chamber 7012 and expand the culture area as the culture proceeds. Specifically, when the culture vessel is in the horizontal state, the communication hole is closed, and the rotation drive unit R100 rotates the culture vessel to change the culture vessel from the horizontal state to the vertical state, thereby suspending the suspension in each culture chamber. Is satisfied and the culture area is expanded. Further, by opening the hole 706d when the culture container is in the vertical state, for example, the reagent in the culture chamber 7011 and the suspension in the culture chamber 7012 are mixed, and the culture area is expanded.

  While repeating steps S105 to S109, as the culture proceeds, the communication hole sealing valve 504 is moved to close the communication hole. Further, the slide valve 706e is slid to open the hole 706d. How to close the communication hole in the time axis direction and how to open the hole 706d are determined from the results of experiments and the like.

  As described above, the culture container 700 and the culture apparatus 800 of the present embodiment have the culture container 500 and the culture apparatus 600 of the present embodiment, which allow the user to efficiently perform cell culture work. In addition to the effect of providing the device, the connection hole sealing valve 504 opens and closes the connection hole, thereby adjusting the increase / decrease of the culture area used for cell culture.

  In addition, since the reagent can be mixed only in the culture container, the container is more closed and the risk of contamination in the culture work can be reduced.

In the present embodiment, two culture containers are combined (culture container 5001 and culture container 5002). However, three culture containers or three or more culture containers may be combined. .
[Other Embodiments]
Control of the culture apparatuses 200, 400, 600, and 800 in the present embodiment can be realized in hardware by a CPU, memory, or other LSI of any computer. In terms of software, it is realized by a program having a cell culture function loaded in the RAM. For example, the control device PC100 in FIGS. 2, 5, 8, and 11 shows a functional configuration such as a computer, and is realized by this configuration. However, it goes without saying that these functional blocks can be realized in various forms such as hardware only, software only, or a combination thereof.

  That is, a cell culture program for operating a general-purpose computer used in the culture devices 200, 400, 600, and 800 as the control device PC100 is shown in the flow chart of the cell culture operation shown in FIGS. Used as appropriate.

  The embodiments of the present invention can be appropriately modified in various ways within the scope of the technical idea shown in the claims. The above embodiment is merely one embodiment of the present invention, and the meaning of the term of the present invention or each constituent element is not limited to that described in the following embodiment.

100, 200, 300, 400 Culture vessel 101, 201, 301, 401 Culture chamber 102, 202, 302, 402 Communication part (internal space)
103, 203, 303, 403 Inlet / outlet 104, 204, 304, 404 Dividing part, bottom part 106, 206, 306, 406 Main body 504 Communication hole sealing valve B101, B200, B500, B571, B572 Dividing drive part

Claims (6)

A culture comprising an inlet / outlet for allowing liquid to enter and exit, wherein an internal space leading to the inlet / outlet is closed from the outside by closing the inlet / outlet, and a plurality of culture chambers are arranged in at least one direction in the internal space A container,
A culture vessel provided with a dividing part that functions as a bottom part for receiving the liquid while closing and releasing the closure between adjacent culture rooms among the plurality of culture rooms in the internal space.
The culture vessel is made of at least a stretchable material,
The dividing part is formed on a part of the inner surface of the culture vessel by expanding and contracting the material,
The inner part is divided by the part of the inner surface contacting each other, forming a plurality of culture chambers closed to each other,
The culture container according to claim 1, wherein the inner space is disconnected by releasing the contact of a part of the inner surface, and the closure of the plurality of culture chambers closed to each other is released.
It is composed of a first frame, a second frame facing the first frame, and at least a stretchable part made of a stretchable material,
The expansion / contraction part exists between the first frame and the second frame, and joins the first frame and the second frame,
The first frame body is provided with a wall portion for constituting a plurality of culture chambers closed to each other,
The second frame body is provided with an occlusal portion that engages with each of the wall portions,
The dividing portion that divides the communication portion that is a part of the internal space is composed of the wall portion and the occlusal portion,
The expansion and contraction part expands and contracts the communication part by engaging the wall part and the occlusion part corresponding to the wall part to form the plurality of culture chambers closed to each other,
The occlusion between the wall portion and the occlusal portion corresponding to the wall portion is released to release the division of the communication portion, and the closure of the plurality of culture chambers closed to each other is released. The culture container according to 1.
The occlusal portion has a communication hole that can be opened and closed,
When the communication part is divided and the plurality of culture chambers closed to each other are formed, by opening the communication hole, the closed state is released in the plurality of adjacent culture chambers, The culture container according to claim 3, wherein the communication holes are closed to each other in the plurality of culture chambers by closing the communication holes.
When the plurality of culture chambers arranged in at least one direction by being divided by the dividing portion are stacked in the vertical direction,
The culture container according to any one of claims 1 to 4, wherein the plurality of culture chambers maintain a closed state.
A temperature maintenance device;
A culture container according to claim 1,
A pipe connected to the inlet / outlet of the culture container, a liquid injection part for injecting a liquid into the culture container through the pipe, and a liquid waste part for performing a waste liquid from the inside of the culture container through the pipe And an opening / closing part that opens and closes the entrance / exit,
A culture apparatus comprising a dividing drive unit for dividing the internal space into the dividing unit.

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