JP2023154331A - Sample discharge system, sample discharge device, sample discharge method, and method for producing artificial tissue - Google Patents

Abstract

To provide a sample discharge system, a sample discharge device, a sample discharge method with an easy discharge operation of a sample, and a method for producing an artificial tissue.SOLUTION: In a sample discharge system comprising a sample discharge assembly for discharging a sample, the sample discharge assembly comprises: a discharge part having a sample space and a discharge port for discharging a sample housed in the sample space to the outside; a gas housing part having a gas chamber communicating with the sample space, and capable of housing gas in the gas chamber; a pressurization mechanism for increasing the pressure inside the gas chamber; and a valve mechanism having a valve chamber communicating with the gas chamber and a valve body capable of switching a closed configuration and an open configuration, where the valve chamber communicates with a vent part open to an outer space, the valve body blocks communication between the valve chamber and the vent part in the closed configuration, and allows the valve chamber and the vent part to communicate with each other in the open configuration.SELECTED DRAWING: Figure 1

Description

Embodiments of the present invention relate to a sample discharging system, a sample discharging device, a sample discharging method, and a method for manufacturing an artificial tissue.

2. Description of the Related Art A sample ejecting device for ejecting a fluid sample and injecting it into a target is known. Patent Document 1 describes a syringe pump that can inject a medicinal solution inhaled into a syringe into a living body at a predetermined rate.

Japanese Patent Application Publication No. 6-296690

However, in such a sample discharging device, the operability of the discharging operation may not be sufficient depending on the properties of the sample and the type of target into which the sample is injected.

The problem to be solved by the present invention is to provide a sample discharging system, a sample discharging device, a sample discharging method, and a method for manufacturing an artificial tissue that allow easy sample discharging operation.

The present invention may include the following aspects.
[1] A sample discharge system including a sample discharge assembly configured to discharge a sample, the sample discharge assembly including a sample space and a discharge section having a discharge port configured to discharge the sample accommodated in the sample space to the outside; a gas storage section having a gas chamber communicating with the sample space and capable of containing gas in the gas chamber; a pressurizing mechanism for increasing the pressure within the gas chamber; a valve chamber communicating with the gas chamber; and a closed configuration. and a valve mechanism having a valve body capable of switching between the valve chamber and the open configuration, the valve chamber communicating with a vent portion open to an external space, and the valve body communicating with the valve chamber in the closed configuration. A sample discharge system that blocks communication with a vent section and allows communication between the valve chamber and the vent section in the open configuration.
[2] The sample discharge system according to [1], wherein the pressurizing mechanism compresses the gas in the gas chamber to push out the sample from the discharge port via the gas.
[3] When the valve body is in the closed configuration, an internal space communicating with the discharge port is formed inside the sample discharge assembly, and the internal space, except for a portion connected to the discharge port, The sample discharge system according to [1] or [2], which is closed from an external space.
[4] The sample discharge system according to [3], wherein the pressurizing mechanism changes the volume of the internal space.
[5] The valve mechanism includes a solenoid and a plunger that moves under the action of the solenoid, and the plunger moves the valve body between the closed configuration and the open configuration as the plunger moves under the action of the solenoid. The sample discharge system according to [1] or [2], which switches between.
[6] The valve body switches between the closed configuration and the open configuration by moving, and when the solenoid acts on the plunger, the plunger switches the valve body from the closed configuration to the open configuration. The sample discharge system according to [5], which moves the sample.
[7] The valve body switches between the closed configuration and the open configuration by moving, and the valve mechanism directly or indirectly attaches the valve body in a direction from the open configuration to the closed configuration. The sample ejection system according to [5], comprising an elastic member that exerts force.
[8] The pressurizing mechanism includes a piston body that is at least partially inserted into the gas chamber, and a piston drive mechanism that moves the piston body within the gas chamber, [1] or [2] The sample ejection system described in .
[9] The sample discharge system according to [1] or [2], further comprising a control unit that electrically controls operation of at least one of the pressurizing mechanism and the valve mechanism.
[10] A sample discharging device for discharging a sample from a discharging section having a sample space capable of accommodating a sample therein, the gas containing section being coupled to the discharging section, wherein the gas containing section is connected to the discharging section. a gas storage section having a gas chamber communicating with the sample space when coupled to the discharge section; a pressurizing mechanism for increasing the pressure within the gas chamber; a valve chamber communicating with the gas chamber; a valve mechanism having a valve body capable of switching the configuration, the valve chamber communicates with a vent portion open to external space, and the valve body communicates with the valve chamber and the vent portion in the closed configuration. A sample discharging device, wherein the valve chamber and the vent portion communicate with each other in the open configuration.
[11] A method for discharging a sample from a discharging section having a sample space inside which can accommodate a sample, comprising the step of accommodating a sample in the sample space, and increasing the internal pressure of the sample space. pushing out a sample from the discharge portion; and switching a valve body capable of switching between a closed configuration and an open configuration from the closed configuration to an open configuration, wherein the valve body is configured to be connected to the sample space in the closed configuration. A method for discharging a sample, comprising the steps of: blocking communication with an external space, and allowing communication between the sample space and the external space in the open configuration.
[12] Further comprising the step of inserting the discharge section containing the sample into a target (excluding a human), and in the step of pushing out the sample from the discharge section, the sample is injected into the target; [ 11].
[13] The sample discharging method according to [12], wherein the target is a fetus of a non-human animal.
[14] The sample discharging method according to any one of [11] to [13], wherein the sample contains cells.
[15] A method for producing an artificial tissue, which includes the steps of: inserting a discharge part containing cells in an internal sample space into a fetus of a non-human animal; and increasing the pressure of gas in the sample space to remove the cells. injecting the cells into the fetus by extruding the cells from the discharge portion; and switching a valve body capable of switching between a closed configuration and an open configuration from the closed configuration to the open configuration, the valve body The method further comprises: blocking communication between the sample space and external space in the closed configuration and allowing communication between the sample space and external space in the open configuration; and pulling out the discharge part from the fetus. A method for producing an artificial tissue, comprising the step of growing the injected cells in the body of the fetus.

According to the present invention, it is possible to provide a sample discharging system, a sample discharging apparatus, a sample discharging method, and a method for manufacturing an artificial tissue that allow easy sample discharging operation.

1 is a schematic diagram showing the overall configuration of a sample discharge system according to an embodiment. FIG. 2 is a perspective view of a sample ejection assembly according to an embodiment. FIG. 3 is a cross-sectional view taken along line III-III of FIG. 2 illustrating a sample ejector assembly according to an embodiment. 4 is a cross-sectional view taken along line IV-IV of FIG. 3 illustrating a sample delivery assembly according to an embodiment; FIG. FIG. 4 is a cross-sectional view taken along line V-V of FIG. 3 illustrating a sample delivery assembly according to an embodiment. FIG. 4 is an enlarged cross-sectional view of a portion (area E in FIG. 3) of the sample ejection assembly (closed state) according to the embodiment. FIG. 4 is an enlarged cross-sectional view of a portion (area E in FIG. 3) of the sample ejection assembly (in an open state) according to an embodiment. FIG. 1 is a block diagram showing the configuration of a sample discharge system according to an embodiment. 3 is a flowchart showing the operation of the sample discharge system according to the embodiment. FIG. 2 is a photograph of the kidney development area of a mouse into which kidney progenitor cells were transplanted using a sample discharge system.

Hereinafter, a sample discharging system, a sample discharging device, a sample discharging method, and a method for manufacturing an artificial tissue according to embodiments will be described with reference to the drawings. Note that the drawings are schematic or conceptual, and the relationship between the thickness and width of each part, the size ratio between parts, etc. are not necessarily the same as those in reality. Furthermore, even when the same part is shown, the dimensions and ratios may be shown differently depending on the drawing. Furthermore, the XYZ coordinates shown in the drawings are defined for convenience of explanation and do not limit the invention.

As used herein, "based on XX" means "based on at least XX" and includes cases where it is based on another element in addition to XX. Furthermore, "based on XX" is not limited to the case where XX is used directly, but also includes the case where it is based on calculations and processing performed on XX. "XX" is an arbitrary element (for example, information).

<Configuration of sample discharge system 1>
A sample discharge system 1 according to an embodiment will be described with reference to FIGS. 1 to 8.
FIG. 1 is a schematic diagram showing the overall configuration of a sample discharge system according to an embodiment.
FIG. 2 is a perspective view of a sample ejection assembly 100 according to an embodiment.
FIG. 3 is a cross-sectional view taken along line III--III in FIG. 2, illustrating the sample ejection assembly 100 according to an embodiment.
FIG. 4 is an exploded view taken along line IV-IV in FIG. 3 of the sample ejector assembly 100 according to an embodiment.
FIG. 5 is an exploded view taken along line VV in FIG. 3 of the sample ejector assembly 100 according to an embodiment.
FIG. 6 is an enlarged view of a portion (region E in FIG. 3) of the sample ejection assembly 100 (closed state) according to an embodiment.
FIG. 7 is an enlarged view of a portion (area E in FIG. 3) of the sample ejection assembly 100 (in an open state) according to an embodiment.
FIG. 8 is a block diagram showing the configuration of the sample discharge system 1 according to the embodiment.

The sample discharge system 1 is a system for discharging a sample S. Specifically, as shown in FIG. 1, the sample discharge system 1 includes a sample discharge assembly 100, a positioning unit 200, and a control unit 300. The sample ejection assembly 100 is fixed to a positioning unit 200 and positioned with respect to the target T. Control unit 300 controls the operation of sample dispensing assembly 100. Note that in this specification, the term "system" is not limited to a system consisting of two or more devices, but also includes a system consisting of a single device.

Sample S has fluidity and viscosity. Preferably, the sample S has such fluidity that it does not substantially flow out naturally from the mouth of the capillary for at least several seconds to several tens of seconds when the capillary containing the sample S is placed horizontally. It has viscosity. The type of sample S is not particularly limited, but includes, for example, a biological sample. Preferably, sample S contains cells. Cells are not particularly limited, and may include stem cells, progenitor cells, terminally differentiated cells, and the like. Examples of cells include, but are not limited to, pluripotent stem cells such as ES cells and iPS cells, cells induced to differentiate from somatic stem cells, and cells obtained by treating biological tissue with degrading enzymes. . The cells may be genetically modified cells or non-genetically modified cells.

The target T is the target for injection of the sample S. When the sample S is a cell, the target T may be a target for transplantation of the sample S. For example, the target T is a human or non-human animal, its fetus, or all or part of a body tissue or organ thereof. The target T is not limited to an area inside a living body, but may be a body tissue or organ placed outside a living body. For example, the target T is a fetus of a non-human animal or a part thereof.

<Configuration of sample discharge assembly 100>
The sample discharge assembly 100 has a configuration for discharging a sample S. Specifically, as shown in FIG. 2, sample delivery assembly 100 includes a delivery tube 102, a syringe 104, a piston 106, and a valve mechanism 108.

[Configuration of discharge pipe 102]
As shown in FIG. 3, the discharge pipe 102 (an example of a "discharge section") accommodates the sample S therein and discharges the sample S contained therein. Specifically, discharge tube 102 is a hollow, elongated tubular member. A sample space 102A is formed inside the discharge tube 102. A discharge port 102B for discharging the sample S is formed at one end of the discharge tube 102. More specifically, the discharge tube 102 has two openings that communicate with the sample space 102A, one of the two openings is coupled to the syringe 104 and communicates with the gas chamber 22 of the syringe 104; The other one functions as the discharge port 102B.

Discharge tube 102 is removably attached to syringe 104. However, the configuration of the discharge pipe 102 is not limited to this, and the discharge pipe 102 may be attached to the syringe 104 in a non-removable manner, or may be integrally provided as a part of the syringe 104.

The discharge tube 102 may be provided with a marker such as a scale that can be used to estimate the amount of the sample S accommodated in the sample space 102A. When the user discharges the sample S from the discharge tube 102, the user can estimate the discharge amount of the sample S by visually recognizing this marker.

The material of the discharge tube 102 is not particularly limited, but is preferably a material that is not harmful to biological materials. Examples of the material for the discharge pipe 102 include glass, polymer materials, metal materials, and the like.

The discharge tube 102 can be inserted into the target T and used to inject the sample S into the target T. Preferably, the discharge tube 102 has enough strength to be able to puncture living tissue.

[Configuration of syringe 104]
The syringe 104 (an example of a "gas storage section") is coupled to the discharge tube 102 to hold the discharge tube 102, and operates to push the sample S inside the discharge tube 102 to the outside. For example, syringe 104 functions as a body that supports the entire sample dispensing assembly 100. Specifically, syringe 104 has a syringe body 10 and a manifold 12. Manifold 12 is interposed between discharge pipe 102 and syringe body 10. A first connecting portion 14 is provided between the discharge pipe 102 and the manifold 12 . The first connecting portion 14 connects the discharge pipe 102 and the manifold 12 so that they communicate with each other. A second connecting portion 16 is provided between the syringe body 10 and the manifold 12. The second connecting portion 16 connects the syringe body 10 and the manifold 12 so that they communicate with each other.

The syringe body 10 accommodates therein a gas (for example, air) for pushing out the sample S in the discharge tube 102, and discharges the sample S into the discharge tube 102 via the gas. The syringe body 10 has a first chamber 22A as an internal space. The syringe body 10 can accommodate gas in the first chamber 22A. Specifically, the syringe main body 10 is a cylindrical member with both ends open. For example, the syringe body 10 has a general syringe configuration. The material of the syringe body 10 is not particularly limited, but it is preferably made of a material that is at least partially transparent or translucent. Examples of the material of the syringe body 10 include glass, polymer materials, and the like.

A first holding part 10A and a second holding part 10B are provided on both sides of the syringe body 10. The first holding part 10A holds one end of the syringe main body 10, and the second holding part 10B holds the other end of the syringe main body 10.

The manifold 12 connects the discharge pipe 102 and the syringe body 10 in a state where they are in communication with each other. Manifold 12 forms a flow path structure within sample dispensing assembly 100 . Specifically, the manifold 12 has a second chamber 22B as an internal space that communicates the sample space 102A of the discharge tube 102 with the space inside the syringe body 10. Manifold 12 can contain gas (eg, air) in second chamber 22B. The first chamber 22A and the second chamber 22B communicate with each other to form a gas chamber 22 that can contain gas. Thus, syringe 104 can contain gas in gas chamber 22 . Gas chamber 22 communicates with sample space 102A of discharge tube 102 when syringe 104 is coupled to discharge tube 102 .

As shown in FIGS. 2 and 4, the manifold 12 has vent holes 20 (an example of a "vent part") on both side surfaces. The vent hole 20 can relieve the pressure inside the sample discharge assembly 100 by discharging the gas inside the sample discharge assembly 100 to the outside.

The second chamber 22B of the manifold 12 can form a branched flow path. Specifically, as shown in FIG. 3, the manifold 12 has a first passage 24, a second passage 26, and a third passage 28 therein. The first passageway 24 extends through the front wall of the manifold 12 from the second chamber 22B to the sample space 102A of the discharge tube 102. The second passageway 26 extends through the top wall of the manifold 12 from the second chamber 22B to the top surface 12A of the manifold 12. As shown in FIG. 4, the third passage 28 branches from the upper surface 12A of the manifold 12 inside the manifold 12 and extends to the two vent holes 20 on both sides of the manifold 12. The second passage 26 and the third passage 28 are connected by a valve mechanism 108, which will be described later.

In this way, the manifold 12 can connect the space inside the syringe body 10, the sample space 102A of the discharge tube 102, and the exhaust vent hole 20 to each other through the internal branch passage.

[Configuration of piston 106]
Piston 106 (an example of a "pressurization mechanism") can increase the pressure within gas chamber 22. The piston 106 compresses the gas in the gas chamber 22 of the syringe 104 and pushes out the sample S from the discharge port 102B of the discharge pipe 102 via this gas. Specifically, as shown in FIG. 3, the piston 106 includes a piston body 30, a piston drive mechanism 32, and a piston housing 34 that houses the piston body 30 and the piston drive mechanism 32 inside.

Piston body 30 is at least partially inserted into first chamber 22A. By sliding within the syringe body 10, the piston body 30 can change the internal volume of the sample dispensing assembly 100, thereby changing the pressure of the gas within the sample dispensing assembly 100. Specifically, as shown in FIG. 3, the piston main body 30 has a piston head 30A and a piston rod 30B.

The piston head 30A is a disc-shaped member that constitutes the distal end of the piston body 30. The outer diameter of the piston head 30A is approximately the same as the inner diameter of the syringe body 10. The piston head 30A thereby partially defines a space formed within the sample ejection assembly 100. The piston rod 30B is a rod-shaped member extending in the proximal direction (-Z direction) from the piston head 30A.

Piston drive mechanism 32 moves piston body 30 within gas chamber 22 . Specifically, the piston drive mechanism 32 drives the piston body 30 to slide within the first chamber 22A of the syringe body 10. Any known translation mechanism can be used as the piston drive mechanism 32. For example, as shown in FIG. 3, the piston drive mechanism 32 includes a motor 40, a gear 42, a feed screw 44, and a carriage 46.

The motor 40 functions as a drive source for the piston drive mechanism 32. The gear 42 and the feed screw 44 convert the rotational motion of the motor 40 into a translational motion in the Z direction and transmit it to the carriage 46 . The carriage 46 is attached to the proximal end of the piston rod 30B and translates in the Z direction together with the piston body 30. As a result, when the motor 40 rotates, the piston body 30 moves in translation in the Z direction together with the carriage 46. In this way, the piston drive mechanism 32 can drive the piston body 30. Note that the configuration of the piston drive mechanism 32 is not limited to the above example, and any mechanism that moves the piston body 30 can be used.

[Configuration of valve mechanism 108]
Valve mechanism 108 manipulates the venting of gas within syringe 104 by changing the communication state of the interior space of sample dispensing assembly 100 . Valve mechanism 108 can be switched between a closed configuration that does not allow venting and an open configuration that allows venting. For example, valve mechanism 108 is a valve that can be electrically controlled and operated. Specifically, as shown in FIGS. 2 and 3, the valve mechanism 108 is configured as a solenoid valve, and includes a valve actuator 50 and a valve drive mechanism 70.

The valve actuator 50 switches the state of communication between the internal space of the sample discharge assembly 100 and the vent hole 20 by changing the arrangement of the valve mechanism 108 . Valve actuator 50 is attached to syringe 104. Specifically, as shown in FIGS. 2 and 3, the valve operating section 50 includes a first valve operating section 52 and a second valve operating section 54. The first valve operating section 52 and the second valve operating section 54 are arranged on the upper surface 12A of the manifold 12. The second valve actuator 54 is disposed on the upper surface 12A of the manifold 12 on the proximal side of the first valve actuator 52.

As shown in FIGS. 3 to 7, the first valve operating section 52 is provided with a valve body 60, a valve chamber 62, a valve seat 64, a fourth passage 66, and a fifth passage 68. The valve body 60 is provided to control the flow of fluid within the first valve operating section 52 . Valve body 60 is switchable between a closed configuration (FIG. 6) and an open configuration (FIG. 7). Specifically, the valve body 60 can be switched between a closed configuration and an open configuration by moving. The valve chamber 62 serves as an internal space of the first valve operating section 52 and accommodates the valve body 60 . The valve chamber 62 communicates with the gas chamber 22 and with the vent hole 20 open to the external space. The valve seat 64 contacts and supports the valve body 60 in the closed configuration, as shown in FIG.

As shown in FIGS. 5 and 6, the fourth passage 66 extends from the lower surface 52A of the first valve operating portion 52 to the valve chamber 62. The fourth passage 66 is formed to be continuous with the second passage 26 of the manifold 12 within the XY plane. That is, the second passage 26 and the fourth passage 66 together form a passage that communicates and connects the second chamber 22B of the manifold 12 and the valve chamber 62.

As shown in FIGS. 4 and 6, the fifth passage 68 has a shape extending from the lower surface 52A of the first valve operating portion 52 to the valve chamber 62 on the distal side (+Z) of the fourth passage 66. As shown in FIGS. 6 and 7, the valve seat 64 surrounds the outlet of the fifth passage 68 to the valve chamber 62. As shown in FIGS. The fifth passage 68 is formed to be continuous with the third passage 28 of the manifold 12 within the XY plane. That is, the third passage 28 and the fifth passage 68 together form a passage that communicates and connects the vent hole 20 of the manifold 12 and the valve chamber 62. However, as shown in FIG. 6, when the valve body 60 is in the closed configuration, the valve body 60 contacts the valve seat 64 and connects the valve chamber 62 and the fifth passage 68 (and thus the vent hole 20). cut off communication between As shown in FIG. 7, when the valve body 60 is in the open configuration, the valve body 60 is located away from the valve seat 64 and between the valve chamber 62 and the fifth passage 68 (and thus the vent hole 20). communication is allowed. That is, the valve body 60 blocks communication between the valve chamber 62 and the vent hole 20 in the closed configuration, and allows the valve chamber 62 and the vent hole 20 to communicate in the open configuration.

In this way, the valve actuator 50 has a closed configuration that blocks communication between the valve chamber 62 and the vent hole 20 and a closed configuration that allows communication between the valve chamber 62 and the vent hole 20. It has a valve body 60 that is movable. When the valve body 60 is in the closed configuration, an internal space is formed inside the sample ejection assembly 100 that communicates with the ejection port 102B, and this internal space is isolated from the external space except for a portion connected to the ejection port 102B. Closed. That is, when the valve body 60 is in the closed configuration, the discharge port 102B functions as the only outlet that communicates this internal space with the external space. This internal space is a space that includes the sample space 102A, the gas chamber 22, and the valve chamber 62, which communicate with each other. The piston 106 can change the volume of this internal space. Specifically, the piston 106 can change the volume of the gas chamber 22 in the internal space by sliding within the gas chamber 22. The valve actuator 50 can discharge the gas in the gas chamber 22 from the vent hole 20 by moving the valve body 60 from the closed configuration to the open configuration.

The valve drive mechanism 70 switches the open/closed state of the valve mechanism 108 by moving the valve body 60 within the valve chamber 62 between a closed configuration and an open configuration. Specifically, as shown in FIGS. 3, 6, and 7, the valve drive mechanism 70 includes a solenoid 72, a plunger 74 that moves under the action of the solenoid 72, and a coil spring 76 that biases the plunger 74. , has. Plunger 74 switches valve body 60 between a closed configuration and an open configuration as it moves under the action of solenoid 72 .

The solenoid 72 functions as a drive source for the valve drive mechanism 70. Specifically, as shown in FIGS. 6 and 7, the solenoid 72 includes a bobbin 80, a coil 82, and a coil cover 84. The bobbin 80 has a cylindrical shape with flanges at both ends, and functions as the base of the solenoid 72. Coil 82 is a metal wire wound around bobbin 80. The coil 82 generates a magnetic field therein when a current is applied thereto. The coil cover 84 is attached to the bobbin 80 so as to cover the coil 82 wound around the bobbin 80.

Plunger 74 is a cylindrical member made of a magnetic material such as iron or steel. When a current is applied to the coil 82 of the solenoid 72, a magnetic field is generated in the axial direction of the coil 82. When the solenoid 72 acts on the plunger 74, it applies a force to the plunger 74 in a direction away from the valve seat 64 (in the -Z direction here), as shown by the arrow in FIG. Plunger 74 has a reduced diameter section 86 at its distal end. The small diameter portion 86 is inserted into the through hole of the second valve operating portion 54 . The valve body 60 is attached to an operating end 88 that is the distal end of the small diameter portion 86 . Thereby, the plunger 74 can move the valve body 60 in the Z direction as it moves. That is, solenoid 72 applies a force to plunger 74 such that plunger 74 moves valve body 60 from a closed configuration to an open configuration. Thereby, when the solenoid 72 acts on the plunger 74, the plunger 74 moves the valve body 60 from the closed configuration to the open configuration. Note that the plunger 74 may be formed integrally with the valve body 60.

The coil spring 76 (an example of an “elastic member”) is housed in a housing hole formed in the bobbin 80 and abuts against the proximal end of the plunger 74 . The coil spring 76 biases the plunger 74 in the direction toward the valve seat 64 (here, +Z direction). Thereby, when no current is applied to the coil 82, the coil spring 76 can press the valve body 60 against the valve seat 64 via the plunger 74. That is, the coil spring 76 indirectly biases the valve body 60 in the direction from the open configuration to the closed configuration. On the other hand, when a current is applied to the coil 82 and the solenoid 72 is actuated, the plunger 74 moves in the proximal direction (-Z direction) against the elastic force of the coil spring 76. As a result, the valve body 60 separates from the valve seat 64, and the valve chamber 62 and the vent hole 20 communicate with each other.

Thus, when the solenoid 72 is not operating, the valve drive mechanism 70 positions the valve body 60 in the closed configuration by the biasing force of the coil spring 76. Thereby, the valve body 60 contacts the valve seat 64 and blocks communication between the valve chamber 62 and the vent hole 20. On the other hand, when the solenoid 72 is operating, the valve drive mechanism 70 moves the plunger 74 together with the valve body 60 by the magnetic force of the coil 82 . Thereby, the valve body 60 separates from the valve seat 64 and allows the valve chamber 62 and the vent hole 20 to communicate with each other. In this way, the valve drive mechanism 70 can switch the communication state between the valve chamber 62 and the vent hole 20 at desired timing. Thereby, the gas inside the sample discharge assembly 100 can be discharged from the vent hole 20 as needed.

As described above, the mode in which the valve body 60 switches from the open configuration to the closed configuration when the solenoid 72 is activated is preferable in terms of power consumption and the like because the solenoid 72 does not need to be activated during normal times when no venting is performed. However, the configuration of the valve drive mechanism 70 is not limited to the above example. For example, an aspect may be adopted in which the valve body 60 switches from the closed configuration to the open configuration when the solenoid 72 is activated.

<Configuration of positioning unit 200>
The positioning unit 200 positions and fixes the sample discharge assembly 100. Specifically, as shown in FIG. 1, the positioning unit 200 includes a pedestal 210 and a moving mechanism 220.

Pedestal 210 functions as a base for positioning unit 200. The moving mechanism 220 is attached to the base 210. Movement mechanism 220 moves sample ejection assembly 100 relative to pedestal 210 and fixes it in a predetermined spatial position and orientation. The moving mechanism 220 has a fixed portion 222 at one end thereof. The fixing part 222 holds the sample discharge assembly 100 and fixes the position and orientation of the sample discharge assembly 100 with respect to the moving mechanism 220.

The positioning unit 200 can fix the sample ejection assembly 100 at a predetermined position and orientation with respect to the target T. Thereby, the accuracy of the operation of the sample discharge assembly 100 injecting the sample S into the target T can be improved.

<Configuration of control unit 300>
Control unit 300 controls the operation of sample dispensing assembly 100. For example, control unit 300 electrically controls operation of at least one of piston 106 and valve mechanism 108. Specifically, as shown in FIGS. 1 and 8, the control unit 300 includes an operation section 310, a processing section 320, and a drive control section 330.

The operation unit 310 receives instructions for operating the sample discharge assembly 100 from a user. The operating section 310 may be provided integrally with the control device that constitutes the control unit 300, or a part or all of the operating section 310 may be provided separately from the control device. The operation unit 310 may be provided as a physical operation element such as a switch or a button on the operation panel of the control unit 300, or may be provided as a virtual operation element such as a button displayed on the display panel of the control unit 300. Alternatively, it may be in the form of a physical hand switch or foot switch that can be operated with the user's hands or feet. These may be used together to configure the operation section 310.

Specifically, the operating section 310 includes a piston operating section 312 and a valve operating section 314. In the example of FIG. 1, a piston operating section 312 and a valve operating section 314 are provided separately from the control device including the processing section 320 and the drive control section 330.

The piston operation unit 312 receives an operation instruction for operating the piston 106 from the user. For example, the piston operating unit 312 receives an operation from the user to move the piston 106 in the distal direction (+Z direction) or the proximal direction (−Z direction). For example, if the piston operating section 312 is a foot switch, the user can operate the piston 106 to move distally by stepping on the piston operating section 312 to compress the gas in the gas chamber 22. Can be done. Thereby, the user can discharge the sample S from the discharge tube 102 by increasing the internal pressure of the syringe 104 at a desired timing. The user can inject the sample S in the discharge tube 102 into the target T at a desired timing by operating the piston operating section 312 with the discharge tube 102 inserted into the target T.

The valve operation unit 314 receives an operation instruction for changing the open/closed state of the valve mechanism 108 from the user. For example, the valve operation unit 314 receives an operation instruction from the user to move the valve body 60 from the closed configuration to the open configuration. For example, when the valve operating section 314 is a foot switch, the user can operate the valve mechanism 108 by stepping on the valve operating section 314 to cause the valve chamber 62 and the vent hole 20 to communicate with each other. This allows the user to perform a venting operation on the gas chamber 22 at a desired timing.

The processing unit 320 performs predetermined arithmetic processing on the input information. For example, the processing section 320 executes a predetermined calculation process by executing a program recorded in the storage device of the control unit 300. Specifically, the processing unit 320 receives a user's operation instruction from the operation unit 310, and generates a control signal for the sample discharge assembly 100 based on the received operation instruction. Specifically, the processing unit 320 can generate a control signal for the piston drive mechanism 32 or the valve drive mechanism 70.

The drive control unit 330 controls driving of the components of the sample ejection assembly 100 based on the control signal generated by the processing unit 320. Specifically, the drive control section 330 includes a valve drive control section 332 and a piston drive control section 334.

The valve drive control unit 332 controls the drive of the piston drive mechanism 32 using a control signal generated based on the user's operation instruction received by the piston operation unit 312. Specifically, the valve drive control unit 332 drives the motor 40 according to the user's operation instructions.

The piston drive control unit 334 controls the drive of the valve drive mechanism 70 using a control signal generated based on the user's operation instruction received by the valve operation unit 314. Specifically, the piston drive control unit 334 drives the solenoid 72 according to the user's operation instructions.

The user can continuously inject the sample S into the target T by continuing to operate the piston operating section 312. Alternatively, upon receiving a user's operation, the piston operating unit 312 may control the piston drive mechanism 32 to continuously move the piston 106 until the next operation is received. Alternatively, the piston operation unit 312 may be configured to allow multiple types of operations, and may be set in advance to move the piston 106 continuously for a predetermined period of time when a predetermined operation is received.

When the piston 106 is continuously moving in this way, the user can drive the valve drive mechanism 70 to move the valve body 60 from the closed configuration to the open configuration by operating the valve operating section 314. I can do it. This allows the user to vent the gas in the gas chamber 22 at desired timing. Therefore, while the sample S continues to be injected into the target T from the discharge tube 102, the user can stop the injection of the sample S at a desired timing by operating the valve operating section 314.

The configuration of the operation unit 310, processing unit 320, drive control unit 330, etc. described above is based on the cooperation of the hardware configuration including the processor included in the control device, memory, storage, input/output interface, communication interface, bus interconnecting these, etc. It is a functional part that is realized by working.

According to the configuration of the sample discharging system 1 as described above, the internal pressure of the syringe 104 can be released by operating the valve at any timing, so the discharging operation of the sample S becomes easy. On the other hand, in a conventional sample discharge system that does not include a valve mechanism such as the sample discharge system 1, it is difficult to stop discharge from a discharge tube at an arbitrary timing. In particular, when the organ-forming region of the fetus is targeted T, the pressure within the body cavity of the fetus, which is composed of small compartments, is high. Therefore, in order to inject the sample S, it is necessary to increase the internal pressure of the syringe (that is, the discharge pressure of the discharge tube) until it exceeds the internal pressure of the target T. When the discharge pressure exceeds the internal pressure of the target T, the sample S begins to flow in, but once the flow begins, the compression of the air inside the syringe becomes uncontrollable. As a result, the discharge tube continues to inject the sample S until the internal pressure of the syringe is sufficiently reduced. As a result, there is a possibility that an excessive amount of the sample S will flow into the body cavity of the fetus beyond the target T (organ forming region). Such an overdose can be fatal to the fetus. As described above, when injecting the sample S into the target T having a high internal pressure, there is a particular need for a sample discharge system that can stop discharging the sample S at a desired timing. Furthermore, when cells are used as the sample S, the viscosity differs each time, so a sample discharging system that does not depend on the viscosity of the sample S and can be operated in a general manner is required.

In addition, in the conventional sample discharge system, a method was also used in which the discharge tube 102 was manually pulled out from the target T at the timing when the discharge of the desired amount of the sample S was completed. However, with this method, it was difficult to adjust the injection amount of sample S with sufficient accuracy. When the discharge tube 102 was pulled out from the target T, there was a possibility that the tip of the discharge tube 102 would damage the target T, or that air could accidentally enter the target T. In addition, the sample S that was not discharged often spilled out of the discharge tube 102 during withdrawal, and the sample S was often wasted.

In the sample discharge system 1 according to the present embodiment, by operating the valve mechanism 108, the internal space of the syringe 104 and the vent hole 20 can be brought into communication. Thereby, the internal pressure of the syringe 104, which has increased due to the syringe 104 discharging the sample S from the discharge tube 102, can be reduced to atmospheric pressure. As a result, it is possible to stop discharging the sample S at a desired timing. Thereby, the operability of the sample discharging device is improved, and the discharge amount of the sample S can be adjusted with high precision. The operation of quickly pulling out the discharge pipe 102 from the target T upon completion of discharge, as in the conventional art, is no longer necessary. Since the pulling operation can be performed carefully, the possibility of accidentally getting air into the fetus can be reduced. The possibility that the sample S remaining in the discharge tube 102 will spill from the discharge tube 102 can also be reduced.

After pulling out the discharge tube 102 halfway, it is also possible to puncture the target T again by changing the puncturing angle slightly. In this way, by performing multiple punctures while finely adjusting the insertion angle of the discharge tube 102, a wide range of transplantation to the target T becomes possible. If cells can be transplanted over a wide area to some extent even in a narrow space, it is advantageous in that the number of regenerated nephrons will increase. On the other hand, it is difficult at present to once pull out the entire discharge tube 102, refill it with cells, and puncture the target T again at the same angle or at a finely adjusted angle.

Further, by using the piston drive mechanism 32, the valve drive mechanism 70, and the positioning unit 200, various operations can be performed while the sample discharge assembly 100 is fixed to the target T. Therefore, the safety of the injection operation can be improved, especially for a vulnerable target T such as a fetus.

<Sample discharge method>
Next, with reference to FIG. 9, a sample discharging method using the sample discharging system 1 will be described.
FIG. 9 is a flowchart showing the flow of a sample discharging method using the sample discharging system 1 according to the embodiment.

First, in step S901, the user accommodates the sample S in the sample space 102A of the discharge tube 102. The sample S may be accommodated with the discharge tube 102 attached to the syringe 104, or the discharge tube 102 may be attached to the syringe 104 after the sample S is accommodated in the discharge tube 102. Note that by operating the valve mechanism 108 when the discharge tube 102 is attached to the syringe 104, the possibility that the sample S in the discharge tube 102 is pushed out from the discharge port 102B by the internal pressure of the syringe 104 when attached is reduced. Can be reduced.

Next, in step S902, the user inserts the discharge tube 102 containing the sample S into the target T with the discharge tube 102 attached to the syringe 104. Next, in step S903, when the user operates the piston operating section 312, the piston drive mechanism 32 causes the piston body 30 to slide in the distal direction (+Z direction) within the syringe body 10. By pushing the piston 106 inside the syringe 104 in this way, the internal pressure of the syringe 104 increases. At this time, the valve drive mechanism 70 is not operating, and the valve mechanism 108 blocks communication between the gas chamber 22 of the syringe 104 and the vent hole 20. When the internal pressure of the syringe 104 exceeds a certain value, the sample S in the discharge tube 102 is pushed out from the discharge port 102B. In this manner, in step S904, the user can inject a desired amount of the sample S into the target T from the discharge tube 102.

The user uses a marker attached to the discharge tube 102 to determine whether a desired amount of the sample S has been injected. When the injection of the desired amount of sample S is completed, in step S905, the valve drive mechanism 70 is activated by the user operating the valve operation unit 314. The valve drive mechanism 70 causes the gas chamber 22 of the syringe 104 and the vent hole 20 to communicate with each other by moving the valve body 60 from the closed configuration to the open configuration. As a result, a part of the gas compressed within the syringe 104 is discharged from the vent hole 20, and the valve chamber 62 and the external space communicate with each other, thereby reducing the internal pressure of the syringe 104 to a pressure equal to that of the external space. . As a result, injection of the sample S from the discharge tube 102 to the target T is stopped. Thereafter, in step S906, the user pulls out the discharge tube 102 from the target T. It is also possible to puncture the target T again with a slight change in the angle and perform additional injection into a region within the target T where cells have not been administered.

In this way, by operating the valve mechanism 108 to vent the gas in the syringe 104, the injection of the sample S can be stopped at a desired timing without stimulating the target T. This may improve the safety, efficiency, and implantation range of the sample injection operation.

<Method for manufacturing artificial tissue>
Next, a method for manufacturing an artificial tissue using the sample discharge system 1 will be explained. In the following, an example will be described in which the target T is a fetus of an organ-deficient animal, and the sample S is a progenitor cell of the organ.

In this embodiment, an artificial tissue is manufactured by transplanting progenitor cells (sample S) into the organ-forming region (target T) of a fetus of a mid-stage or later animal fetus with a developmental environment using a so-called fetal organ complementation method. . Specifically, first, the progenitor cells (sample S) housed in the discharge tube 102 are injected into the organ-forming region (target T) of the organ-deficient fetus by the above sample discharge method explained with reference to FIG. do. For example, the injection operation can be performed transuterinely. At this time, by using the above sample discharging method, a highly accurate and safe injection operation is possible. This causes the progenitor cells to be transplanted into the fetus within the mother's body. The fetus, into which organ progenitor cells have been transplanted, is then raised within the mother's body. This allows the injected cells to grow within the fetus' body. Regenerated artificial organs are obtained from the fetus after it is born from the mother's body, or from the inside of the fetus that has been raised within the mother's body for a certain period of time. In this way, artificial tissues can be manufactured from progenitor cells. Note that the target T is not limited to the organ-forming region within the fetus. For example, the target T may be an organ or tissue extracted from a fetus.

The manufactured artificial tissue can be used as an organ for transplantation in regenerative medicine. Alternatively, human organs grown in animals may be used for safety tests and efficacy demonstration tests in drug discovery. In safety testing of new drugs, there are limits to the extrapolation of animal experiments to humans, and species differences between humans and rodents have been reported in more than 400 renal tubular transporters involved in drug metabolism, especially in the kidney. There is. When an artificial kidney is manufactured using kidney progenitor cells as sample S, nephrotoxicity, which can only be verified with human kidneys, can be verified within the scope of animal experiments. Specifically, we will develop new preclinical models (safety testing) using humanized kidney mice, drug discovery for drug-induced kidney injury using humanized kidney mice, and methods for treating kidney failure that will replace dialysis. Development is expected.

<Modified example>
The internal flow path configuration of the sample discharge assembly 100 is not limited to the above example. The internal space of the sample discharge assembly 100 may be connected to the sample space 102A from which the sample S is discharged, and the vent hole 20 from which gas is discharged to the external space. The position of the vent hole 20 and the configuration of the passage connecting the gas chamber 22 and the vent hole 20 are not limited to the above example, and any arrangement can be used.

In the above example, the sample discharge assembly 100 has the discharge tube 102 having a tubular shape, but the configuration of the discharge section is not limited to this. For example, the discharge pipe 102 may have any shape such as a conical shape or a pyramid shape. The sample space 102A may have a curved shape, a bent shape, a branched shape, or the like. The discharge pipe 102 may have two or more discharge ports 102B. Sample delivery assembly 100 may have more than one delivery tube 102.

In the above example, the syringe 104 has a cylindrical shape, but the configuration of the gas storage section is not limited to this. The gas storage section may have any form as long as it has a gas flow path for pushing out the sample from the discharge section. For example, instead of the cylindrical syringe 104, a gas containing part having an arbitrary shape such as a box shape or a spherical shape can be used.

Although in the example above, the internal pressure of sample delivery assembly 100 is increased by inserting piston 106 into syringe 104, any pressure mechanism other than piston 106 can be used. For example, a movable wall provided inside the syringe 104 may be moved by any means such as manual operation by a user, a motor, pneumatic pressure, hydraulic pressure, etc., and the internal volume may be changed. The syringe 104 itself may be deformable to change its internal volume. The internal pressure of sample dispensing assembly 100 may be increased by means of supplying gas within sample dispensing assembly 100. In addition, the configuration of the pressurizing mechanism is not particularly limited.

In the above example, the closed configuration and the open configuration are switched by translating the valve body 60 using a solenoid valve, but the configuration of the valve mechanism 108 is not limited to this. For example, a drive mechanism such as a motor for moving the valve body 60 may be provided. An arbitrary type of operating section such as a switch or a handle may be provided so that the user can manually move the valve body 60. Furthermore, the form of the valve body 60 is not limited to the above example. For example, the valve body 60 may have a membrane shape that can be opened and closed. The valve body 60 may be a shutter mechanism provided at the entrance, exit, or intermediate point of the passage from the valve chamber 62 to the vent hole 20. The valve body 60 may have a deformable shape that closes the passage from the valve chamber 62 to the vent hole 20 in an expanded state and opens the passage in a contracted state. A valve body 60 having both of these configurations or a plurality of valve bodies 60 may be provided. In addition, the configuration of the valve is not particularly limited.

In the above example, the coil spring 76 indirectly biases the valve body 60 via the solenoid 72, but the configuration of the valve drive mechanism 70 is not limited to this. For example, the coil spring 76 may act directly on the valve actuator 50 to bias the valve body 60 in the direction from the open position to the closed position.

In the above example, the control unit 300 controls the operations of the piston drive mechanism 32 and the valve drive mechanism 70, but the objects to be controlled are not limited thereto. For example, the control unit 300 may change or adjust the arrangement of the sample discharge assembly 100, insert or withdraw the discharge tube 102 from the target T, etc. by controlling the movement mechanism 220 of the positioning unit 200. This may further automate the sample injection sequence.

The user's operations and judgments in the above description of the sample discharging method may be performed by the control device instead, using any sensor as appropriate.

Renal progenitor cells were transplanted into the kidney-developing region of a kidney-deficient fetus (mouse) on the 13.5th day of pregnancy using a sample discharge system (manufactured by Takasago Electric Industries, Ltd.) having the above configuration. FIG. 10 is a photograph of the nephrogenic region of a mouse into which renal progenitor cells were transplanted using the sample ejection system. In FIG. 10, the positions of transplanted renal progenitor cells are indicated by arrows. It was confirmed that this sample discharging system allows accurate adjustment of the amount of renal progenitor cells injected.

DESCRIPTION OF SYMBOLS 1... Sample discharge system, 100... Sample discharge assembly (sample discharge device), 102... Discharge pipe (discharge part), 102A... Sample space, 102B... Discharge port, 104... Syringe (gas storage part), 10... Syringe body, DESCRIPTION OF SYMBOLS 12... Manifold, 20... Vent hole (vent part), 22... Gas chamber, 106... Piston (pressurizing mechanism), 30... Piston body, 32... Piston drive mechanism, 108... Valve mechanism, 50... Valve actuation part, 60 ... Valve body, 62 ... Valve chamber, 64 ... Valve seat, 70 ... Valve drive mechanism, 72 ... Solenoid, 74 ... Plunger, 76 ... Coil spring (elastic member), 200 ... Positioning unit, 300 ... Control unit, 310 ... Operation section, 320...processing section, 330...drive control section, S...sample, T...target

Claims (15)

A sample dispensing system comprising a sample dispensing assembly dispensing a sample, the sample dispensing system comprising:
The sample ejection assembly includes:
a discharge section having a sample space and a discharge port for discharging the sample housed in the sample space to the outside;
a gas storage section having a gas chamber communicating with the sample space and capable of storing gas in the gas chamber;
a pressurizing mechanism that increases the pressure within the gas chamber;
a valve mechanism having a valve chamber communicating with the gas chamber and a valve body capable of switching between a closed configuration and an open configuration;
Equipped with
The valve chamber communicates with a vent portion open to external space,
The valve body blocks communication between the valve chamber and the vent section in the closed configuration, and allows communication between the valve chamber and the vent section in the open configuration.
Sample discharge system. The pressurizing mechanism compresses the gas in the gas chamber to push out the sample from the discharge port via the gas.
The sample discharge system according to claim 1. When the valve body is in the closed configuration, an interior space is formed within the sample discharge assembly that communicates with the discharge port, and the interior space is separated from the external space except for a portion that connects to the discharge port. is closed,
The sample discharge system according to claim 1 or 2. The pressurizing mechanism changes the volume of the internal space.
The sample discharge system according to claim 3. The valve mechanism includes a solenoid and a plunger that moves under the action of the solenoid,
The plunger switches the valve body between the closed configuration and the open configuration when moved by the action of the solenoid.
The sample discharge system according to claim 1 or 2. the valve body switches between the closed configuration and the open configuration by moving;
when the solenoid acts on the plunger, the plunger moves the valve body from the closed configuration to the open configuration;
The sample discharge system according to claim 5. the valve body switches between the closed configuration and the open configuration by moving;
The valve mechanism includes an elastic member that directly or indirectly biases the valve body in a direction from the open configuration to the closed configuration.
The sample discharge system according to claim 5. The pressurizing mechanism has a piston body that is at least partially inserted into the gas chamber, and a piston drive mechanism that moves the piston body within the gas chamber.
The sample discharge system according to claim 1 or 2. further comprising a control unit that electrically controls operation of at least one of the pressurizing mechanism and the valve mechanism;
The sample discharge system according to claim 1 or 2. A sample discharge device for discharging a sample from a discharge section having a sample space inside that can accommodate a sample,
a gas containing section coupled to the discharge section, the gas containing section having a gas chamber that communicates with the sample space when the gas containing section is coupled to the discharge section;
a pressurizing mechanism that increases the pressure within the gas chamber;
a valve mechanism having a valve chamber communicating with the gas chamber and a valve body capable of switching between a closed configuration and an open configuration;
Equipped with
The valve chamber communicates with a vent portion open to external space,
The valve body blocks communication between the valve chamber and the vent section in the closed configuration, and allows communication between the valve chamber and the vent section in the open configuration.
Sample discharge device. A method for discharging a sample from a discharge section having a sample space inside that can accommodate the sample, the method comprising:
accommodating a sample in the sample space;
pushing the sample out of the discharge part by increasing the internal pressure of the sample space;
switching a valve body capable of switching between a closed configuration and an open configuration from the closed configuration to an open configuration, the valve body blocking communication between the sample space and an external space in the closed configuration; communicating the sample space with an external space in an open configuration;
including,
Sample discharge method. further comprising the step of inserting the discharge part containing the sample into a target (excluding a human),
In the step of pushing out the sample from the discharge part, the sample is injected into the target.
The sample discharging method according to claim 11. the target is a fetus of a non-human animal;
The sample discharging method according to claim 12. The sample contains cells.
The sample discharging method according to any one of claims 11 to 13. A method for manufacturing an artificial tissue, the method comprising:
Inserting a discharge part containing cells in an internal sample space into a fetus of a non-human animal;
injecting the cells into the fetus by increasing the pressure of gas in the sample space to force the cells out of the outlet;
switching a valve body capable of switching between a closed configuration and an open configuration from the closed configuration to an open configuration, the valve body blocking communication between the sample space and an external space in the closed configuration; communicating the sample space with an external space in an open configuration;
pulling out the discharge part from the fetus;
growing the fetus injected with the cells in the mother's body;
including,
Method for manufacturing artificial tissue.

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