Material introduction device and material introduction unit

Abstract

To provide a substance introduction device and a substance introduction unit, which improve the usefulness of techniques for introducing substances into cells by electroporation.SOLUTION: A substance introduction device according to the present disclosure is a substance introduction device that introduces substances into cells by electroporation. The device comprises a housing portion made of a conductive material, which defines a storage space that can accommodate a cell suspension containing cells and substances; first and second electrode parts, which are capable of sandwiching the housing portion at opposing positions; and a voltage applying part for applying a voltage between the first electrode part and the second electrode part. At least one of the first and second electrode parts is equipped with a frame that presses the outer surface of the housing portion in a state in which the first and second electrode parts sandwich the housing portion to divide the housing space into a plurality of small chambers.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure relates to a substance introduction device and a substance introduction unit. [Background technology]

[0002] Conventionally, there are known techniques for obtaining a large amount of cells into which a substance such as DNA is introduced by electroporation. For example, Patent Document 1 discloses an electroporation device that applies a voltage to a number of wells that contain a cell suspension containing cells and a substance to be introduced into the cells, thereby carrying out electroporation. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2018-14916 A Summary of the Invention [Problem to be solved by the invention]

[0004] In electroporation, the optimal conditions for performing electroporation, such as the voltage value or application time of the voltage applied to the electrode pair, vary depending on environmental factors such as the amount of cell suspension to be treated in one go, the volume of the container in which the treatment is performed, the distance between the electrode pair to which the voltage is applied, etc. For this reason, even if the conditions for performing electroporation targeting a small amount of cells in a container such as a cuvette have been established, when performing electroporation in a different container to increase the amount of cell suspension to be treated in one go, it was necessary to reset the conditions for performing electroporation in the different container.

[0005] By increasing the number of wells in the device as in the device described in Patent Document 1, the total amount of cell suspension to be electroporated in one run can be increased under the same implementation conditions. On the other hand, since it is necessary to inject cell suspension into multiple wells or to recover cell suspension from multiple wells, the complexity of the work required to use the device increases with the increase in the number of wells. Therefore, there is a demand for further improvement in the usefulness of the technology for introducing substances into cells by electroporation.

[0006] In view of the above circumstances, an object of the present disclosure is to provide a substance introduction device and a substance introduction unit that improve the usefulness of the technique of introducing a substance into a cell by electroporation. [Means for solving the problem]

[0007] A substance introduction device according to one embodiment of the present disclosure is a substance introduction device that introduces a substance into cells by electroporation, and includes a storage section made of a conductive material that divides a storage space capable of containing the cells and a cell suspension containing the substance, a first electrode section and a second electrode section that can sandwich the storage section in opposing positions, and a voltage application section that applies a voltage between the first electrode section and the second electrode section, and at least one of the first electrode section and the second electrode section includes a frame that can press against an outer surface of the storage section when the first electrode section and the second electrode section are in a state where the storage section is sandwiched between the first electrode section and the second electrode section, thereby dividing the storage space into a plurality of small chambers.

[0008] In the substance introduction device according to an embodiment of the present disclosure, it is preferable that the frame be arranged in a lattice pattern in a plan view along the opposing direction of the first electrode unit and the second electrode unit.

[0009] In one embodiment of the substance introduction device of the present disclosure, it is preferable that the first electrode portion includes a plurality of first electrodes, the second electrode portion includes a plurality of second electrodes, and with the first electrode portion and the second electrode portion sandwiching the storage portion, each of the plurality of small chambers is sandwiched between any one of the plurality of first electrodes and any one of the plurality of second electrodes.

[0010] In the substance introduction device according to an embodiment of the present disclosure, the storage section is preferably in the form of a flexible bag.

[0011] In the substance introduction device according to one embodiment of the present disclosure, it is preferable that the container is removable from the substance introduction device.

[0012] In one embodiment of the substance introduction device of the present disclosure, the storage section preferably includes an injection port that determines the flow of the cell suspension in a direction in which the cell suspension is injected into the storage space, and a discharge port that determines the flow of the cell suspension in a direction in which the cell suspension is discharged from the storage space.

[0013] A substance introduction unit according to one embodiment of the present disclosure is a substance introduction unit used for introducing a substance into cells by electroporation, and comprises a container section made of a conductive material that divides a container space capable of containing the cells and a cell suspension containing the substance, and a first electrode section and a second electrode section that can sandwich the container section in opposing positions, and at least one of the first electrode section and the second electrode section has a frame that can press against an outer surface of the container section when the first electrode section and the second electrode section are in a state where the container section is sandwiched between them, thereby dividing the container space into a plurality of small chambers. Effect of the Invention

[0014] The substance introduction device and substance introduction unit according to the present disclosure can improve the usefulness of the technique of introducing a substance into a cell by electroporation. [Brief description of the drawings]

[0015] [Figure 1] 1 is a schematic diagram showing a schematic configuration of a substance introduction device according to an embodiment. [Diagram 2] 2 is a cross-sectional view taken along line II' of the substance introduction unit shown in FIG. 1. [Diagram 3] 3 is a cross-sectional view of the substance introduction unit at the same position as in FIG. 2, in a state in which the container is not sandwiched between the first electrode section and the second electrode section. FIG. [Figure 4] 2 is a plan view of the first electrode section shown in FIG. 1, as viewed along a direction perpendicular to a first electrode surface of the first electrode section. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Hereinafter, a substance introduction device 1 and a substance introduction unit 2 according to an embodiment of the present disclosure will be described with reference to the drawings.

[0017] In each drawing, the same or corresponding parts are denoted by the same reference numerals. In the description of this embodiment, the description of the same or corresponding parts will be omitted or simplified as appropriate.

[0018] The substance introduction device 1 according to the embodiment of the present disclosure introduces a substance into cells by electroporation. Electroporation is a method of applying an electric pulse or the like to a cell suspension containing cells and a substance to be introduced into the cells, thereby forming holes in the cell membrane of the cells and introducing the substance into the cells. Electroporation is also called electroporation. In this embodiment, the cell suspension is a suspension in a buffer solution of cells such as animal cells or E. coli, and substances such as genes, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), nucleic acids, proteins, low molecular weight compounds, lipid molecules, or liposomes.

[0019] A substance introduction device 1 according to an embodiment of the present disclosure will be described with reference to Fig. 1. Fig. 1 is a schematic diagram showing a schematic configuration of the substance introduction device 1. As shown in Fig. 1, the substance introduction device 1 includes a substance introduction unit 2, a voltage application section 3, a holding section 4, and a control section 5.

[0020] The substance introduction unit 2 is used for introducing a substance into a cell by electroporation. As shown in Fig. 2 and Fig. 3, the substance introduction unit 2 includes a first electrode section 21, a second electrode section 22, a container section 23, and a port 24. Fig. 2 is a cross-sectional view taken along line I-I' of the substance introduction unit 2 shown in Fig. 1. Fig. 3 is a cross-sectional view of the substance introduction unit 2 in the same position as Fig. 2, in a state in which the container section 23 is not sandwiched between the first electrode section 21 and the second electrode section 22.

[0021] As shown in FIG. 1, the substance introduction unit 2 is fixed to the substance introduction device 1 by being attached to the holding section 4 of the substance introduction device 1. Specifically, as shown by the arrow in FIG. 3, the holding section 4 presses the surface of the first electrode section 21 opposite the first electrode surface 21A and the surface of the second electrode section 22 opposite the second electrode surface 22A, so that the first electrode section 21 and the second electrode section 22 sandwich the container section 23 at a position where they face each other, as shown in FIG. 2. In this manner, the substance introduction unit 2 is attached to the holding section 4 of the substance introduction device 1. For this reason, in this embodiment, the substance introduction unit 2 is detachable from the substance introduction device 1, and further, the container section 23 is detachable from the substance introduction unit 2. As a result, after electroporation, the substance introduction unit 2 can be removed from the substance introduction device 1, and the container section 23 of the substance introduction unit 2 can be carried as a container that contains the introduced cell suspension in which the substance has been introduced into the cells by electroporation. Hereinafter, for convenience of explanation, the direction in which the first electrode section 21 and the second electrode section 22 face each other when the substance introduction unit 2 is fixed to the substance introduction device 1 may be referred to as the facing direction of the first electrode section 21 and the second electrode section 22, or simply as the "facing direction." However, at least one of the first electrode section 21, the second electrode section 22, the storage section 23, and the port 24 constituting the substance introduction unit 2 may be formed integrally with the substance introduction device 1 and may not be removable from the substance introduction device 1.

[0022] The first electrode section 21 includes a plurality of first electrodes 211. Specifically, as shown in FIG. 4, the first electrode section 21 includes a rectangular plate-shaped electrode holding section 21B and a plurality of rectangular plate-shaped first electrodes 211 arranged side by side on the electrode holding section 21B to form a flat first electrode surface 21A as a whole. In other words, the first electrode surface 21A is formed by the electrode surfaces of the plurality of first electrodes 211 arranged in the same plane. FIG. 4 is a plan view of the first electrode section 21 shown in FIG. 1 as viewed in a direction perpendicular to the first electrode surface 21A of the first electrode section 21. In this embodiment, 35 electrodes are arranged on the electrode holding section 21B as the plurality of first electrodes 211 in a two-dimensional array of 5×7. However, the number of first electrodes 211 included in the first electrode unit 21 may be any natural number equal to or greater than 2, and the first electrodes 211 may configure the first electrode surface 21A in any arrangement, such as a one-dimensional array or a two-dimensional array. Furthermore, the shape of the electrode holding unit 21B and the first electrodes 211 of the first electrode unit 21 is not limited to a plate shape, and may be any shape that allows the first electrode surface 21A to be configured as a whole of the multiple first electrodes 211. The first electrodes 211 are formed of a material that can conduct electricity, such as a metal or a conductive polymer. Furthermore, the parts of the first electrode unit 21 other than the first electrodes 211, such as the electrode holding unit 21B, may be formed of an insulating material, such as plastic, glass, silicon, or quartz.

[0023] The first electrode section 21 includes a frame 212. As shown in FIG. 2 and FIG. 3, the frame 212 of the first electrode section 21 can press the outer surface of the housing section 23 to divide the housing space 23A of the housing section 23 into a plurality of small chambers 23B when the first electrode section 21 and the second electrode section 22 sandwich the housing section 23. Specifically, the frame 212 protrudes from the first electrode surface 21A of the first electrode section 21 in a direction perpendicular to the first electrode surface 21A. More specifically, the frame 212 protrudes from the electrode holding section 21B at a position between the plurality of first electrodes 211. The frame 212 also protrudes from the first electrode surface 21A formed by the plurality of first electrodes 211. The frame 212 may be molded integrally with the electrode holding section 21B, or may be fixed to the electrode holding section 21B by adhesion or the like. In addition, the frame 212 may be detachable from the electrode holding portion 21B without being bonded to the electrode holding portion 21B. When the frame 212 is detachable from the electrode holding portion 21B, the frame 212 may be replaced depending on the capacity or shape of the small chamber 23B to be partitioned in the storage portion 23. As shown in FIG. 4, the frame 212 may be arranged in a lattice shape in a plan view seen along a direction perpendicular to the first electrode surface 21A of the first electrode portion 21, that is, in a plan view seen along the opposing direction of the first electrode portion 21 and the second electrode portion 22. This makes it possible to uniform the capacity or shape of the small chamber 23B partitioned in the storage portion 23, and the frame 212 of the first electrode portion 21 can apply a force evenly to the outer surface of the storage portion 23 with the first electrode portion 21 and the second electrode portion 22 sandwiching the storage portion 23. In this embodiment, the protruding amount of the frame 212 is approximately constant regardless of the position in the in-plane direction of the first electrode surface 21A. In this embodiment, the lattice-shaped frame 212 is arranged so as to surround each of the multiple first electrodes 211 arranged on the first electrode surface 21A in a plan view along the opposing direction of the first electrode portion 21 and the second electrode portion 22. The frame 212 may be formed of an insulating material such as plastic, glass, silicon, or quartz. The frame 212 may be provided with a concave cutout so as to avoid the port 24 when the first electrode portion 21 and the second electrode portion 22 sandwich the accommodation portion 23.

[0024] The second electrode section 22 includes a plurality of second electrodes 221. In this embodiment, the second electrode section 22 has the same shape as the first electrode section 21. Specifically, the second electrode section 22 includes a rectangular plate-shaped electrode holding section 22B and a plurality of rectangular plate-shaped second electrodes 221 arranged side by side on the electrode holding section 22B to form a flat second electrode surface 22A as a whole. In other words, the second electrode surface 22A is formed by the electrode surfaces of the plurality of second electrodes 221 arranged in the same plane. In this embodiment, 35 electrodes are arranged on the electrode holding section 22B as the plurality of second electrodes 221 in a two-dimensional array of 5×7. However, the number of second electrodes 221 included in the second electrode section 22 may be any natural number equal to or greater than 2, and the second electrodes 221 may form the second electrode surface 22A in any arrangement, such as a one-dimensional array or a two-dimensional array. Moreover, the shape of the electrode holding portion 22B and the second electrode 221 of the second electrode portion 22 is not limited to a plate shape, and may be any shape that allows the second electrode surface 22A to be configured by the entirety of the multiple second electrodes 221. The second electrode 221 is formed of a material that can conduct electricity, such as a metal or a conductive polymer. Furthermore, the portions of the second electrode portion 22 other than the second electrode 221, such as the electrode holding portion 22B, may be formed of an insulating material, such as plastic, glass, silicon, or quartz. In this embodiment, the first electrode portion 21 and the second electrode portion 22 are assumed to have the same shape, but this is not limited thereto.

[0025] The second electrode section 22 includes a frame 222. As shown in FIG. 2 and FIG. 3, the frame 212 of the second electrode section 22 can press the outer surface of the housing section 23 to divide the housing space 23A of the housing section 23 into a plurality of small chambers 23B when the first electrode section 21 and the second electrode section 22 sandwich the housing section 23. Specifically, the frame 222 protrudes from the second electrode surface 22A of the second electrode section 22 in a direction perpendicular to the second electrode surface 22A. More specifically, the frame 222 protrudes from the electrode holding section 22B at a position between the plurality of second electrodes 221. The frame 222 protrudes from the second electrode surface 22A formed by the plurality of second electrodes 221. The frame 222 may be molded integrally with the electrode holding section 22B, or may be fixed to the electrode holding section 22B by adhesion or the like. In addition, the frame 222 may be detachable from the electrode holding portion 22B without being bonded to the electrode holding portion 22B. When the frame 222 is detachable from the electrode holding portion 22B, the frame 222 may be replaced depending on the capacity or shape of the small chamber 23B to be partitioned in the storage portion 23. As with the frame 212 of the first electrode portion 21, the frame 222 may be arranged in a lattice shape in a plan view seen along a direction perpendicular to the second electrode surface 22A of the second electrode portion 22, that is, in a plan view seen along the opposing direction of the first electrode portion 21 and the second electrode portion 22. This makes it possible to make the capacity or shape of the small chamber 23B partitioned in the storage portion 23 uniform, and allows the frame 222 of the second electrode portion 22 to apply a force uniformly to the outer surface of the storage portion 23 with the first electrode portion 21 and the second electrode portion 22 sandwiching the storage portion 23. In this embodiment, the amount of protrusion of the frame 222 is approximately constant regardless of the position in the in-plane direction of the second electrode surface 22A. In this embodiment, the lattice-shaped frame 222 is arranged so as to surround each of the multiple second electrodes 221 arranged on the second electrode surface 22A in a plan view seen along the opposing direction of the first electrode portion 21 and the second electrode portion 22. The frame 222 may be formed of an insulating material such as plastic, glass, silicon, or quartz. The frame 222 may be provided with a concave cutout so as to avoid the port 24 when the first electrode portion 21 and the second electrode portion 22 sandwich the accommodation portion 23.

[0026] The storage unit 23 is configured to store a cell suspension containing cells and a substance to be introduced into the cells. As shown in Fig. 3, the storage unit 23 defines a storage space 23A capable of storing the cell suspension. The storage unit 23 is a container formed of a conductive material such as a conductive polymer. The capacity of the storage unit 23 is determined arbitrarily according to the amount of the cell suspension to be treated in one electroporation, and is, for example, 10 milliliters to 100 milliliters.

[0027] The accommodation portion 23 may be a flexible bag-shaped container. The accommodation portion 23 is made of a material softer than the material of the first electrode portion 21 and the second electrode portion 22 so that the shape of the accommodation portion 23 deforms along the shapes of the first electrode portion 21 and the second electrode portion 22 by being sandwiched between the first electrode portion 21 and the second electrode portion 22.

[0028] 2, in a state in which the first electrode section 21 and the second electrode section 22 sandwich the housing section 23, the frame 212 of the first electrode section 21 and the frame 222 of the second electrode section 22 can press the outer surface of the housing section 23 to divide the housing space 23A of the housing section 23 into a plurality of small chambers 23B. At this time, since the housing section 23 has flexibility, in a state in which the housing section 23 is sandwiched between the first electrode section 21 and the second electrode section 22, a gap is unlikely to be generated between the outer surface of the housing section 23 and each of the first electrode section 21 and the second electrode section 22. Then, in a state in which the housing section 23 is sandwiched between the first electrode section 21 and the second electrode section 22, each of the plurality of small chambers 23B is sandwiched between any one of the plurality of first electrodes 211 and any one of the plurality of second electrodes 221.

[0029] At this time, by making conditions such as the capacity of the small chamber 23B and the size and distance of the first electrode 211 and the second electrode 221 sandwiching the small chamber 23B the same as the conditions for performing electroporation for a small amount of cells established in a previous experiment using a container such as a cuvette, it is possible to increase the amount of cell suspension to be processed at one time according to the number of small chambers 23B partitioned in the storage section 23 without resetting the conditions for performing electroporation. The capacity of the small chamber 23B is arbitrarily determined according to the amount of cell suspension to be processed at one electroporation in a previous experiment, and is, for example, 10 microliters to 10 milliliters, and preferably 100 microliters to 1 milliliter.

[0030] In this embodiment, the frame 212 of the first electrode unit 21 and the frame 222 of the second electrode unit 22 can sandwich the housing unit 23 so that the outer surface of the housing unit 23 on the first electrode unit 21 side and the outer surface of the housing unit 23 on the second electrode unit 22 side are in contact with each other in the opposing direction of the first electrode unit 21 and the second electrode unit 22, in other words, the housing unit 23 is divided. This makes it difficult for a dead volume to occur between the frame 212 of the first electrode unit 21 and the frame 222 of the second electrode unit 22, where an electric pulse does not reach, during electroporation. On the other hand, the frame 212 of the first electrode unit 21 and the frame 222 of the second electrode unit 22 may sandwich the housing unit 23 so that the outer surface of the housing unit 23 on the first electrode unit 21 side and the outer surface of the housing unit 23 on the second electrode unit 22 side are not in contact with each other in the opposing direction of the first electrode unit 21 and the second electrode unit 22, in other words, the housing space 23A is not completely divided. This can prevent the housing portion 23 from being damaged due to an excessive load being applied to the outer surface of the housing portion 23.

[0031] In addition, in this embodiment, the first electrode portion 21 and the second electrode portion 22 are provided with the frames 212 and 222, respectively, but this is not limited thereto. Either the first electrode portion 21 or the second electrode portion 22 may be provided with the frame 212 or 222. For example, only the first electrode portion 21 may be provided with the frame 212. In such a case, the outer surface of the accommodation portion 23 is sandwiched between the frame 212 of the first electrode portion 21 and the electrode holding portion 22B of the second electrode portion 22, so that the accommodation space 23A of the accommodation portion 23 can be divided into a plurality of small chambers 23B. Similarly, only the second electrode portion 22 may be provided with the frame 222.

[0032] The port 24 is a cylindrical member capable of injecting a cell suspension into the storage space 23A of the storage unit 23 or discharging the cell suspension from the storage space 23A of the storage unit 23. The port 24 is equipped with, for example, an open / close valve. In this case, by opening the valve of the port 24, it becomes possible to inject the cell suspension into the storage space 23A of the storage unit 23 or to discharge the cell suspension from the storage space 23A of the storage unit 23.

[0033] The port 24 may include an injection port 24A and a discharge port 24B. The injection port 24A and the discharge port 24B each include a check valve or the like, and regulate the flow of the cell suspension flowing therethrough. The injection port 24A regulates the flow of the cell suspension in a direction in which the cell suspension is injected into the accommodation space 23A of the accommodation section 23. The injection port 24A may be connected to the pre-introduction bag 10, which contains the cell suspension before electroporation, by a liquid supply tube. The discharge port 24B regulates the flow of the cell suspension in a direction in which the cell suspension is discharged from the accommodation space 23A of the accommodation section 23. The discharge port 24B may be connected to the introduced bag 11, which contains the cell suspension after electroporation, by a liquid supply tube. This facilitates the injection and discharge of the cell suspension into and from the accommodation space 23A of the accommodation section 23, and further improves the convenience of the substance introduction unit 2. In this embodiment, the injection port 24A and the discharge port 24B are provided at positions on both sides of the storage unit 23 in a plan view along the opposing direction of the first electrode unit 21 and the second electrode unit 22. Note that, in this embodiment, a cell suspension in which cells and a substance to be introduced into the cells are suspended in a buffer solution in advance is used, but this is not limited thereto. For example, a cell suspension containing cells and a substance to be introduced into the cells may be mixed at the timing when they are poured into the storage space 23A of the storage unit 23, or may be poured into the storage space 23A of the storage unit 23 at different times and mixed in the storage space 23A of the storage unit 23.

[0034] Referring again to FIG. 1, the voltage application unit 3 applies a voltage between the first electrode unit 21 and the second electrode unit 22. The voltage application unit 3 may include, for example, a storage battery or a dry cell. The voltage application unit 3 may also include, for example, an adapter for receiving power from an external power source, and may be configured to receive power from the external power source. As shown by solid lines in FIG. 1, the voltage application unit 3 is connected to each of the first electrode unit 21 and the second electrode unit 22 by a conductor or the like so as to be able to supply power. More specifically, the voltage application unit 3 applies a voltage between pairs of the first electrode 211 and the second electrode 221 that sandwich each of the multiple small chambers 23B partitioned in the storage unit 23. The voltage application unit 3 may be configured to sequentially apply a voltage to the multiple pairs of the first electrode 211 and the second electrode 221, or may be configured to simultaneously apply a voltage. When the voltage application unit 3 applies a predetermined voltage between the pair of first electrode 211 and second electrode 221 that sandwich each of the multiple chambers 23B, a voltage is applied to the cell suspension contained in the chambers 23B, and a substance is introduced into the cells by electroporation. In this way, since electroporation can be performed for each cell suspension contained in the chambers 23B partitioned in the storage unit 23, the amount of cell suspension to be processed at one time can be increased by increasing the number of chambers 23B without changing the conditions for performing electroporation.

[0035] The holding section 4 holds the substance introduction unit 2. In this embodiment, the holding section 4 includes a first holding member 41 and a second holding member 42. In the holding section 4, one of the first holding member 41 and the second holding member 42 supports one of the first electrode section 21 and the second electrode section 22. This allows the substance introduction unit 2 to be held by the holding section 4. In this embodiment, the second holding member 42 is configured to support the second electrode section 22 of the substance introduction unit 2 from the surface side opposite to the second electrode surface 22A. That is, in this embodiment, the substance introduction unit 2 is supported by the second holding member 42 of the holding section 4, and thereby held by the holding section 4. The holder 4 can sandwich the substance introduction unit 2 by pressing the first electrode section 21 from the surface opposite to the first electrode surface 21A of the first electrode section 21 with the first holding member 41, and pressing the second electrode section 22 from the surface opposite to the second electrode surface 22A of the second electrode section 22 with the second holding member 42. However, the holder 4 may have any structure capable of holding the substance introduction unit 2. The holder 4 is formed of an insulating material such as plastic, glass, silicon, or quartz.

[0036] The control unit 5 includes, for example, a memory 51 and a processor 52. As the control unit 5, for example, an electroporator, a personal computer, a smartphone, a tablet terminal, or other such computer may be used.

[0037] The memory 51 is, for example, a semiconductor memory, a magnetic memory, or an optical memory. The memory 51 functions as, for example, a main storage device, an auxiliary storage device, or a cache memory. The memory 51 stores any information used in the operation of the substance introduction device 1. For example, the memory 51 stores a system program, an application program, or embedded software.

[0038] The processor 52 may be, for example, a general-purpose processor such as a CPU (Central Processing Unit), or a dedicated processor specialized for a specific process, etc. The processor 52 may include, for example, a dedicated circuit such as a FPGA (Field-Programmable Gate Array) or an ASIC (Application Specific Integrated Circuit).

[0039] 1, the control unit 5 is connected to the voltage application unit 3 so as to be able to communicate with each other via wire or wirelessly. In this way, the control unit 5 controls components such as the voltage application unit 3 to realize the functions of the substance introduction device 1. Specifically, the control unit 5 can control the voltage application unit 3 to apply a predetermined voltage between the first electrode unit 21 and the second electrode unit 22.

[0040] An example of the operation of the substance introduction device 1 according to this embodiment will be described below. First, the storage unit 23 of the substance introduction device 1 is connected via the injection port 24A to a pre-introduction bag 10 that contains a cell suspension before electroporation, and is connected via the discharge port 24B to a post-introduction bag 11 that contains a cell suspension after electroporation.

[0041] The cell suspension is injected from the pre-introduction bag 10 through the injection port 24A into the storage space 23A of the storage unit 23. As a result, the storage unit 23 stores the cell suspension containing the cells and the substance in the internal storage space 23A. After the cell suspension is injected into the storage unit 23, the injection port 24A is closed.

[0042] The substance introduction unit 2 is attached to the holding part 4 of the substance introduction device 1 in a state in which the first electrode part 21 and the second electrode part 22 are positioned opposite to each other and sandwich the accommodation part 23. As a result, the first electrode part 21 and the second electrode part 22 sandwich the accommodation part 23, and the frame 212 of the first electrode part 21 and the frame 222 of the second electrode part 22 press the outer surface of the accommodation part 23 to divide the accommodation space 23A into a plurality of small chambers 23B.

[0043] The voltage application unit 3 applies a voltage between the first electrode unit 21 and the second electrode unit 22. When the voltage application unit 3 applies a predetermined voltage between a pair of a first electrode 211 and a second electrode 221 that sandwich each of a plurality of small chambers 23B partitioned in the storage unit 23, a voltage is applied to the cell suspension contained in the small chambers 23B, and a substance is introduced into the cells by electroporation.

[0044] After electroporation, the substance introduction unit 2 is removed from the holding part 4 of the substance introduction device 1. This releases the container part 23 from the first electrode part 21 and the second electrode part 22.

[0045] The cell suspension is discharged from the storage space 23A of the storage unit 23 through the discharge port 24B into the introduced bag 11. At that time, air or a buffer solution may be injected into the storage space 23A from the injection port 24A. This allows the cell suspension in the storage space 23A of the storage unit 23 to be collected without remaining therein. Alternatively, the storage unit 23 may be removed from the substance introduction device 1 and used as a container for storing the introduced cell suspension.

[0046] As described above, the substance introduction device 1 according to the present disclosure is a substance introduction device 1 that introduces a substance into cells by electroporation. The substance introduction device 1 includes a container 23 formed of a conductive material that defines a container space 23A capable of containing a cell suspension containing cells and a substance, a first electrode section 21 and a second electrode section 22 that can sandwich the container 23 at opposing positions, and a voltage application section 3 that applies a voltage between the first electrode section 21 and the second electrode section 22. At least one of the first electrode section 21 and the second electrode section 22 includes a frame 212 or 222 that can press the outer surface of the container 23 to divide the container space 23A into a plurality of small chambers 23B when the first electrode section 21 and the second electrode section 22 sandwich the container 23. With this configuration, the substance introduction device 1 can perform electroporation for each cell suspension contained in the small chambers 23B defined in the container 23, and therefore the amount of cell suspension to be processed at one time can be increased without changing the conditions for performing electroporation. Furthermore, the substance introduction device 1 can divide the accommodation space 23A of the accommodation unit 23 into a plurality of small chambers 23B by sandwiching the accommodation unit 23 between the first electrode unit 21 and the second electrode unit 22, and can return the accommodation unit 23 to its original accommodation space 23A by opening the accommodation unit 23 from the first electrode unit 21 and the second electrode unit 22. This eliminates the need to perform complicated operations such as preparing a plurality of containers to which the same electroporation conditions can be applied, injecting a cell suspension into each container, or recovering a cell suspension from each container. Therefore, the substance introduction device 1 can improve the usefulness of the technique of introducing a substance into cells by electroporation.

[0047] In the substance introduction device 1 according to the present disclosure, the frames 212 or 222 are preferably arranged in a lattice pattern in a plan view along the opposing direction of the first electrode unit 21 and the second electrode unit 22. With such a configuration, the substance introduction device 1 can make the capacity or shape of the small chambers 23B partitioned in the storage unit 23 uniform, and can allow the frame 212 of the first electrode unit 21 to apply a force uniformly to the outer surface of the storage unit 23 with the first electrode unit 21 and the second electrode unit 22 sandwiching the storage unit 23.

[0048] In the substance introduction device 1 according to the present disclosure, the first electrode section 21 includes a plurality of first electrodes 211, the second electrode section 22 includes a plurality of second electrodes 221, and it is preferable that each of the plurality of chambers 23B is sandwiched between any one of the plurality of first electrodes 211 and any one of the plurality of second electrodes 221 in a state in which the first electrode section 21 and the second electrode section 22 sandwich the storage section 23. According to this configuration, the substance introduction device 1 can perform electroporation by applying a voltage to each of the cell suspensions contained in the plurality of chambers 23B individually. For example, the substance introduction device 1 may apply a voltage to all of the plurality of chambers 23B simultaneously, or may apply a voltage to the plurality of chambers 23B sequentially.

[0049] In the substance introduction device 1 according to the present disclosure, the storage section 23 is preferably in the form of a flexible bag. With this configuration, when the first electrode section 21 and the second electrode section 22 sandwich the storage section 23 and press the outer surface of the storage section 23 to divide the storage space 23A into a plurality of small chambers 23B, gaps are unlikely to be generated between the outer surface of the storage section 23 and each of the first electrode section 21 and the second electrode section 22.

[0050] In the substance introduction device 1 according to the present disclosure, the storage section 23 is preferably removable from the substance introduction device 1. With this configuration, after electroporation, the storage section 23 can be removed from the substance introduction device 1 and the substance introduction unit 2 and carried as a container that contains the introduced cell suspension.

[0051] In the substance introduction device 1 according to the present disclosure, the storage unit 23 preferably includes an injection port that regulates the flow of the cell suspension in a direction in which the cell suspension is injected into the storage space 23A, and a discharge port that regulates the flow of the cell suspension in a direction in which the cell suspension is discharged from the storage space 23A. This configuration facilitates the injection and discharge of the cell suspension into and from the storage space 23A of the storage unit 23, further improving the convenience of the substance introduction device 1.

[0052] The substance introduction unit 2 according to the present disclosure is a substance introduction unit 2 used for introducing a substance into cells by electroporation. The substance introduction unit 2 includes a container 23 formed of a conductive material that defines a container space 23A capable of containing a cell suspension containing cells and a substance, and a first electrode section 21 and a second electrode section 22 that can sandwich the container 23 at opposing positions. At least one of the first electrode section 21 and the second electrode section 22 includes a frame 212 or 222 that can press the outer surface of the container 23 to divide the container space 23A into a plurality of small chambers 23B when the first electrode section 21 and the second electrode section 22 are in a state in which the container 23 is sandwiched between them. With this configuration, the substance introduction unit 2 can perform electroporation for each cell suspension contained in the small chambers 23B defined in the container 23, and therefore the amount of cell suspension to be processed at one time can be increased without changing the conditions for performing electroporation. Furthermore, the substance introduction unit 2 divides the accommodation space 23A of the accommodation section 23 into a plurality of small chambers 23B by sandwiching the accommodation section 23 between the first electrode section 21 and the second electrode section 22, and by opening the accommodation section 23 from the first electrode section 21 and the second electrode section 22, the accommodation space 23A of the accommodation section 23 can be returned to its original state. This eliminates the need to perform complicated operations such as preparing a plurality of containers to which the same electroporation conditions can be applied, injecting a cell suspension into each container, or recovering a cell suspension from each container. Therefore, the substance introduction unit 2 can improve the usefulness of the technique of introducing a substance into cells by electroporation.

[0053] Although the present disclosure has been described based on the drawings and embodiments, it should be noted that those skilled in the art can make various modifications and corrections based on the present disclosure. Therefore, it should be noted that these modifications and corrections are included in the scope of the present disclosure. For example, the functions included in each means or step can be rearranged so as not to be logically inconsistent, and multiple means or steps can be combined or divided into one.

[0054] For example, in the above-described embodiment, the substance introduction device 1 has been described as including the control unit 5, and the voltage application unit 3 is controlled by the control unit 5. However, the substance introduction device 1 may not include the control unit 5, and the voltage application unit 3 may be controlled manually. This reduces the manufacturing cost of the substance introduction device 1, and the substance introduction device 1 can be provided at low cost.

[0055] Alternatively, in the above-mentioned embodiment, the number of the first electrodes 211 and the second electrodes 221 included in the substance introduction unit 2 has been described as being the same as the number of the small chambers 23B. However, the number of the first electrodes 211 and the second electrodes 221 may be different from the number of the small chambers 23B. For example, in a state in which the container 23 is sandwiched between the first electrode section 21 and the second electrode section 22, a plurality of small chambers 23B may be sandwiched between one each of the first electrode 211 and the second electrode section 22. Furthermore, instead of the first electrode section 21 including a plurality of first electrodes 211 and the second electrode section 22 including a plurality of second electrodes 221, the first electrode section 21 and the second electrode section 22 may each include one large first electrode 211 or one large second electrode 221. [Explanation of symbols]

[0056] 1 Substance introduction device 10 Pre-introduction bag 11 Introduced Bags 2. Material Introduction Unit 21 1st electrode section 21A electrode surface 21B Electrode holding part 211 1st electrode 212 frames 22 Second electrode part 22A electrode surface 22B Electrode holding part 221 2nd electrode 222 frames 23 Storage unit 23A Storage Space 23B Small room 24 ports 24A Injection Port 24B Discharge port 3. Voltage application section 4 Holding part 41 First holding member 42 Second holding member 5. Control section 51 Memory 52 processors

Claims

1. A substance introduction device for introducing a substance into a cell by electroporation, comprising: A storage section formed of a conductive material, which defines a storage space capable of storing a cell suspension containing the cells and the substance; A first electrode portion and a second electrode portion capable of sandwiching the housing portion at opposing positions; a voltage application unit that applies a voltage between the first electrode unit and the second electrode unit, At least one of the first electrode portion and the second electrode portion includes a frame that can press an outer surface of the storage portion to divide the storage space into a plurality of small chambers when the first electrode portion and the second electrode portion sandwich the storage portion, The substance introduction device, wherein the container is in the form of a flexible bag.

2. The substance introduction device according to claim 1 , wherein the frame is arranged in a lattice pattern in a plan view along a direction in which the first electrode unit and the second electrode unit face each other.

3. The first electrode unit includes a plurality of first electrodes, The second electrode portion includes a plurality of second electrodes, The substance introduction device according to claim 1 or 2, wherein, when the first electrode portion and the second electrode portion sandwich the storage portion, each of the multiple chambers is sandwiched between any one of the multiple first electrodes and any one of the multiple second electrodes.

4. The substance introduction device according to claim 1 , wherein the container is removable from the substance introduction device.

5. The storage section includes: an injection port that defines a flow of the cell suspension in a direction in which the cell suspension is injected into the accommodation space; a discharge port that defines a flow of the cell suspension in a direction in which the cell suspension is discharged from the storage space; The substance introduction device according to claim 1 , comprising:

6. A substance introduction unit used for introducing a substance into a cell by electroporation, comprising: A storage section formed of a conductive material, which defines a storage space capable of storing a cell suspension containing the cells and the substance; A first electrode portion and a second electrode portion capable of sandwiching the storage portion at opposing positions, At least one of the first electrode portion and the second electrode portion includes a frame that can press an outer surface of the storage portion to divide the storage space into a plurality of small chambers when the first electrode portion and the second electrode portion sandwich the storage portion, The container is in the form of a flexible bag.

7. When the first electrode portion and the second electrode portion sandwich the storage portion, the frame brings the inner surface of the storage portion on the first electrode portion side into contact with the inner surface of the storage portion on the second electrode portion side, completely dividing the storage space and dividing the multiple small chambers, each of which is surrounded by the inner surface of the storage portion; the accommodation portion changes shape along shapes of the first electrode portion and the second electrode portion when the first electrode portion and the second electrode portion sandwich the accommodation portion, and comes into contact with each of the first electrode portion and the second electrode portion such that no gap is generated between the outer surface of the accommodation portion and each of the first electrode portion and the second electrode portion, The substance introduction device according to claim 2 , wherein when the storage section is released from being sandwiched between the first electrode section and the second electrode section, the storage section defines the storage space that was the same as the space before being sandwiched between the first electrode section and the second electrode section.

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