Container for plasma-type cell treatment apparatus, and cell treatment method

Abstract

Provided is a container for a plasma-type cell treatment apparatus that treats an object of interest with plasma, the container comprising: a container body; an electrode that is installed in an upper portion of the inside of the container body and generates plasma; a treatment unit that is provided in a lower portion of the inside of the container body; a flow path that allows the inside and outside of the container body to communicate with each other; and a variable mechanism that changes the relative position between the electrode and the treatment unit. The variable mechanism preferably has a connection part for connecting the container body to the electrode. The variable mechanism is preferably provided on a side surface of the container body.

Description

Container for plasma cell processing device and cell processing method

[0001] The present invention relates to a container for a plasma cell treatment device and a cell treatment method.This application claims priority based on Japanese Patent Application No. 2024-148298, filed on August 30, 2024, the contents of which are incorporated herein by reference.

[0002] In genetic engineering and pharmaceutical development, molecules are introduced into target cells or target tissues (hereinafter collectively referred to as "target cells, etc.") to test the function and physiological activity of the molecules in the target cells, etc. Examples of molecules include polynucleotides, signaling proteins, transcriptional regulators, small molecule physiologically active substances, and drug candidates.

[0003] Methods for introducing molecules include electroporation, gene gun, liposome, cell fusion, viral vector, etc. However, the efficiency of introducing molecules into target cells and the like using these molecule introduction methods has not yet been satisfactory.

[0004] To address these problems, Patent Documents 1 and 2 propose a plasma cell treatment device that includes a capillary electrode, a counter electrode provided on the opposite side of the capillary electrode, and a power supply connected to the capillary electrode and the counter electrode. According to the inventions of Patent Documents 1 and 2, plasma generated between the electrodes stimulates target cells, which are the object of treatment, and introduces molecules into the target cells.

[0005] Patent No. 6189019 Patent No. 5737828

[0006] The inventions of Patent Documents 1 and 2 involve discharging a plasma treatment electrode placed on the upper surface of a cell culture surface, so that the target cells or other objects to be treated are exposed to the treatment chamber or space during treatment. If the treatment chamber or treatment space is exposed to the target cells or other objects, contamination with the target cells or other objects may occur. The present invention aims to provide a container for a plasma cell treatment device that can treat objects to be treated without exposing them to the external atmosphere.

[0007] The present invention has the following aspects. <1> A container for a plasma cell processing device that processes an object to be processed with plasma, the container for the plasma cell processing device comprising: a container body; an electrode that is installed in the upper part of the container body and generates plasma; a processing unit that is installed in the lower part of the container body; a flow path that connects the inside and outside of the container body; and an adjustable mechanism that changes the relative position between the electrode and the processing unit. <2> The container for the plasma cell processing device according to <1>, wherein the adjustable mechanism has a connector that connects the container body and the electrode. <3> The container for the plasma cell processing device according to <1>, wherein the adjustable mechanism is installed on a side surface of the container body. <4> The container for the plasma cell processing device according to any one of <1> to <3>, wherein the adjustable mechanism is configured so that the distance between the electrode and the object to be processed in the container body can be changed to 0.1 to 5 mm by the adjustable mechanism. <5> The container for the plasma cell processing device according to any one of <1> to <4>, wherein the container body is composed of a plurality of parts that are detachable from each other. <6> A container for a plasma cell processing device according to any one of <1> to <5>, wherein the electrode is a needle electrode. <7> A container for a plasma cell processing device according to any one of <1> to <6>, wherein the electrode has a partition wall made of a dielectric at its lower end. <8> A cell processing method using a plasma cell processing device equipped with the container according to any one of <1> to <7>, comprising: a pre-culture step of culturing a material to be processed in a culture solution within the container; a discharge step of discharging the culture solution from the container; a contact step of bringing molecules into contact with the material to be processed within the container after the discharge step; and an irradiation step of irradiating the material to be processed with plasma after the contact step, wherein the position of the electrode in the irradiation step is lower than the position of the electrode in the pre-culture step.

[0008] The present invention further has the following aspects. <1A> A container for a plasma cell processing device used in a plasma cell processing device that stimulates a treatment target with plasma generated between spaced electrodes, the container having a container with a storage chamber inside and an opening at an upper end, a lid that closes the opening, and a connector that connects to one of the spaced electrodes, the container for a plasma cell processing device further having an adjustable mechanism that changes the distance between the tip of the connector and the inner bottom surface of the container within the storage chamber when the opening is closed with the lid. <2A> The container for a plasma cell processing device according to <1A>, having a flow path that connects the inside and outside of the storage chamber. <3A> The container for a plasma cell processing device according to <1A> or <2A>, the connector is provided on the lid, the connector extends toward the inner bottom surface of the container, and the adjustable mechanism displaces the connector up and down relative to the lid. <4A> A container for a plasma cell processing device according to <1A> or <2A>, wherein the connecting portion is provided on the lid, the connecting portion extends toward the inner bottom surface of the container, and the variable mechanism displaces the lid up and down relative to the container. <5A> A container for a plasma cell processing device according to any of <1A> to <4A>, wherein the plasma cell processing device is for introducing molecules. <6A> A plasma cell processing device comprising a container for a plasma cell processing device according to any of <1A> to <4A> and spaced electrodes, wherein one of the spaced electrodes is connected to the connecting portion, and the other of the spaced electrodes is located outside the container. <7A> A plasma cell processing device according to <6A>, wherein the plasma cell processing device is for introducing molecules. <8A> A molecule introduction method using the plasma cell treatment device described in <6A>, comprising: a contact step of bringing a molecule into contact with a target cell or target tissue in the storage chamber; and an irradiation step of irradiating the molecule with plasma while bringing the molecule into contact with the target cell or target tissue in the storage chamber.<9A> A molecule introduction method using the plasma cell processing device described in <6A>, comprising: a pre-culture step of culturing target cells or target tissues in a culture solution within the storage chamber; a contact step of bringing molecules into contact with the target cells or the target tissue within the storage chamber after the pre-culture step; and an irradiation step of irradiating the target cells or the target tissue with plasma while contacting the molecules with the target cells or the target tissue within the storage chamber, wherein the pre-culture step cultures the target cells or the target tissue with a tip of the connection part positioned so as not to contact the target cells, the target tissue, or the culture solution, and then discharges the culture solution from the storage chamber, and the irradiation step irradiates the plasma while the opening is closed with the lid.

[0009] According to the container for the plasma cell treatment device of the present invention, the treatment object can be treated without being exposed to the external atmosphere.

[0010] FIG. 1 is a cross-sectional view of a plasma cell processing device according to a first embodiment. FIG. 1 is a cross-sectional view of a plasma cell processing device according to a first embodiment. FIG. 2 is a cross-sectional view of a plasma cell processing device according to a first embodiment. FIG. 3 is a cross-sectional view of a plasma cell processing device according to a first embodiment. FIG. 4 is a cross-sectional view of a plasma cell processing device according to a second embodiment. FIG. 5 is a cross-sectional view of a plasma cell processing device according to a third embodiment. FIG. 6 is a cross-sectional view of a plasma cell processing device according to one embodiment. FIG. 7 is a cross-sectional view of a plasma cell processing device according to a fourth embodiment. FIG. 8 is a cross-sectional view of a plasma cell processing device according to a fifth embodiment. FIG. 9 is a cross-sectional view of a plasma cell processing device according to a sixth embodiment. FIG. 10 is a cross-sectional view of a plasma cell processing device according to a seventh embodiment. FIG. 11 is a cross-sectional view of a plasma cell processing device according to the first embodiment.

[0011] In this specification and claims, the use of "to" indicating a range of values ​​means that the values ​​before and after it are included as the lower and upper limits.

[0012] The container for the plasma cell processing device of the present invention (hereinafter sometimes referred to as the "cell processing device") comprises a container body, an electrode installed in the upper part of the container body to generate plasma, a processing section installed in the lower part of the container body, a flow path connecting the inside and outside of the container body, and a variable mechanism for changing the relative position between the electrode and the processing section. The cell processing device stimulates a processing object (subject to be processed), such as a target cell or target tissue (collectively referred to as "target cells, etc."). For example, the cell processing device generates plasma between spaced electrodes, irradiates an irradiation object in which the target cells, etc. and molecules coexist, and introduces molecules into the processing object, such as the target cells, etc. The introduction of molecules proceeds primarily by inducing endocytosis using plasma. The cell processing device of the present invention will be described below with reference to an embodiment.

[0013] First Embodiment <Cell Processing Apparatus> The cell processing apparatus 1 in FIGS. 1 to 3 has a first electrode 10, a second electrode 20, a power supply unit 30, and a container 100.

[0014] The second electrode 20 is spaced apart from the first electrode 10 and located below the lower end (tip) of the first electrode 10. That is, the second electrode 20 is located beyond the tip of the first electrode 10, and the first electrode 10 and the second electrode 20 are spaced apart. The power supply unit 30 is connected to the first electrode 10 and the second electrode 20 by wiring 12. The container 100 is located between the first electrode 10 and the second electrode 20. In this embodiment, the container 100 is placed on the second electrode 20 and is arranged so that the first electrode 10 is spaced apart from the second electrode 20. That is, the second electrode 20 is located outside the container 100.

[0015] In this embodiment, the first electrode 10 is a high-voltage electrode. The first electrode 10 in this embodiment is a rod-shaped body extending in one direction. The first electrode 10 may have a hollow structure such as a cylindrical or polygonal tube, or a solid structure such as a columnar or polygonal pillar. In particular, if the first electrode 10 has a solid structure, durability is increased and manufacturing is easy. Examples of materials for the first electrode 10 include metals such as stainless steel, copper, and tungsten, and carbon.

[0016] The thickness (outer diameter) of the first electrode 10 is, for example, 3 to 50 mm. When the first electrode 10 is in the shape of a polygonal cylinder or a polygonal column, the outer diameter is the diameter of a circumscribed circle of the cross section of the first electrode 10.

[0017] The second electrode 20 may be any electrode that functions as a ground electrode, such as a flat electrode. Examples of materials for the second electrode 20 include metals such as stainless steel, copper, and tungsten, and carbon.

[0018] In a plan view, the area of ​​the second electrode 20 is preferably larger than the area of ​​the first electrode 10. When the area of ​​the second electrode 20 is larger, the plasma can be irradiated from the first electrode 10 to the irradiation target over a wider range.

[0019] The power supply unit 30 is only required to apply a voltage between the first electrode 10 and the second electrode 20 and generate plasma between the two electrodes. The power supply unit 30 switches between starting and stopping the supply of electricity to the electrodes. The power supply unit 30 also adjusts the voltage and frequency applied to the electrodes. An example of the power supply unit 30 is a circuit connected to an external power source and including an inverter. Another example of the power supply unit 30 is a secondary battery.

[0020] The container 100 has a container body 110, a lid 120, and a connecting portion 124. The lid 120 is configured to be detachable from the container body 110. In this embodiment, the container body 110 and the lid 120 form the container body. The container body 110 has a bottom wall portion 112 and a container side wall portion 113 that rises from the periphery of the bottom wall portion 112. The container 110 has an opening 115 at its upper end that is surrounded by the container side wall portion 113. The interior of the container 110, surrounded by the bottom wall portion 112 and the container side wall portion 113, forms a storage chamber 111. The container 110 may be cylindrical or polygonal tubular.

[0021] A pipe 180 is connected to the container side wall 113 of the container 110. The inside of the pipe 180 forms a flow path that connects the inside and outside of the container 110. That is, the flow path in the pipe 180 connects the inside and outside of the storage chamber 111. The pipe 180 may be cylindrical or polygonal tubular.

[0022] The lid 120 has a lid main body 121. The lid main body 121 has a top wall 122 and a lid side wall 123 that hangs down from the periphery of the top wall 122. The lid side wall 123 tapers downward. In this embodiment, when the lid 120 and the container 110 are combined (i.e., when the lid 120 closes the opening 115), the lid side wall 123 is located on the outer surface of the container side wall 113. That is, the lid 120 of this embodiment is an externally fitted lid. Because the lid side wall 123 tapers downward, the lid 120 fits into the container 110 when the lid 120 and the container 110 are combined. Note that the lid 120 only needs to close the opening 115 to an extent that the target cells and the like in the storage chamber 111 are not directly exposed to the irradiation environment. Therefore, a gap may be formed between the lid 120 and the container 110 .

[0023] The connecting portion 124 is provided on the lid 120. The connecting portion 124 is a cylindrical or polygonal tubular member that penetrates the top wall portion 122 in the thickness direction and hangs down toward the bottom wall portion 112. The connecting portion 124 has an insertion path 125 therein. That is, the insertion path 125 extends from above to below. The connecting portion 124 has a partition portion 126 that closes the lower end of the insertion path 125. That is, the partition portion 126 is located at the lower end of the connecting portion 124 and closes the lower end of the insertion path 125. The partition portion 126 is spaced a distance D1 from the inner bottom surface of the container 110 (the surface of the bottom wall portion 112 facing the storage chamber 111). The connecting portion 124 having the partition portion 126 can further reduce the amount of target cells and the like in the storage chamber 111 that are exposed to the environment (irradiation environment) in which the irradiation step described below is performed. Furthermore, by providing the partition wall 126, a more uniform plasma irradiation process can be achieved.

[0024] In this embodiment, the connection portion 124 is a member independent of the lid body 121. When the lid body 120 covers the opening 115, the connection portion 124 is displaced up and down relative to the lid body 121. That is, the connection portion 124 can be raised and lowered up and down while the lid body 120 is attached to the container 110. By raising and lowering the connection portion 124, the distance D1 between the lower end of the connection portion 124 and the inner bottom surface of the container 110 can be changed. For example, as shown in FIGS. 1 and 2 , when the lower end of the connection portion 124 is raised, the lower end of the connection portion 124 moves away from the inner bottom surface of the container 110. When the lower end of the connection portion 124 is lowered, as shown in FIG. 3 , the lower end of the connection portion 124 moves closer to the inner bottom surface of the container 110.

[0025] The mechanism (variable mechanism) for changing the distance D1 by raising and lowering the connection portion 124 is not particularly limited. An example of the variable mechanism will be described below with reference to FIGS. 4, 5, and 13. FIGS. 4, 5, and 13 are enlarged views of region S in FIG. 1. For example, as shown in FIG. 4, a screw thread 220 may be provided on the circumferential surface of the connection portion 124 to threadably engage with the lid main body 121. Alternatively, a screw thread may be provided on the upper outer circumferential surface of the container side wall 113 and the inner circumferential surface of the lid side wall 123 to threadably engage with the upper outer circumferential surface of the container side wall 113 and the lid side wall 123. In this case, the electrode 10 may be directly fixed to the lid 120 without providing the connection portion 124. This allows for a relatively simple variable mechanism for raising and lowering the electrode 10 together with the lid 120 relative to the container 110. Alternatively, as shown in FIG. 5 , a convex portion 222 may be provided on the peripheral surface of the connecting portion 124, and a concave portion 129 of a shape corresponding to the convex portion 222 may be provided on the lid main body 121, and these may be fitted together. In this case, by providing two or more convex portions 222, one above the other, the lower end of the connecting portion 124 can be held in any position, either with the lower end lowered or with the lower end raised. Furthermore, as shown in FIG. 13 , a concave portion 223 may be provided on the peripheral surface of the connecting portion 124, and a concave portion accommodating a rubber ring 224 may be provided on the lid main body 121, with the groove width of the concave portion 223 being smaller than the diameter of the rubber ring 224. In this case, when the electrode 10 is lowered, the rubber ring 224 fits into the concave portion 223, thereby holding the electrode 10 in a predetermined position. Furthermore, since the groove width of recess 223 is smaller than the diameter of rubber ring 224, rubber ring 224 does not fit tightly into recess 223, and connecting part 124 can be raised again after plasma irradiation. Note that connecting part 124 only needs to be airtight with lid body 121 to the extent that target cells and the like in storage chamber 111 are not directly exposed to the irradiation environment. Therefore, a gap may be formed between connecting part 124 and lid body 121.

[0026] The material of the container 110 may be a conductive material or an insulating material. However, from the viewpoint of preventing localized discharge due to high voltage, an insulating material is preferable. Examples of insulating materials include resin, ceramic, glass, etc. Examples of resins include polystyrene, polyolefin, polyester, acrylic, etc. For example, if the container 100 is to be disposable, the material of the container 110 is preferably resin. Examples of conductive materials include the same materials as those of the first electrode 10.

[0027] The material of the piping 180 is the same as the material of the container 110. The material of the piping 180 may be the same as or different from the material of the container 110. The inner diameter of the piping 180 can be determined appropriately taking into consideration the volume of the container 110, etc.

[0028] The material of the lid body 121 is the same as the material of the container 110. The material of the lid body 121 may be the same as or different from the material of the container 110. From the viewpoint of generating plasma between the electrodes, at least one of the container 110 and the lid body 121 is made of an insulating material.

[0029] The material of the connecting portion 124 may be a conductive material, an insulating material, or a composite of a conductive material and an insulating material. Specific examples of conductive and insulating materials include those similar to those exemplified above as the materials of the container 110. For example, the material of the partition 126 may be a conductive material, and the material of the other components of the connecting portion 124 may be an insulating material, or the reverse combination may also be used. Furthermore, for example, the material of the entire connecting portion 124 may be an insulating material, and the material of the lid main body 121 may be an insulating material. Alternatively, for example, the lid main body 121 and the connecting portion 124 may be integrally molded from an insulating material. When the partition 126 is made of an insulating material, the thickness of the partition 126 is preferably, for example, 0.01 to 2 mm, and more preferably 0.1 to 0.7 mm. The material of the partition 126 is preferably a dielectric (solid dielectric). Examples of dielectric materials include metal oxides, ceramics (excluding metal oxides), mica, thermoplastic resins, thermosetting resins, rubber, and thermoplastic elastomers. Metal oxides such as alumina (aluminum oxide) and ceramics are more preferred as materials for the partition wall 126 of this embodiment because they are highly durable, easy to form, and less susceptible to dielectric breakdown when a voltage is applied between the electrodes. Two or more materials may be used in combination for the partition wall 126. Furthermore, the partition wall 126 may have a single-layer structure or a multilayer structure. In the case of a multilayer structure, the materials of the layers may be the same or different.

[0030] The mechanism (variable mechanism) for raising and lowering the connection portion 124 to change the distance D1 is preferably configured to change the distance DC between the electrode 10 and the object to be treated (e.g., target cells) in the container 100 to 0.1 to 5 mm. Specifically, the distance DC is the distance between the lower end of the connection portion 124 and the surface of the object to be treated (e.g., target cells). In the present invention, when the lower end of the connection portion, the lower end of the first electrode, and / or the partition wall are provided, the distances D1 and DC are based on the lowest of the lower end of the connection portion, the lower end of the first electrode, and the lower end of the partition wall (closest to the inner bottom surface of the container or container body) when the cell processing device is in use. By keeping the distance DC within the above numerical range, appropriate plasma processing can be achieved. The cell processing device 1 that adjusts the distance DC may be configured so that the user visually checks the distance DC and manually adjusts the distance DC. However, it is more preferable that the variable mechanism be an automatically driven variable mechanism that automatically measures and adjusts the distance DC. For example, the automatic drive variable mechanism includes a drive mechanism for raising and lowering the connection part 124, a distance sensor installed at the lower end of the connection part 124, and a controller. The drive mechanism is connected to the connection part 124, the distance sensor measures the distance DC to the surface of the workpiece (e.g., target cells), and the controller is connected to the distance sensor and the drive mechanism and drives the drive mechanism to adjust the distance DC within a predetermined range. The distance sensor can be a known non-contact liquid level sensor such as an ultrasonic liquid level sensor. The controller is implemented by a computer circuit including a CPU (central processing unit) or the like. There are no particular limitations on the drive mechanism as long as it can raise and lower the connection part 124 using a motor or the like. The automatic drive variable mechanism is applicable to all embodiments described herein.

[0031] The inner diameter R111 of the container 110 is not particularly limited and is, for example, 1 to 150 mm. When the container 110 is polygonal cylindrical, the inner diameter R111 is the diameter of a circumscribing circle of the container 110 in a planar view. When the container 110 is polygonal cylindrical, the number of corners of the container 110 in a planar view is not particularly limited and is, for example, 4 to 20. Note that, in a planar view, the area of ​​the inner bottom surface of the container 110 is preferably smaller than the area of ​​the second electrode 20. When the area of ​​the inner bottom surface of the container 110 is smaller than the area of ​​the second electrode 20, plasma can be irradiated over a wider area and more uniformly. The internal height H111 of the container 110 is not particularly limited and is, for example, 5 to 50 mm.

[0032] The length L125 of the insertion path 125 in the connection portion 124 may be, for example, 1 to 50 mm, as long as it is large enough to receive the first electrode 10 and determine the position of the first electrode 10. The thickness of the insertion path 125 can be determined appropriately depending on the thickness of the first electrode 10.

[0033] <Method of Using a Cell Processing Device> A method of using a cell processing device (cell processing method) will be described with reference to Figures 1 to 3, taking as an example a case where the cell processing device is used for molecule introduction. An example of a molecule introduction method (molecule introduction method) is an introduction method having a pre-culture step, a discharge step, a contact step, and an irradiation step. More specifically, the cell processing method includes a pre-culture step of culturing a material to be processed in a culture medium within the container; a discharge step of discharging the culture medium from the container; a contact step of contacting the material to be processed with molecules within the container after the discharge step; and an irradiation step of irradiating the material to be processed with plasma after the contact step, in which the position of an electrode in the irradiation step is lower than the position of the electrode in the pre-culture step.

[0034] In the pre-culture step, target cells or the like, which are the object to be treated, are cultured. A conventionally known culture method can be used as the culture method. An example of the culture method is a method of culturing target cells or the like in the container 110 (i.e., the storage chamber 111). However, in the molecule introduction method of the present invention, pre-cultured target cells or the like may be introduced into the container 110.

[0035] The opening 115 is closed with the lid 120. At this time, the connection part 124 is raised to widen the distance D1, and the lid 120 is then attached to the container 110. Next, the flow path in the piping 180 is opened, and the target cells 44 and the culture solution 42a are injected into the container 110 ( FIG. 2 ). In the storage chamber 111, the lower end of the connection part 124 is raised, widening the distance D1, so that the culture solution 42a does not come into contact with the connection part 124. The target cells 44 and the culture solution 42a are cultured in the container 110. After the culture, the flow path in the piping 180 is opened, and the culture solution 42a is removed from the container 110, yielding the pre-cultured target cells 44 ( FIG. 1 ). In the pre-culture step, the culture solution 42 a and the target cells 44 may be introduced into the container 110 through the opening 115 of the container 110 , and then the lid 120 may be attached to the container 110 .

[0036] Examples of means for injecting the culture solution 42a etc. into the container 110 through the pipe 180 or discharging the culture solution 42a etc. from the container 110 include a pump, a syringe, and the like.

[0037] Examples of target cells etc. 44 include animal cells including human cells, non-human animal individuals, cells collected from human individuals, cells within human individuals, human individuals, plant cells, and microorganisms. Note that cells collected from human individuals include cells that are not intended to be returned to human individuals, and cells that are intended to be returned to human individuals for use in regenerative medicine, etc. Furthermore, the above-mentioned cells collected from human individuals also include cells cultured from cells collected from human individuals.

[0038] The culture medium 42a used for the pre-culture is not particularly limited and may be appropriately selected depending on the purpose. The culture conditions for the pre-culture step are, for example, 30 to 40°C and 24 to 72 hours.

[0039] The number of target cells 44 after pre-culture is determined appropriately taking into consideration the cell type, the capacity of the container 100, etc. For example, when the container 110 has a size equivalent to one well of a 96-well culture plate, the number of target cells 44 after pre-culture is 1 x 10 ３ ~1 x 10 ６For example, if the container 110 has a size equivalent to one well of a six-well container, the number of target cells 44 after pre-culture is 1×10 ４ ~1 x 10 ７ It is considered to be an individual.

[0040] The contacting step is a step in which target cells and molecules are allowed to coexist and come into contact with each other. In the contacting step, for example, with the distance D1 widened, molecules 42 are introduced from a pipe 180 into the container 110 from which the culture medium 42a has been removed, and then the flow path of the pipe 180 is closed (FIG. 3). At this time, the molecules may be made into an aqueous solution or dispersion (collectively sometimes referred to as an "aqueous liquid"), and this aqueous liquid may be injected into the container.

[0041] The molecule 42 is a molecule selected for introduction into a target cell or the like 44. Examples of the molecule 42 include a polymeric compound, a low-molecular-weight physiologically active substance, and a drug candidate. Examples of the polymeric compound include polynucleotides such as DNA and RNA or derivatives thereof, proteins or peptides such as signal transduction proteins and transcriptional regulatory factors, and derivatives thereof. As the DNA or RNA, constructs, plasmids, phagemids, and the like that are capable of expressing genes in the target cell or the like are preferred. The molecule 42 may be one type alone or a combination of two or more types.

[0042] The irradiation step is a step of irradiating plasma into the storage chamber 111 in a state where the target cells, etc. 44 and the molecules 42 coexist and are in contact with each other. Through the irradiation step, the molecules 42 are introduced into the target cells, etc. 44.

[0043] As shown in FIG. 3 , in a container 100 containing molecules 42 and target cells, etc. 44, the connector 124 is lowered to bring the lower end of the connector 124 closer to the inner bottom surface of the container 110. That is, the distance D1 between the lower end of the connector 124 and the inner bottom surface of the container 110 is narrowed, bringing the first electrode 10 closer to the target to be irradiated. Note that if the first electrode 10 is to be installed on the connector 124 when or after the container 100 is incorporated into the cell processing device 1, the connector 124 may be lowered to a predetermined position before the first electrode 10 is inserted into the insertion path 125. Next, power is supplied to the first electrode 10 from the power supply unit 30, and a voltage is applied between the first electrode 10 and the second electrode 20. When a voltage is applied between the electrodes, plasma is generated from the first electrode 10 toward the second electrode 20 (generating plasma between the electrodes by applying a voltage between the first electrode 10 and the second electrode 20 is also referred to as "plasma irradiation"). Plasma irradiation allows the molecules 42 to be introduced into the target cells, etc. 44. During plasma irradiation, the first electrode 10 approaches the irradiation target but is separated from it, so that the plasma is irradiated onto the irradiation target via the gas phase.

[0044] The distance from the lower end of the first electrode 10 to the irradiation target (a liquid in which the molecules 42 and the target cells 44 etc. coexist) is preferably 0.1 to 5 mm. When the distance from the lower end of the first electrode 10 to the irradiation target is within the above range, plasma can be generated more stably.

[0045] The AC voltage applied between the first electrode 10 and the second electrode 20 is preferably 1 to 30 kVpp, and more preferably 5 to 20 kVpp. Setting the AC voltage to the above-mentioned lower limit or higher can further increase the introduction efficiency. Setting the AC voltage to the above-mentioned upper limit or lower can suppress the temperature rise of the electrodes, thereby suppressing the temperature rise inside the container 100. The unit "Vpp (Volt peak to peak)" that represents the AC voltage is the potential difference between the maximum value and the minimum value of the AC voltage waveform.

[0046] The frequency of the alternating current applied between the first electrode 10 and the second electrode 20 is preferably 0.1 to 500 kHz, more preferably 10 to 200 kHz. Setting the frequency to the above lower limit or higher can further increase the introduction efficiency. Setting the frequency to the above upper limit or lower can further reduce damage to target cells, etc.

[0047] The plasma irradiation time in the irradiation step is, for example, preferably 10 nanoseconds to 5 seconds, more preferably 10 nanoseconds to 1 second, and more preferably 100 nanoseconds to 0.1 seconds. If the irradiation time is equal to or greater than the above-mentioned lower limit, the amount of molecules 42 introduced into target cells 44 can be further increased. If the irradiation time is equal to or less than the above-mentioned upper limit, damage to target cells can be further reduced.

[0048] In the irradiation step, the environment in the storage chamber 111 is not particularly limited, but may be, for example, CO ２ The CO in the storage chamber 111 may be present. ２ The concentration is, for example, 1 to 10% by volume.

[0049] After a given irradiation time has elapsed, the plasma irradiation is stopped, the flow path of the pipe 180 is opened, and the molecules 42 are discharged outside the vessel 110. However, it is not always necessary to discharge the molecules 42 after the plasma irradiation.

[0050] The molecule introduction method may include a post-culture step after the irradiation step. The post-culture step is a step of culturing target cells, etc. into which molecules have been introduced (sometimes referred to as "introduced cells, etc."). The culture conditions for the post-culture step are the same as those for the pre-culture step and can be determined appropriately depending on the type of introduced cells, etc., the intended use of the introduced cells, etc., etc. The culture conditions for the post-culture step may be the same as or different from those for the pre-culture step. An example of the post-culture step of this embodiment will be described. With the lid 120 attached to the container 110, the connection part 124 is raised to widen the distance D1. The culture medium 42a is injected into the container 110 through the flow path of the piping 180, the flow path of the piping 180 is closed, and the cells are cultured ( FIG. 2 ).

[0051] The transfected cells thus obtained are used, for example, for substance production, gene therapy, and the like.

[0052] According to this embodiment, some or all of the pre-incubation step, discharge step, contact step, and irradiation step can be performed with the lid 120 attached to the container 110. Therefore, target cells, etc. are not exposed to the internal space of the cell processing device 1 (e.g., the space for placing the container inside the device) or the space where the irradiation step is performed (e.g., the cell culture room, safety cabinet, clean bench). This prevents contamination by bacteria, such as target cells from other lots. Additionally, in the irradiation step, the electrodes can be brought closer to the target cells, etc., thereby improving the efficiency of introducing molecules into the target cells, etc.

[0053] Second Embodiment <Cell Processing Device> A cell processing device according to a second embodiment will be described. The configurations different from the first embodiment will be mainly described, and descriptions of other configurations will be omitted. This embodiment differs from the first embodiment in whether the lid has a conductive member or not.

[0054] The cell processing device 1a in FIG. 6 includes a container 100a having a container body 110, a lid 120a, and a connecting portion 124a. The connecting portion 124a is provided on the lid 120a. The connecting portion 124a includes a cylindrical or polygonal guide portion 128 that hangs down from the underside of the top wall portion 122 toward the bottom wall portion 112, a partition portion 126 that closes the lower end of an insertion path 125 within the guide portion 128, and a conductive member 127 located within the insertion path 125. That is, the connecting portion 124a includes the conductive member 127 that penetrates the lid body 121. Note that, although the connecting portion 124a includes the partition portion 126 in this embodiment, it may also be configured not to include the partition portion 126 (i.e., the conductive member 127 is exposed to the storage chamber 111) as long as the guide portion 128 and the conductive member 127 are hermetically joined. Furthermore, the connection portion 124a may have an irradiation portion on the lower surface of the partition portion 126 (i.e., the surface facing the second electrode 20). Examples of the irradiation portion include a flat plate with a mesh-like, circular, or polygonal hole shape (e.g., a punched metal-like), a thin film made of a conductive material patterned into a mesh-like, circular, or polygonal hole shape, and the like. In this case, the irradiation portion is connected to the conductive member 127 in the insertion path. The conductive member connected to the irradiation portion may be cylindrical, rectangular, columnar, or prismatic, or may be a conductive wire such as a copper wire, or a pattern printed on the lid 120a or the like. Alternatively, the above-mentioned irradiation portion may be provided instead of the partition portion 126.

[0055] The first electrode 10a may be any electrode as long as it can be connected to the conductive member 127, and may have a shape similar to that of the first electrode 10 in the first embodiment. Furthermore, the first electrode 10a may be, for example, a flat electrode. The material of the first electrode 10a is the same as that of the first electrode 10.

[0056] The material of the guide portion 128 may be a conductive material or an insulating material. For example, the guide portion 128 may be made of the same material as the lid main body 121 and may be molded integrally with the lid main body 121. The material of the partition portion 126 is the same as the material of the guide portion 128. The material of the partition portion 126 may be the same as or different from the material of the guide portion 128. Note that when the connecting portion 124a has an irradiation portion on the lower surface of the partition portion 126 and the partition portion 126 is made of a conductive material, the conductive member 127 is connected to the irradiation portion via the partition portion 126 made of a conductive material. When the connecting portion 124a has an irradiation portion on the lower surface of the partition portion 126 and the partition portion 126 is made of an insulating material, the partition portion 126 has another conductive member that connects the conductive member 127 and the irradiation portion. In other words, the irradiation portion is electrically connected to the conductive member 127.

[0057] The connecting portion 124a is similar to the connecting portion 124 in the first embodiment. That is, the connecting portion 124a is a member independent of the lid body 121, and can be raised and lowered up and down in a state in which the lid body 120 is attached to the container 110. The variable mechanism in this embodiment is similar to the variable mechanism in the first embodiment.

[0058] The material of the conductive member 127 is the same as the material of the first electrode 10a. The material of the conductive member 127 and the material of the first electrode 10a may be the same or different.

[0059] <Method of Using the Cell Processing Apparatus> An example of a method of using the cell processing apparatus (cell processing method) of this embodiment will be described using a molecule introduction method as an example. An example of the molecule introduction method is an introduction method having a pre-incubation step, a discharge step, a contact step, and an irradiation step. The pre-incubation step, the discharge step, and the contact step are the same as in the first embodiment.

[0060] The irradiation step of this embodiment is the same as that of the first embodiment, except that the first electrode 10a is connected to the conductive member 127 of the container 100a.

[0061] In addition, the molecule introduction method of this embodiment may include the post-culture step described above in the first embodiment.

[0062] (Third embodiment) <Cell processing device> A cell processing device according to a third embodiment will be described. The configuration that differs from the first and second embodiments will be mainly described, and a description of other configurations will be omitted. This embodiment differs from the first and second embodiments in that the lid and the container are integrated. An example of the container 100b of this embodiment is a so-called microchannel device.

[0063] The cell processing device 1b in Fig. 7 has a container 100b. The container 100b has a container body 110b, a lid body 120b, and a connecting portion 124, and the container body 110b and the lid body 120b are inseparably integrated. "Inseparably integrated" means that they are joined to such an extent that they cannot be easily separated.

[0064] The container 110b has a bottom wall 112 and a container side wall 113b rising from the periphery of the bottom wall 112. The lid 120b has a flat lid body 121b. A connecting portion 124 is provided on the lid 120b. Note that this embodiment may have a connecting portion 124a (second embodiment) instead of the connecting portion 124. The variable mechanism in this embodiment is similar to the variable mechanism in the first embodiment. The lid 120b closes the opening 115 of the container 110b. Note that, because the container is highly airtight in this embodiment, a flow path serving as an air vent may be provided in the upper part of the container (e.g., the lid 120b) to smoothly inject and discharge the culture medium and the like into and from the container via the flow path in the piping 180. It is preferable to provide a sterilizing filter in such a vent.

[0065] <Method of Using Cell Processing Device> An example of a method of using the cell processing device (cell processing method) of this embodiment will be described using a molecule introduction method as an example. The molecule introduction method includes a cell filling step, a pre-incubation step, a discharge step, a contact step, and an irradiation step. The irradiation step is the same as in the first embodiment.

[0066] The cell filling process in this embodiment will be described. With the opening 115 closed by the lid 120b, the connector 124 is raised to widen the distance D1. Next, the flow path in the piping 180 is opened, and the target cells 44 and the culture medium 42a are injected into the vessel 110b (FIG. 7).

[0067] After the cell filling step, the target cells 44 in the storage chamber 111 are cultured (pre-culture step) in the same manner as in the first embodiment.

[0068] After the pre-culture step, the flow path of the piping 180 is opened and the culture solution is discharged. Next, the molecules 42 are injected into the piping 180. The injected molecules 42 are filled into the storage chamber 111 (for example, a well in a microchannel device) via the piping 180. In this way, the molecules 42 are filled into the container 110b, and the molecules 42 are brought into contact with the target cells, etc., in the storage chamber 111 (contact step). Next, with the molecules 42 and the target cells, etc., 44 in contact with each other, an irradiation step is performed, thereby introducing the molecules 42 into the target cells, etc., 44.

[0069] In addition, the molecule introduction method of this embodiment may include a post-culture step.

[0070] According to this embodiment, the container 110b and the lid 120b are integrated to create a cleaner environment inside the storage chamber 111, thereby more effectively preventing contamination.

[0071] (Fourth embodiment) <Cell processing device> A cell processing device according to the fourth embodiment will be described. The configuration that differs from the first to third embodiments will be mainly described, and a description of other configurations will be omitted. This embodiment differs from the first to third embodiments in that the container does not have a lid and that an air vent is provided at the top of the container.

[0072] The cell processing device 1d of FIG. 9 includes a container 100d, a first electrode 10d, a connecting portion 124d (variable mechanism), and a conductive member 127. The container 100d has a bottom wall 112d, a container side wall 113d rising from the periphery of the bottom wall 112d, and a top wall 117d provided at the upper end of the container side wall 113d. The interior of the container 100d, surrounded by the bottom wall 112d and the container side wall 113d, forms a storage chamber 111d. The top wall 117d is provided with an air vent 180a. The presence of this air vent 180a allows for smooth injection and discharge of culture medium and other fluids into and from the container via the flow path in the piping 180. A sterilizing filter is preferably provided within the air vent 180a. The bottom wall 112d, the container side wall 113d, and the top wall 117d are preferably integrally formed. The container 100d may be cylindrical or polygonal tubular. In the cell processing device 1d of this embodiment, the lid does not need to be attached or detached, so contamination can be prevented more reliably.

[0073] The second embodiment is similar to the second embodiment except that the connecting portion 124d is provided on the top wall portion 117d rather than on the lid, and no partition is provided. The configuration of the first electrode 10d is also similar to the second embodiment, and the configuration of the conductive member 127 is also similar to the second embodiment. When the variable mechanism of the connecting portion 124d is the above-mentioned automatically driven variable mechanism, the distance DC in this embodiment is the distance between the lower end of the electrode 10d and the surface of the object to be treated (e.g., target cells).

[0074] <Method of Using the Cell Processing Device> An example of a method of using the cell processing device (cell processing method) of this embodiment will be described using a molecule introduction method as an example. When carrying out this method, the same power supply unit 30 and second electrode 20 as in the first embodiment are arranged. An example of the molecule introduction method is an introduction method having a pre-incubation step, a discharge step, a contact step, and an irradiation step. The details of the pre-incubation step, the discharge step, the contact step, and the irradiation step can be applied mutatis mutandis to the explanations in the first embodiment.

[0075] In addition, the molecule introduction method of this embodiment may include the post-culture step described above in the first embodiment.

[0076] (Fifth Embodiment) <Cell Processing Device> A cell processing device according to the fifth embodiment will be described. The configuration that differs from the first to fourth embodiments will be mainly described, and a description of other configurations will be omitted. This embodiment is similar to the first and second embodiments, except that the container body is not divided into a container body and a lid body.

[0077] The cell processing device 1e of FIG. 10 includes a container 100e, a first electrode 10e, and a connecting portion 124e (adjustable mechanism). A partition wall 126e is provided at the lower end of the first electrode 10e. The container 110e includes a bottom wall 112e, a container side wall 113e rising from the periphery of the bottom wall 112e, and a top wall 117e provided at the upper end of the container side wall 113e. In the container 110e, the interior surrounded by the bottom wall 112e and the container side wall 113e forms a storage chamber 111e. The bottom wall 112e, the container side wall 113e, and the top wall 117e are preferably integrally formed. The container 110e may be cylindrical or polygonal. The cell processing device 1e of this embodiment does not require the lid to be attached or detached, thereby more reliably preventing contamination.

[0078] The connecting portion 124e is the same as that in the first and second embodiments, except that it is provided on the top wall portion 117e rather than on the lid. The partition portion 126e is preferably made of a dielectric material such as the ceramic described above.

[0079] <Method of Using the Cell Processing Device> An example of a method of using the cell processing device of this embodiment (cell processing method) will be described using a molecule introduction method as an example. When carrying out this method, the same power supply unit 30 and second electrode 20 as in the first embodiment are arranged. An example of a molecule introduction method is an introduction method having a pre-incubation step, a discharge step, a contact step, and an irradiation step. The details of the pre-incubation step, the discharge step, the contact step, and the irradiation step can be applied mutatis mutandis to the explanations in the first and second embodiments. That is, the container of this embodiment is structurally different from the container of the first and second embodiments mainly in that it is not separated into a container body and a lid body. However, taking such differences into consideration, the explanations in the first and second embodiments can be applied to this embodiment with appropriate modifications.

[0080] In addition, the molecule introduction method of this embodiment may include the post-culture step described above in the first embodiment.

[0081] (Sixth Embodiment) <Cell Processing Device> A cell processing device according to the sixth embodiment will be described. The configuration that differs from the first to fifth embodiments will be mainly described, and a description of other configurations will be omitted. This embodiment is similar to the fifth embodiment except that the electrode 10e is not provided, the connection portion is made of a conductive material and functions as the first electrode, and the partition portion is made of a dielectric material. In other words, in this embodiment, the first electrode has the same structure and function as the connection portion 124e of the fifth embodiment.

[0082] The cell processing device 1f of FIG. 11 includes a container 100f and a first electrode 124f. A partition wall 126f is provided at the lower end of the first electrode 124f. The container 100f has a bottom wall 112f, a container side wall 113f rising from the periphery of the bottom wall 112f, and a top wall 117f provided at the upper end of the container side wall 113f. In the container 100f, the interior surrounded by the bottom wall 112f and the container side wall 113f forms a storage chamber 111f. The bottom wall 112f, the container side wall 113f, and the top wall 117f are preferably integrally formed. The container 100f may be cylindrical or polygonal. The cell processing device 1f of this embodiment does not require the lid to be attached or detached, thereby more reliably preventing contamination.

[0083] The electrode 124f of this embodiment has the same shape as the connection part 124 of the first embodiment, and is preferably made of a dielectric material such as ceramic, glass, or silicone-based resin, and more preferably ceramic. As in the first embodiment, the container 100f of this embodiment also has a mechanism (variable mechanism) that raises and lowers the connection part 124f to change the distance D1. In addition, the partition part 126f is also preferably made of a dielectric material such as ceramic.

[0084] <Method of Using the Cell Processing Device> An example of a method of using the cell processing device of this embodiment (cell processing method) will be described using a molecule introduction method as an example. When carrying out this method, the same power supply unit 30 and second electrode 20 as in the first embodiment are arranged. An example of a molecule introduction method is an introduction method having a pre-incubation step, a discharge step, a contact step, and an irradiation step. The details of the pre-incubation step, the discharge step, the contact step, and the irradiation step can be applied mutatis mutandis to the explanations in the first and second embodiments. That is, the container of this embodiment is structurally different from the container of the first and second embodiments mainly in that it is not separated into a container body and a lid body. However, taking such differences into consideration, the explanations in the first and second embodiments can be applied to this embodiment with appropriate modifications.

[0085] In addition, the molecule introduction method of this embodiment may include the post-culture step described above in the first embodiment.

[0086] Seventh Embodiment <Cell Processing Apparatus> A cell processing apparatus according to the seventh embodiment will be described with reference to FIG. 12 . The following mainly describes the configurations that differ from the first to sixth embodiments, and a description of other configurations will be omitted. This embodiment is similar to the fifth embodiment except for the following: the container 100g is divided into a container lower portion 100g1 and a container upper portion 100g2; threads 220 are provided on the outer surfaces of the side walls of the container lower portion 100g1 and the container upper portion 100g2, which are configured to threadably engage with the nut-shaped variable mechanism 124g; and the first electrode 10g is fixed directly to the top wall portion 117d of the container upper portion 100g2 without a connecting portion. In this embodiment, the container lower portion 100g1 and the container upper portion 100g2 are threadably engaged with the nut-shaped variable mechanism 124g as a bolt. For example, when the lower container portion 100g1, the upper container portion 100g2, and the nut-shaped variable mechanism 124g are combined, a gap is left between the upper end of the lower container portion 100g1 and the lower end of the upper container portion 100g2, and only one of the lower container portion 100g1, the upper container portion 100g2, and the nut-shaped variable mechanism 124g is rotated in the screw direction to adjust the distance D1 (however, in this embodiment, the distance between the lower end of the electrode 10g and the inner bottom surface of the lower container portion 100g1) or the distance DC (however, in this embodiment, the distance between the lower end of the electrode 10g and the surface of the workpiece in the container 100g). Furthermore, in this embodiment, the variable mechanism 124g is provided on the side of the container 100g, thereby realizing a variable mechanism with a relatively simple structure.

[0087] The material of the nut-shaped variable mechanism 124g is not particularly limited, and examples thereof include the same materials as those listed as the materials of the connecting portion 124 in the first embodiment. The specific shape and dimensions of the nut-shaped variable mechanism 124g are not particularly limited as long as the above-mentioned distance D1 and distance DC can be easily adjusted, but from the viewpoint of facilitating rotation or fixation of the nut-shaped variable mechanism 124g, a shape such as a hexagonal nut or a wing nut is preferable.

[0088] <Method of Using the Cell Processing Device> An example of a method of using the cell processing device of this embodiment (cell processing method) will be described using a molecule introduction method as an example. When carrying out this method, the same power supply unit 30 and second electrode 20 as in the first embodiment are arranged. An example of a molecule introduction method is an introduction method having a pre-incubation step, a discharge step, a contact step, and an irradiation step. The details of the pre-incubation step, the discharge step, the contact step, and the irradiation step can be applied mutatis mutandis to the explanations in the first and second embodiments. That is, the container of this embodiment is structurally different from the container of the first and second embodiments mainly in that it is not separated into a container body and a lid body. However, taking such differences into consideration, the explanations in the first and second embodiments can be applied to this embodiment with appropriate modifications.

[0089] In addition, the molecule introduction method of this embodiment may include the post-culture step described above in the first embodiment.

[0090] 1 to 3, 6, 7, 9, and 10 show the state in which the first electrode is installed in the container of the present invention, but the first electrode may be incorporated into the container in advance or may be incorporated into the container later when the cell processing device is used. In the case where the first electrode is incorporated into the container later when the cell processing device is used, it is preferable that the container has a partition wall.

[0091] (Uses) The plasma cell treatment device of the present invention can stimulate target cells, etc., which are the treatment objects, while mitigating damage to them. Therefore, the plasma cell treatment device of the present invention is suitable for cell treatment such as cell proliferation and molecule introduction. In particular, the plasma cell treatment device of the present invention is more suitable for molecule introduction.

[0092] (Other Embodiments) In the first to seventh embodiments, the container has piping, but the present invention is not limited thereto. For example, the container does not need to be connected to piping. However, from the viewpoint of performing some or all of the pre-incubation step, discharge step, contact step, irradiation step, and post-incubation step with the lid attached to the container, it is preferable that the container has piping.

[0093] In the second embodiment, the conductive member is rod-shaped, but the present invention is not limited to this. When the cover has a conductive member, the conductive member may be plate-shaped.

[0094] In the first and second embodiments, the lid body is an outer fitting lid, but the lid body of the present invention may be a so-called inner fitting lid.

[0095] In the first to fifth embodiments, the connection portion has an insertion path, but the present invention is not limited to this. For example, the connection portion may be a metal plate, metal foil, or the like located on the upper surface of the lid body. However, from the viewpoint of optimizing the discharge distance, it is preferable to provide an insertion path to bring the first electrode or the conductive member closer to the irradiation target.

[0096] In the first to seventh embodiments, the container is placed on the second electrode (i.e., the second electrode is located on the lower surface of the bottom wall of the container), but the present invention is not limited to this. For example, as shown in FIG. 8, the second electrode 20c may be an electrode that runs around the container side wall 113 near the bottom wall 112. When the second electrode 20c is used, it is preferable that the lower end 21 of the second electrode 20c is located below the upper surface 41 of the irradiation target. The container may also have a conductive member corresponding to the second electrode.

[0097] In the first to third embodiments, the container is a combination of one container having one storage chamber and one lid, but the present invention is not limited to this. For example, the container of the present invention may be a combination of one container having two or more storage chambers and one lid that closes the opening of the container. Alternatively, the container of the present invention may be a combination of one container having two or more storage chambers and two or more lids that close each of the storage chambers.

[0098] In the first to seventh embodiments, one connection part is provided for one container or vessel, but two or more connection parts may be provided for one container or vessel so that two or more first electrodes act on one container or vessel.

[0099] In the first to seventh embodiments, one pipe is provided for one vessel or container, but the present invention is not limited thereto, and two or more pipes may be provided for one vessel or container. By providing two or more pipes, it is possible to separate the injection and waste liquid paths, or to separate the injection of cells and culture medium from the injection of DNA. Furthermore, a check valve may be provided in the pipe. A known check valve can be used. For example, a check valve such as that shown in Figures 1 to 8 of WO 2012 / 147463 can be used.

[0100] Alternatively, molecules may be introduced by surface discharge. Instead of arranging the first and second electrodes through the bottom wall of the container body, multiple electrodes may be arranged at the top of the container body, and a voltage may be applied between the multiple electrodes to generate a discharge between the multiple electrodes and the irradiation target where target cells and molecules coexist. In this case, multiple needle-shaped electrodes are typically used. For specific electrode configurations in the case of surface discharge, see, for example, Republished 2019 / 244895. The connection portion may be provided for each of the multiple electrodes, or the multiple electrodes may be housed together in one or more connection portions.

[0101] 1, 1a, 1b, 1d, 1e, 1f, 1g Cell processing device 10, 10a, 10d, 10e, 124f, 10g First electrode 20, 20c Second electrode 42 Molecule 44 Target cell or the like 100, 100a, 100b, 100d, 100e, 100f, 100g Container 111, 111d, 111e, 111f, 111g Storage chamber 110, 110b Receptacle 115 Opening 120, 120a, 120b Lid 121, 121b Lid main body 124, 124a, 124b, 124d, 124e Connection part 125 Insertion path 126, 126e, 126f Partition part 127 Conductive member 180 Piping 220 Thread 222 Convex part

Claims

1. A container for a plasma cell processing device that processes an object to be processed with plasma, comprising: a container body; an electrode installed in the upper part of the container body to generate plasma; a processing section installed in the lower part of the container body; a flow path connecting the inside and outside of the container body; and a variable mechanism that changes the relative position between the electrode and the processing section.

2. A container for a plasma cell processing device as described in claim 1, wherein the variable mechanism has a connection part that connects the container body and the electrode.

3. A container for a plasma cell processing device as described in claim 1, wherein the variable mechanism is provided on the side of the container body.

4. A container for a plasma cell processing device as described in claim 1 or 2, wherein the variable mechanism is configured to change the distance between the electrode and the processing section within the container body to 0.1 to 5 mm.

5. A container for a plasma cell processing device according to claim 1 or 2, wherein the container body is composed of a plurality of parts that are detachable from one another.

6. A container for a plasma cell treatment device according to claim 1 or 2, wherein the electrode is a needle electrode.

7. A container for a plasma cell processing device according to claim 1 or 2, wherein the electrode has a partition wall made of a dielectric material at the lower end.

8. A cell processing method using a plasma cell processing device equipped with a container as defined in claim 1 or 2, comprising: a pre-culture step of culturing an object to be processed in a culture medium within the container; a discharge step of discharging the culture medium from the container; a contact step of bringing molecules into contact with the object to be processed within the container after the discharge step; and an irradiation step of irradiating the object to be processed with plasma after the contact step, wherein the position of the electrode in the irradiation step is lower than the position of the electrode in the pre-culture step.

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