JP5053628B2 - PRESSURE DEVICE, PRESSURE METHOD THEREOF, PUMP DEVICE, AND CULTURE DEVICE - Google Patents

Description

The present invention relates to a structure of a pressurizing apparatus using a liquid, gel or the like as a pressure transmission medium, a pressurizing method thereof, a pump apparatus, and a culture apparatus. The pressurizing apparatus of the present invention is widely used in a pressure resistant durability apparatus used for pressure resistance measurement of materials and parts, a breeding experiment apparatus under a high pressure such as a deep sea organism, a culture apparatus for culturing living cells and living tissues, and the like.

  Biological cell culture has been performed in a culture dish under atmospheric pressure, but in recent years it has been proposed to cultivate in an environment that mimics the body, and articular cartilage and bone that are subject to pressure and load in the body. Attempts have been made to culture under pressure. In particular, in regenerative medicine, in order to be used for treatment, it is important to dispose of pressure chambers and fluid passage portions to prevent contamination of other people's genes and infection pathogens. The same can be said for the breeding of microorganisms and organisms, and the testing of materials that dislike contact with high-purity substances and other substances. For this purpose, it is required to simplify the structure as much as possible and reduce the cost.

  By the way, in order to create a high pressure, it is common to use a turbine pump, a gear pump, a plunger pump, or the like, and close a pressure-resistant container into which a fluid has been injected so that the inside of the container has a high pressure. Patent Documents 1 and 2 disclose pressure devices.

  In order to raise organisms under high pressure, seawater or fresh water is injected into the aquarium containing seawater or freshwater from the outside to increase the pressure in the aquarium. For example, this is the technique disclosed in Patent Document 3.

In the culture of living cells and tissues used for regenerative medicine, etc., culturing while applying pressure while imitating the body is good for maintaining phenotype, maximal expression, differentiation induction, proliferation, cell migration, mass transfer. It is said to bring about an effect, and a pressure culture apparatus (patent documents 4 and 5) has been proposed.
JP-A-7-327676 JP-A-9-151903 Japanese Patent Laying-Open No. 2005-6547 JP 2001-238663 A Special table 2004-512031

  By the way, in the pressure resistance / durability apparatus of Patent Document 1 or Patent Document 2 and the breeding experiment apparatus of Patent Document 3, the liquid around the object subjected to pressure is pressured by rotation of the turbine, reciprocation of the plunger, rotation of the gear motor, or the like. The pressure is increased. At this time, the lubricating oil for smoothly rotating the turbine, the reciprocating movement of the plunger, the rotation of the gear motor, and the like and the sealing material for preventing leakage may be dissolved or mixed in the liquid. The outflow and contamination of the sealing material will limit its application.

  In the culture of living cells and tissues disclosed in Patent Document 5, the cells are sealed in water in a pressurized chamber pressurized with a pump in order to avoid mixing other substances into the culture solution. There is a seal bag. Even when such a seal bag is used, there is a risk of invasion of germs or the like when exchanging the culture solution or transferring to another seal bag, and the work requires careful attention and is troublesome.

  In the culture apparatus disclosed in Patent Document 4, the pressurization mechanism and the culture chamber are isolated from each other, and the pressurization liquid and the culture solution are not in direct contact with each other, which is excellent in preventing contamination. However, when the pressurizing mechanism is complicated, the manufacturing cost becomes high. Moreover, in the liquid feeding apparatus which sends a culture solution, there exists a possibility of producing fine powder by abrasion of a seal | sticker part (O-ring).

  Regarding such problems, Patent Documents 1 to 5 do not disclose the problem, and do not disclose means for solving the problem.

  Accordingly, a first object of the present invention relates to a pressurizing apparatus that uses a liquid or the like as a pressure transmission medium, and separates the pressurizing side and the pressure receiving side to prevent contamination on the pressure receiving side.

  Specifically, the purpose is to realize a pressurization apparatus that can be applied to inexpensive pressurization culture or the like with a simple pressurization mechanism while preventing the entry of external germs and preventing the entry of impurities. .

  A second object of the present invention is to provide a pressurizing method for a pressurizing apparatus that prevents contamination on the pressure receiving side.

  A third object of the present invention is to realize a pump device that pumps a fluid using the pressurization device described above.

The fourth object of the present invention relates to a culture apparatus for culturing living cells and living tissues, and is to provide a culture apparatus using one or both of the aforementioned pressurization apparatus and pump apparatus.

Therefore, in order to achieve the first object, a first aspect of the present invention is a pressurizing device for transmitting pressure to a pressure receiving side by a pressure transmission medium, and a pressure side space in which the pressure transmission medium is accommodated. A pressure-receiving side space that receives pressure, a pressure-receiving membrane that is installed between the pressure-side space and the pressure-side space, and can be deformed according to the pressure transmission medium, and the pressure-side space is connected, and a cylinder portion internal volume that narrows the slave piston is inserted, is connected to the cylinder portion, and a pressure piston which slides move the follower piston in the cylinder portion, the pressurizing piston A piston mechanism having a stretchable membrane that seals between the pressure piston and the cylinder, and moves the driven piston by sliding the pressure piston. the movement of the piston Transmitted to the pressure receiving layer by the pressure transmitting medium is configured such that pressurizing the pressure receiving-side space side. With this configuration, the above object can be achieved.

  In order to achieve the first object, in the pressurizing apparatus, preferably, the pressure transmission medium may be a liquid or a gel.

In order to achieve the first object, in the pressurizing device, preferably, when a liquid is used for the pressure transmission medium, the pressurizing device includes a stopper that seals the pressurizing side space that contains the liquid , the movement of the slave piston, the slave piston may be configured you open plug against the plug.

In order to achieve the second object, a second aspect of the present invention is a pressurizing method of a pressurizing device that transmits pressure to a pressure receiving side by a pressure transmission medium, and includes a pressure side space portion including a pressure receiving film. The step of inserting the pressure transmission medium, the step of connecting the pressure side space portion in which the pressure transmission medium has been inserted, and the cylinder part in which the driven piston is inserted to reduce the internal volume , and the driven part in the cylinder part A piston mechanism portion is connected to the cylinder portion, and includes a pressure piston that moves the piston, and an elastic membrane that allows the pressure piston to slide and seals between the pressure piston and the cylinder portion. a step, sliding the pressure piston in the cylinder unit, the move the slave piston to transmit movement of the driven piston in the pressure-receiving layer by the pressure transmission medium, the pressure receiving side space pressurizable to A configuration including a that step. With this configuration, the above object can be achieved.

  In order to achieve the second object, in the pressurizing method of the pressurizing apparatus, preferably, the pressure transmission medium may be a liquid or a gel.

  In order to achieve the second object, in the pressurizing method of the pressurizing apparatus, preferably, when a liquid is used as the pressure transmission medium, a plug for sealing the pressurizing side space containing the liquid It is good also as a structure including the driven piston which can be moved by the said pressurizing piston in the said cylinder part, and opening the stopper piston against the said stopper.

In order to achieve the third object, a third aspect of the present invention is a pump device for transferring a fluid to be transferred according to an external pressure, a pressure side space portion in which a pressure transmission medium is accommodated, a pressure Pressure receiving side space for flowing the fluid to be transferred, a pressure receiving membrane installed between the pressure side space and the pressure receiving side space, and deformable according to the pressure transmission medium, and the pressure side is connected to the space portion, a cylinder portion internal volume that narrows the slave piston is inserted, coupled to said cylinder portion, and slid pressure piston for moving the follower piston in the cylinder unit, the A piston mechanism that includes a stretchable membrane that allows the pressure piston to slide and seals between the pressure piston and the cylinder, and moves the driven piston by sliding the pressure piston. , the driven piston The movement is transmitted to the pressure-receiving membrane by the pressure transmission medium, and the pressure-receiving side space is reduced or pressurized, and the fluid to be transferred is poured into the pressure-receiving space when the pressure is reduced. In the configuration, the fluid to be transported flows out from the pressure side space. With this configuration, the above object can be achieved.

In order to achieve the third object, in the pump device, preferably, the pressure receiving side space portion may include a valve that opens at the outlet portion when the pressure is applied, and a valve that opens at the inlet portion when the pressure is reduced. . Further, in the above pump device, when a liquid is used as the pressure transmission medium, the pump device includes a plug that seals the pressure side space that contains the liquid, and the driven piston is applied to the plug by the movement of the driven piston. It is good also as a structure which opens and opens.

In order to achieve the fourth object, a fourth aspect of the present invention is a culture apparatus including a pressurizing device that transmits pressure to a pressure receiving side by a pressure transmission medium, wherein the pressurizing device is configured to transmit the pressure. A pressure-receiving space that receives a medium; a pressure-receiving-side space that receives pressure; and a pressure-receiving film that is installed between the pressure-side and the pressure-receiving-side space and can be deformed according to the pressure transmission medium. If, coupled to said pressure side space, the cylinder portion internal volume that narrows the slave piston is inserted, is connected to the cylinder portion, and a sliding pressure for moving the driven piston in the cylinder portion A pressure piston, and a piston mechanism portion that includes a stretchable membrane that allows the pressure piston to slide and seals between the pressure piston and the cylinder portion , and the pressure piston slides to Move the driven piston , Transmits the movement of the driven piston in the pressure-receiving layer by the pressure transmitting medium is configured such that pressurizing the pressure receiving-side space side. With this configuration, the above object can be achieved.

In order to achieve the fourth object, a fifth aspect of the present invention is a culture device including a pump device that transfers a culture solution according to an external pressure, and the pump device contains a pressure transmission medium. A pressure-side space portion, a pressure-receiving side space portion that receives pressure and allows the culture medium to flow, and is disposed between the pressure-side space portion and the pressure-receiving side space portion, and is deformable according to the pressure transmission medium a pressure receiving membrane, is connected to the pressure side space, the cylinder portion internal volume that narrows the slave piston is inserted, is connected to the cylinder unit, moves the slave piston to slide within said cylinder portion comprising a pressure piston which, with a piston mechanism and a stretch film to seal between the permit sliding of pressurizing piston to and with the pressure piston the cylinder portion, the sliding of the pressure piston By said driven piston The moved, the vacuum or pressurized state the pressure receiving-side space side by transmitted to the pressure receiving layer by the pressure transmitting medium body movement of the driven piston, under decompression, pouring the culture liquid to the pressure receiving side space The culture solution is allowed to flow out from the pressure receiving side space during pressurization. With this configuration, the above object can be achieved.
In the culture apparatus, when a liquid is used as the pressure transmission medium, the culture apparatus includes a plug that seals the pressure side space that contains the liquid, and the driven piston is applied to the plug by the movement of the driven piston. It is good also as a structure which opens and opens.

  According to the present invention, the following effects can be obtained.

  (1) The pressure side and the pressure receiving side can be separated, and even if harmful substances and contaminants are generated on the pressure side, the pressure receiving side is not affected.

  (2) When a liquid is used for the pressure transmission medium, air can be prevented from being mixed in and pressure transmission can be improved.

  (3) Since residual air can be minimized, high pressure can be obtained even if the moving distance on the pressure mechanism side is reduced.

  (4) A liquid such as a culture solution on the pressure chamber side can be exchanged or replenished independently of the pressure side.

(5) The structure on the pressure mechanism side can be simplified, and the manufacturing cost can be reduced.

[First Embodiment]

  A first embodiment of the present invention will be described with reference to FIGS. FIG. 1 is a view showing a pressurizing device according to the first embodiment, FIG. 2 is an exploded perspective view showing a pressurizing device and a valve, and FIGS. 3 and 4 are a cylindrical portion and a plug of a pressure chamber portion. FIG. 5 is a view showing an example of a pressure sensor, FIG. 6 is a cross-sectional view showing a pressure piston portion, FIG. 7 is a view showing a diaphragm on the pressure piston side, and FIG. 8 is a pressure chamber portion. FIG. 9 is a view showing a pressurizing device in the initial state, FIG. 10 is a view showing release of sealing of the pressurizing side space of the pressure chamber, and FIG. The figure which shows the pressurization apparatus in a pressurization cancellation | release state, FIG. 12 is a figure which shows the pressurization apparatus in a pressurization state.

  As shown in FIGS. 1 and 2, the pressurizing device 2 is means for transmitting pressure applied from the pressurizing side to the pressure receiving side by a pressure transmission medium, and includes a pressure chamber section 4 and a pressurizing mechanism section 6. The pressure chamber unit 4 and the pressurizing mechanism unit 6 are detachable. The pressure chamber unit 4 is installed inside the incubator 8, and the pressurizing mechanism unit 6 is installed outside the incubator 8.

[Configuration of Pressure Chamber 4]

  The pressure chamber portion 4 includes a coupling portion 12 and a lid portion 14 with the main body portion 10 interposed therebetween. For example, a space 16 is formed in the main body 10 as a pressure-receiving side space that receives pressure from the coupling portion 12 side. In the space 16, for example, a culture solution 18 is stored as a fluid, and as a test piece, for example, A culture object 20 is accommodated. In this case, the culture target 20 is any one of a living cell, a living tissue, and a cell constituent in which cells are seeded on a culture substrate. The main body 10 is also provided with an inlet port 22 for allowing the culture medium 18 to flow into the space 16 and an outlet port 24 for allowing the culture medium 18 to flow out of the space 16. A first valve 28 is installed in the connected pipeline 26, and a second valve 32 is installed in the pipeline 30 connected to the outlet port 24. These valves 28 and 32 need to have a proof strength capable of withstanding the pressure applied to the pressure chamber section 4. The valves 28 and 32 are opened, the culture medium 18 is circulated and filled in the space 16, and the valves 28 and 32 are closed to confine the culture medium 18 in the space 16. In this state, the culture medium 18 and the culture object 20 Can be pressurized.

  A pressure receiving film 34 formed of a stretchable plastic film or other thin film is installed between the main body portion 10 and the coupling portion 12, and an O-ring portion 36 is integrally formed around the pressure receiving film 34. Is formed. The O-ring portion 36 seals between the main body portion 10 and the coupling portion 12. The main body 10 and the lid 14 are sealed by O-rings 42 and 44 inside and outside the standing wall 38 by engaging the standing wall 38 on the main body 10 side with the groove 40 on the lid 14 side.

  A space portion 46 is formed as a pressure side space portion on the coupling portion 12 side, and a cylindrical portion 48 is formed. The space portion 46 is a space extending between the coupling portion 12 and the pressure receiving film 34 and reaching the inside of the cylindrical portion 48, and is filled with a liquid, for example, water 50 as a pressure transmission medium. As shown in FIG. 3A, the cylindrical portion 48 is provided with a step 51 in the middle and a large-diameter portion 52 and a small-diameter portion 54. A cylindrical plug 56 (FIG. 1) is installed as a plug for opening and closing the small diameter portion 54 side of the cylindrical portion 48.

  The plug 56 is configured as shown in FIG. 3B. The diameter of the plug 56 is set smaller than the inner diameters of the small diameter portion 54 and the large diameter portion 52, and the plug 56 is attached to a groove portion 58 formed in the middle portion thereof. The outer diameter of the O-ring 60 is set larger than the inner diameter of the small-diameter portion 54. Further, an O-ring 66 is attached to the distal end portion of the cylindrical portion 48 in order to achieve close bonding with the pressurizing mechanism portion 6 side on the outside thereof, and a tapered surface 68 is formed on the inside thereof.

  When the plug 56 is fitted with the O-ring 60 to the small diameter portion 54 side of the cylindrical portion 48, the cylindrical portion 48 is closed by the plug 56 (FIG. 1), and the plug 56 is removed to the large diameter portion 52 side. Then, as shown in FIG. 4, a space 62 is formed between the outer surface portion of the plug 56 and the inner wall portion of the large diameter portion 52, and the water 50 flows. The space 46 is set to be larger than the maximum protrusion width of the pressure piston 64 on the pressure mechanism 6 side.

[Configuration of Pressure Mechanism 6]

  As shown in FIG. 1, the pressurizing mechanism unit 6 includes a cylinder unit 70, a connecting unit 72, and a piston mechanism unit 74. The connecting portion 72 is fixed to the cylinder portion 70 by a plurality of screws 76 as fixing means, for example, and is fixed to the wall portion 78 of the incubator 8 by, for example, screws 80 as fixing means. The connecting portion 72 is a means for connecting the pressure chamber portion 4 and the cylinder portion 70. In this embodiment, the connection part 72 penetrated by the through-hole 82 formed in the wall part 78 of the culture container 8 is inserted in the warehouse. A connecting cylinder part 86 having a connecting hole 84 communicating with the cylinder part 70 is formed in the connecting part 72, and the cylindrical part 48 on the pressure chamber part 4 side is inserted into the connecting cylinder part 86, and the pressure chamber part 4 are connected.

  A driven piston 88 that follows the pressurizing piston 64 is inserted into the cylinder portion 70, and the water 50 flows between the driven piston 88 and the inner wall portion 90 of the cylinder portion 70 and the driven piston 88 is allowed to move. A space 92 is formed. The driven piston 88 is a means for narrowing the volume of the inner wall portion 90 and is a lightweight rigid body, for example, a cylindrical body formed of synthetic resin. In this case, the diameter of the driven piston 88 is smaller than the inner diameter of the inner wall portion 90.

  The driven piston 88 is a volume narrowing member for filling the space formed between the plug 56 and the pressurizing piston 64 to eliminate the air in the space 92, and means for opening the plug 56. It is. The clearance formed between the driven piston 88 and the cylinder part 70 in the space 92 allows the driven piston 88 to move freely and is minimized within a range in which the pressure from the pressurizing piston 64 can be transmitted. In this case, since the clearance, that is, the space 92 is small, the remaining of unnecessary air can be suppressed, and the pressure in the cylinder portion 70 is increased to contribute to the increase in pressure.

  Further, a pressure sensor 94 for detecting the pressure applied to the inside of the inner wall portion 90 is attached to the cylinder portion 70, and the cylinder portion 70 is connected to the sensor portion 96 of the pressure sensor 94 through a detection hole 98 formed in the cylinder portion 70. Inside pressure is acting. The pressure sensor 94 converts the detected pressure into an electrical signal and outputs it. For example, the pressure sensor 94 includes a sensor unit 96 and a sensor circuit 97 as shown in FIG. Convert to This electrical signal V is output from the output terminal 99.

  A piston mechanism 74 is fixed to the back surface side of the cylinder part 70 by a plurality of screws 100 as fixing means. The piston mechanism part 74 includes a support cylinder part 102 and a pressurizing piston 64. A shaft support portion 104 is attached to the support cylinder portion 102, and the shaft portion 106 of the pressure piston 64 is slidably supported on the shaft support portion 104. A diaphragm retainer 109 as an operating protrusion is installed between the pressure piston 64 and the support cylinder portion 102 together with a diaphragm 108 which is a stretchable membrane. The diaphragm 108 seals the cylinder portion 70 and also pressurizes the pressure piston. 64 movements are allowed. A flange portion 110 is attached to the rear portion of the shaft portion 106, and a coil spring 112 is installed as an elastic member between the flange portion 110 and the shaft support portion 104, and a restoring force of the coil spring 112 is applied. Yes. A pressing point 113 is formed at the center of the flange portion 110, and a pressing force is applied to the pressing point 113 from an actuator 114 as a driving means. For example, a pressure is intermittently applied to the shaft portion 106 from the actuator 114 through the pressure point 113.

[Configuration of Pressurizing Piston 64]

  As shown in FIG. 6, the pressure piston 64 has a flange portion 116 formed on the shaft portion 106 side, and a stopper ring 118 is formed on the inner side of the support cylinder portion 102 with respect to the flange portion 116. The retraction position of the pressurizing piston 64 is determined by the flange portion 116 hitting the stopper ring 118.

  A diaphragm 108 is fixed to a front end portion of the pressurizing piston 64 by a fixing means such as an adhesive, and a cylindrical diaphragm retainer 109 is fixed with the diaphragm 108 interposed therebetween. An O-ring portion 120 is formed integrally with the peripheral portion of the diaphragm 108, and the O-ring portion 120 serves as a fixing means for the diaphragm 108 and also serves as an airtight holding means for the cylinder portion 70. Therefore, a concave portion 124 concentric with the through-hole 122 is formed in the front end surface portion of the support cylinder portion 102, and the O-ring portion 120 inserted and held in the recess 124 is formed between the support cylinder portion 102 and the cylinder portion 70. The O-ring part 120 is closely attached and sealed between the support cylinder part 102 and the cylinder part 70 by being interposed therebetween and fixed between the support cylinder part 102 and the cylinder part 70.

  Thus, the diaphragm 108 has its peripheral part fixed to the support cylinder part 102 side, its central part fixed to the pressure piston 64 side, and its intermediate part pressurized from the peripheral part of the diaphragm retainer 109. It is bent so as to cover the front end side of the piston 64, and is arranged so as to be movable between the periphery of the diaphragm retainer 109 and the pressure piston 64 and the support cylinder portion 102.

  As shown in FIG. 7, the diaphragm 108 is formed of a stretchable material and has a dome shape. The diaphragm 108 has a film portion 126, and a pressure piston 64 and a diaphragm retainer 109 are fixed to the center portion of the diaphragm 108. A boss portion 128 is formed, and an O-ring portion 120 is formed at an edge portion of the film portion 126 with a bending portion 130 interposed.

  Next, a pressurizing method of the pressurizing device 2 will be described. This pressurizing method includes an initial setting when the pressurizing apparatus 2 is assembled and a pressurizing operation during operation.

〔Initial setting〕

  After manufacturing each part of the pressurizing apparatus 2 shown in FIGS. 1 and 2, as shown in FIG. 8A, the pressure receiving film 34 is sandwiched between the main body part 10 and the coupling part 12, and the pressure receiving film 34. The O-ring part 36 in the periphery of the body is interposed between the main body part 10 and the coupling part 12 for sealing. In this state, the plug 56 is not attached. When steam sterilization (autoclave) at 100 [° C.] or higher is performed, steam sterilization may be performed here.

  As shown in FIG. 8B, the tubular portion 48 is sealed by filling the tubular portion 48 with water 50 and attaching a plug 56. FIG. 1 shows a state where the cylindrical portion 48 is filled with water 50 and sealed with a plug 56. When sterilization is performed by electron beam sterilization or gamma rays, sterilization may be performed here.

  The lid 14 is opened in the clean bench, the culture solution 18 and the culture object 20 are placed in the space 16 of the pressure chamber 4, so that no air remains in the space 16, and the main body 10 and the lid 14 are separated. The O-rings 42 and 44 are also sandwiched between them, the lid portion 14 is attached to the main body portion 10, and the main body portion 10 side of the pressure chamber portion 4 is sealed.

  As shown in FIG. 9, the pressure chamber portion 4 is attached to the connecting portion 72, and the cylinder portion 70 and the pressure chamber portion 4 are connected. Therefore, as shown in FIG. 10, when the flange portion 110 is moved in the compression direction of the coil spring 112 and the pressurizing piston 64 is moved to the cylinder portion 70 side, the pressurizing piston 64 hits the driven piston 88 and pressurizes. The driven piston 88 moves to the cylindrical portion 48 side together with the piston 64.

  The driven piston 88 moved to the cylindrical portion 48 side hits the plug 56, and as the driven piston 88 moves, the plug 56 moves to the large diameter portion 52 side of the cylindrical portion 48, and the sealing of the cylindrical portion 48 is released. The The water 50 held in the cylindrical part 48 by sealing the plug 56 flows from the cylindrical part 48 to the cylinder part 70 side through the connecting part 72 when the plug 56 is unsealed, and the cylinder part It moves into the space 92 between the inner wall 90 of the 70 and the driven piston 88. At this time, the pressure in the pressure chamber portion 4 is rising, but when the position of the pressurizing piston 64 is retracted, the pressure returns to the same pressure as the atmospheric pressure.

When the effective sectional area of the pressurizing piston 64 is S [m ² ] and the force applied to the pressurizing piston 64 is F (N, Newton), the pressure P is
P (Pa, Pascal) = F / S (1)
It becomes. This pressure is detected by a pressure sensor 94.

[Pressurization during operation]

  In the pressurizing device 2, when the pressurizing force from the actuator 114 is released and the pressurizing piston 64 is retracted by the restoring force of the coil spring 112, the state shown in FIG. 11 is obtained. FIG. 11 shows a state where the pressure piston 64 is pulled and the pressure drop point (0 [MPa]). In this case, if the valve 28 is a check valve and the valve 32 is switched to the closed state when the pressurizing piston 64 is retracted, the culture solution 18 is stored in the space 16 of the pressure chamber 4.

  From this state, as shown in FIG. 12, when the pressurizing piston 64 is moved to the cylinder part 70 side so as to be a set pressure, for example, 0.5 [MPa], the space 92 of the cylinder part 70 and the connecting part 72 is reduced. The water 50 moves to the pressure receiving membrane 34 side, and the pressure receiving membrane 34 expands. Due to the change in the pressure receiving film 34, the culture solution 18 in the space 16 in the pressure chamber 4 is pressurized, and at the same time, the culture object 20 is also pressurized.

  Periodic, intermittent, or continuous pressurization can be performed by alternately repeating such a pressurization state (FIG. 12) and its release state (FIG. 11).

  Then, if a necessary force is applied to the pressurizing piston 64 and moved in advance, it is fed back to the actuator 114 that applies the force to the pressurizing piston 64 using the output of the pressure sensor 94, and the force is controlled. The pressure of the chamber part 4 can be raised to an accurate pressure and maintained.

  It is also possible to create a repetitive pressure in which the change in pressure changes approximately in a sine curve by changing the increase and decrease in pressure two-dimensionally or trigonometrically and repeating this.

  When the culture time elapses, the pressure chamber unit 4 is removed from the pressurizing mechanism unit 6, moved to a clean bench, and the culture object 20 is taken out in the clean bench. The culture object 20 may be transplanted to a patient when used as regenerative medicine, and in other cases, it can be used for test studies such as culture state and tissue formation.

  As described above, examples of culturing living cells and tissues have been given, but other organisms may be used for the culture object 20. For example, if the culture solution 18 is seawater or fresh water and the culture object 20 is replaced with deep sea organisms, it can be applied to breeding deep sea organisms. Further, if the culture solution 18 is replaced with water or a liquid in a use environment and a part as a pressure test sample is used in place of the culture object 20, it can be applied to a pressure test of the part.

[Influence of residual air in cylinder part 70 and function of driven piston 88]

The influence of residual air in the pressurizing device 2 will be described using mathematical expressions. The compressibility of water during pressurization is 4.95 × 10 ⁻⁵ [cm ² / kg] to 4.35 when the temperature is in the range of 0 ° C. to 40 ° C. and the pressure is 100 atm or less. × 10 ⁻⁵ [cm ² / kg] For example, when 100 [cm ³ ] water is subjected to a pressure of 10 [kg / cm ² ], the amount of compression is about 0.045 [cm ³ ] (= about 45 μl), so it can be ignored. It is.

On the other hand, in the case of residual air,
P × V = P ₁ × V ₁ = constant (2)
It is. However, P is a pressure, V is a volume, P ₁ is an original pressure, and V ₁ is an original volume. For example, when the absolute pressure is 1 [kg / cm ² ], the air V having a volume of 100 [cm ³ ] is 10 [cm ³ ] and 1/10 when the absolute pressure is 10 [kg / cm ² ]. The volume is greatly reduced.

  When pressure is applied to the cylinder portion 70 in which water and air are mixed by the pressurizing piston 64, a desired pressure cannot be obtained unless the pressurizing piston 64 is moved by an amount corresponding to the compression of air. Therefore, it is desirable that the amount of air is as small as possible in the cylinder portion 70 to which pressure is applied.

  As a specific example, the calculation conditions are as follows.

The internal volume of the space 16 of the pressure chamber 4 is 100,000 [mm ³ ] (= 100 [cm ³ ])
Effective diameter of the pressure piston 64: 13.5 [mm]
Effective pressure receiving area (Ap) of the pressurizing piston 64 including the diaphragm 108: 143 [mm ² ]
Movable stroke (Lmax) of the pressure piston 64: 14 [mm]
Diameter of driven piston 88: 14.8 [mm]
Inner diameter of cylinder part 70 of pressurizing piston 64: 15 [mm]
Length of cylinder part 70: 70 [mm]
Diameter of the large diameter part 52 of the cylindrical part 48: 16.5 [mm]
Length of the large diameter portion 52 of the cylindrical portion 48: 16 [mm]
In this case, the length of the cylinder part 70 needs to be longer than the stroke of the pressurizing piston 64 so that the pressurizing piston 64 does not collide.
Pressurized pressure: 1.47 [MPa] = 15 [kg / cm ² ]
Atmospheric pressure: Po = 1 [kg / cm ² ].
It is assumed that the volume of residual air in the pressure chamber section 4 is 600 [mm ³ ]. (Assuming residual air equivalent to 20 mm in diameter and 2 mm in height)

  A) When the driven piston 88 is not installed in the cylinder portion 70 and the water 50 is not put into the space portion 46:

In this case, the volume Vp that the pressure piston 64 can compress is
Vp = Ap × Lmax (3)
Therefore, it becomes 2,002 [mm ³ ]. The air volume Vn of the space of the cylinder part 70 is 12,370 [mm ³ ], and the air volume Vs of the space part 46 is 3,421 [mm ³ ]. Assuming that the volume Vm of the remaining air in the pressure chamber section 4 is 600 [mm ³ ], the total volume Vo of air confined in the cylinder section 70 is
Vo = Vn + Vs + Vm (4)
Therefore, it is 16,391 [mm ³ ]. When the total volume Vo is compressed by the pressure piston 64, the compressed volume Vc is
Vc = Vo-Vp (5)
To 14,389 [mm ³ ]. (Liquid also contracts, but since it is very small and negligible compared to air, only the contraction of air is calculated.) The pressure Pc at this time is
Po × Vo = Pc × Vc (6)
Pc = Po × Vo / Vc (7)
From 1.14 [kg / cm ^2], since the pressure Pc is at an absolute pressure, when the relative pressure, the 0.14 [kg / cm ^2].

  From this, it becomes 0.014 [MPa] in an international unit, and when air is held in the pressurized portion, it means that the pressure cannot be increased.

  B) When the driven piston 88 is installed in the cylinder part 70 and the water 50 is put into the space part 46:

When the volume Vp that can be compressed by the pressure piston 64 is 2,002 [mm ³ ], the volume Vn of air existing in the space 92 of the driven piston 88 and the cylinder portion 70 is 327 [mm ³ ].

The air volume Vs of the space portion 46 is theoretically Vs = 0 when the water 50 is put into the space portion 46, but in reality, a slight amount of air is generated in the gap between the pressure receiving film 34 and the coupling portion 12. It is expected to remain. It is assumed that the volume Vs of this residual air is 600 [mm ³ ].

The volume Vm of the residual air in the pressure chamber section 4 is 600 [mm ³ ]. The total volume Vo of the air confined in the cylinder part 70 is 1,527 [mm ³ ] from the equation (4). In this case, the air volume Vo is smaller than the volume Vp that can be compressed by the pressurizing piston 64 and can be sufficiently pressurized or boosted.

C) When there is about 2,500 [mm ³ ] as residual air

In this case, 973 [mm ³ ] of air is added to the above-mentioned condition B), which means that the volume of water of 973 [mm ³ ] is reduced.

Assuming that the pressure is 15 [kg / cm ² ] (absolute pressure is 16 [kg / cm ² ]), the volume Vo of the air in the atmospheric pressure is 2,500 [mm ³ ]. Volume Vc is
Vc = Po × Vo / Pc (8)
To 156 [mm ³ ]. Shrinkage Vad is
Vad = Vo−Vc (9)
Therefore, it is 2,344 [mm ³ ]. This is because the compression of the pressurizing piston 64 is larger than the compressible volume of 2,002 [mm ³ ] even by air contraction alone, so pressurization of 1.5 [MPa] cannot be performed.

However, pressurization of about 3 [kg / cm ² ] (absolute pressure 4 [kg / cm ² ]) is possible. The volume Vo of the air in the atmospheric pressure is set to 2,500 [mm ³ ] as described above. The volume Vc after contraction is 625 [mm ³ ] from the equation (8), the contraction amount Vad is 1,875 [mm ³ ] from the equation (9), and the volume Vw of the water 50 in the pressure chamber section 4 is 101, 848 [mm ³ ], the contraction amount Vwd of the water 50 is 19 [mm ³ ], and the total contraction amount Vtd is 1,894 [mm ³ ]. From the compressible volume 2,002 [mm ³ ] of the pressure piston 64 Since it is small, pressurization of 3 [kg / cm ² ] is possible. Thus, even if it is impossible at high pressure, pressurization may be possible at low pressure. When the water 50 is pressurized with this pressure, the air in the pressure chamber section 4 gradually dissolves in the water 50 in the pressure chamber section 4 and the influence of the air is reduced. Air tends to dissolve in water at high pressure (Henry's law).

  When the pressure is increased from 15 minutes to 2 hours at this pressure and the air in the cylinder part 70 is dissolved in water and the volume is reduced, the water 50 or the culture solution 18 is supplied little by little, or if the pressure chamber part 4 is replaced. Since the air adhering to the inner wall and the culture object 20 does not exist as a gas, high pressure can be applied.

  In this way, rather than applying a high pressure, after pressurizing at a low pressure for a certain period of time and then applying a high pressure, even if air enters the water to be pressurized, Therefore, the influence of air can be avoided and a sufficiently strong pressure can be obtained.

  It is as follows if characteristic matters etc. are extracted and enumerated from the said embodiment. .

  (1) Since the space 16 on the pressure receiving side of the pressure chamber 4 is partitioned from the pressure side by the pressure receiving film 34, contamination on the pressure receiving side can be prevented. Even if harmful substances and contaminants are generated on the pressurizing mechanism 6 side, there is no effect on the pressure chamber 4 or the test piece.

  (2) Since the cylinder portion 70 and the piston mechanism portion 74 connected to the pressure chamber portion 4 are partitioned by the diaphragm 108, contamination of the internal space of the cylinder portion 70 can be prevented.

  (3) A liquid such as water 50 is sealed in the space that is the movable range of the pressurizing piston 64 so as to exclude gas that is harmful to the pressure rise, and the cylindrical portion 48 of the pressure chamber portion 4 is It is sealed.

  (4) The space volume on the pressure mechanism 6 side is set as small as possible. When a space is generated, liquid (solid) (driven piston 88) is placed in the space to eliminate air (gas) as much as possible.

  (5) After the pressure chamber unit 4 and the pressurizing mechanism unit 6 are coupled, the seal of the plug 56 is released, and the two spaces of the pressure chamber unit 4 and the pressurizing mechanism unit 6 are combined into one space. Is integrated.

  (6) The pressure is increased in two steps, such as by applying a low pressure to dissolve unnecessary air (gas) in the liquid and then increasing the pressure to the target pressure.

  (7) Since the pressurizing device 2 includes the inlet port 22 and the outlet port 24 in the pressure chamber section 4 and has a pump function, the liquid in the pressure chamber section 4 can be exchanged.

  (8) Since the operating space of the pressure piston 64 is filled with water 50 as a pressure transmission medium and excludes air, the pressure transmission performance is improved. Water that accumulates in the space in which the pressurizing piston 64 operates can be removed, and air that hinders pressurization can be eliminated. The water 46 is held in the space 46 by the plug 56 until the pressurizing mechanism 6 is connected. it can. Further, the water 50 can absorb air under high pressure, and as a result, the residual air can be minimized, so that the pressure piston 64 can obtain a high pressure increase with a small movement distance.

  (9) Since the two spaces partitioned by the plug 56 are integrated into one space after being connected to the pressurizing mechanism 6, pressure can be transmitted to the pressure chamber 4 by the operation of the pressurizing piston 64. The pressure sensor 94 can accurately detect the pressure.

  (10) Even if air is mixed in the pressurizing mechanism 6 or the pressure chamber 4, the pressurization value can be increased stepwise, and high pressure can be applied.

  (11) The liquid in the pressure chamber section 4 can be replaced or replenished automatically.

  (12) The complicated pressurizing mechanism 6 is unnecessary, and the cost of the pressurizing device 2 can be reduced.

[Second Embodiment]

  Next, a second embodiment of the present invention will be described with reference to FIG. FIG. 13 is a diagram illustrating a pressurizing apparatus according to the second embodiment. In FIG. 13, the same parts as those in FIG.

In the first embodiment, water 50 is used as an example of a liquid that is a pressure transmission medium. However, a gel may be used, and the second embodiment is a case where a gel is used as the pressure transmission medium. . A gel is a substance having a form intermediate between a liquid and a solid, and has a property having both flexibility and elasticity. Most of the gels are water-containing gels (hydrogels) containing a large amount of water and have a property of absorbing air . As described in the first embodiment, the phenomenon that the water 50 absorbs air under high pressure can occur in the gel as well. The gel maintains a certain shape, but its force is weak, it can easily break the molecular chain and can flow.

  Therefore, as shown in FIG. 13, instead of water 50 (FIG. 1), gelatin or the like may be gelled by a method such as injecting gelatin into the space 46 in a sol state and cooling. Thus, in the gel 150 injected into the space portion 46 on the coupling portion 12 side of the pressure chamber portion 4, the plug 56 necessary for the case of the water 50 (FIG. 1) is unnecessary. Since the gel 150 maintains a certain shape, it functions as the plug 56. Further, the holding power of the shape of the gel 150 is weak and can be easily broken by the pressurization by the sliding of the pressurizing piston 64 and flows. That is, the gel 150 functions as a plug 56 and water 50 (FIG. 1), and functions as a pressure transmission medium in the same manner as a liquid when the molecular chain of the gel 150 is broken.

  Further, since the mixed air dissolves in the gel 150 and functions to substantially eliminate the air in the space 46, the same pressure transmission function as that of the water 50 (FIG. 1) is achieved. Since other operations are the same as those in the first embodiment, the description thereof is omitted.

[Third Embodiment]

  Next, a third embodiment of the present invention will be described with reference to FIGS. FIG. 14 is a view showing a plunger pump according to a third embodiment, FIGS. 15 and 16 are views showing a pressure chamber portion, and FIG. 17 is a view showing a pressure side space portion of the pressure chamber portion and its closing. FIG. 18 is a diagram illustrating the release of the pressure side space portion of the pressure chamber portion, FIG. 19 is a diagram illustrating a pressure reduction state of the pressure side space portion of the pressure chamber portion, and FIG. 20 is a pressure side space of the pressure chamber portion. The figure which shows the pressurization state of a part, FIG. 21 is a figure which shows the modification by the side of a coupling | bond part.

  The plunger pump 200 is an example of a pump device, and includes a mechanism similar to the pressurizing device 2 of the first embodiment. The plunger pump 200 utilizes the piston mechanism and is transferred in accordance with the reciprocating motion of the piston mechanism. The fluid can be sent.

  When this plunger pump 200 is compared with the pressurizing device 2 of the first embodiment, the plunger pump 200 has a liquid 204 that is a fluid to be transferred from the inlet portion 202 in accordance with the rise and fall of the pressure in the pressure chamber portion 208. And the liquid 204 is discharged from the outlet 206. Since it is not necessary to accurately control the pressure increase / decrease, the pressure sensor 94 (FIG. 1) is unnecessary, and the driven piston 88 (FIG. 1) is also unnecessary.

  In the plunger pump 200, when the liquid 204 is drawn into the space where the pressure is high, it is necessary to overcome the high pressure and draw the amount equivalent to the moving volume of the pressure piston 256 more accurately. The residual air (gas) in the part must be reduced as much as possible. This is the same as the pressurizing device 2. Therefore, the plunger pump 200 includes a pressure chamber unit 208 and a pressurizing mechanism unit 210, which are detachable.

[Configuration of Pressure Chamber 208]

  The pressure chamber unit 208 includes a main body unit 212 and a coupling unit 214. A space portion 216 is formed as a pressure receiving side space portion in the main body portion 212, and the liquid 204 sucked from the inlet portion 202 is stored in the space portion 216, and the liquid 204 flows out from the outlet portion 206. Further, a check valve 218 as a valve is installed at the inlet 202 and a check valve 220 as a valve is installed at the outlet 206. The check valve 218 is provided with a ball 224 that opens and closes the passage 222, and a restoring force of a spring 226 is applied to the ball 224. Similarly, the check valve 220 is provided with a ball 230 for opening and closing the passage 228, and a restoring force of a spring 232 is applied to the ball 230. That is, the check valve 218 is opened by the suction force at the time of pressure reduction, and the check valve 220 is opened by the pressurizing force at the time of pressure increase.

  A pressure receiving film 234 formed of a stretchable plastic film or other thin film is installed between the main body part 212 and the coupling part 214, and an O-ring part 236 is integrally formed around the pressure receiving film 234. Is formed. The O-ring portion 236 provides a seal between the main body portion 212 and the coupling portion 214.

  As shown in FIGS. 14 and 15, a space portion 238 is formed as a pressure side space portion and a cylindrical portion 240 is formed on the coupling portion 214 side. The space portion 238 is a space extending between the coupling portion 214 and the pressure receiving film 234 and into the cylindrical portion 240, and is filled with a liquid such as water 258 as a pressure transmission medium. The tubular portion 240 is provided with a step 242 in the middle and a large-diameter portion 244 and a small-diameter portion 246. A cylindrical plug 248 is installed as a plug for opening and closing the cylindrical portion 240. The plug 248 is the same as that shown in FIG. The diameter of the plug 248 is set smaller than the inner diameters of the small diameter portion 246 and the large diameter portion 244, and the outer surface diameter of the O-ring 252 attached to the groove portion 250 formed in the middle portion of the plug 248 is the inner diameter of the small diameter portion 246. It is set larger. Therefore, when the plug 248 is fitted with the O-ring 252 on the small diameter portion 246 side, the tubular portion 240 is closed by the plug 248. When the plug 248 is detached, a space 254 is formed and the water 258 flows as shown in FIG. The space portion 238 is set to be larger than the maximum protrusion width of the pressure piston 256 on the pressure mechanism portion 210 side.

  In addition, an O-ring 260 is attached to the distal end portion of the tubular portion 240 in order to achieve tight bonding with the pressurizing mechanism portion 210 side on the outer side, and a tapered surface 262 is formed on the inner side.

[Configuration of Pressure Mechanism 210]

  As shown in FIG. 14, the pressurizing mechanism unit 210 includes a cylinder unit 264 and a piston mechanism unit 266. The cylinder portion 264 is attached by being fitted to the cylindrical portion 240, and the pressurizing piston 256 of the piston mechanism portion 266 is inserted therein. The aforementioned O-ring 260 is installed between the cylinder portion 264 and the cylindrical portion 240 and is sealed by this O-ring 260. The cylinder portion 264 is provided with an O-ring 268 and guide rings 270 and 272. The slidability and airtightness between the inner wall side of the cylinder portion 264 and the outer surface of the pressure piston 256 are provided. Is retained.

  A mechanism part cover 274 that surrounds the outside of the pressure piston 256 is attached to the cylinder part 264, and a shaft part 278 of the pressure piston 256 is attached to the guide cylinder 276 that protrudes to the rear side of the mechanism part cover 274. It is slidably supported. A flange portion 280 is attached to the shaft portion 278 of the pressure piston 256, and the flange portion 280 constitutes a stopper that prevents the pressure piston 256 from moving backward. A coil spring 282 is installed between the flange portion 280 and the outer wall portion of the cylinder portion 264, and a restoring force of the coil spring 282 is applied. Therefore, the pressure piston 256 is placed in a stopped state at a position where the flange portion 280 comes into contact with the mechanism portion cover 274 by the restoring force. In order to move the pressurizing piston 256 toward the pressure receiving membrane 234 side, a pressing force exceeding the restoring force of the coil spring 282 is required. Therefore, an actuator 284 that applies a force for moving the pressure piston 256 back and forth is installed behind the shaft portion 278 of the pressure piston 256.

  Next, a method for pressurizing the plunger pump 200 will be described.

  After assembling the components of the pressurizing mechanism 210, as shown in FIG. 17A, the pressure receiving film 234 is sandwiched between the main body 212 and the coupling portion 214, and the main body is held by the O-ring 236 of the pressure receiving film 234. The space between 212 and the coupling portion 214 is sealed. In the case of steam sterilization (autoclave) at 100 [° C.] or higher, only the pressure chamber 208 may be steam sterilized before plugging with the plug 248.

  As shown in FIG. 17B, after filling the space 238 of the cylindrical portion 240 of the coupling portion 214 with water 258, the cylindrical portion 240 is sealed with a plug 248. When sterilization is performed by electron beam sterilization or gamma rays, only the pressure chamber unit 208 is sterilized here.

  The pressure chamber part 208 is attached to the pressurizing mechanism part 210, and both are connected as shown in FIG.

  In this state, as shown in FIG. 18, the sealing of the tubular portion 240 is released by moving the pressure piston 256 toward the pressure receiving film 234. When the plug 248 is pushed by the pressure piston 256 and moves toward the large diameter portion 244, the seal of the tubular portion 240 is released, and the plug 248 moves into the space portion 238 of the tubular portion 240. At this time, a part of the water 258 in the tubular portion 240 reaches the O-ring 268 side that seals the pressure piston 256.

  When the force applied to the pressurizing piston 256 is released, the restoring force of the coil spring 282 acts between the cylinder portion 264 and the flange portion 280 as shown in FIG. Returns to the position where the flange portion 280 collides with the mechanism portion cover 274. In this state, the internal pressure of the pressure chamber portion 208 is reduced most, the space portion 216 of the pressure chamber portion 208 in the pressurized state is shifted to a reduced pressure state, and the check valve 218 is opened. At this time, the liquid 204 flows from the inlet portion 202 into the space portion 216 of the pressure chamber portion 208.

  Therefore, as shown in FIG. 20, when the pressurizing piston 256 is pushed in, the internal pressure of the pressure chamber portion 208 increases, the check valve 220 is opened, and the liquid 204 in the pressure chamber portion 208 flows out from the outlet portion 206. . Thus, the liquid 204 can be circulated by reciprocating the pressurizing piston 256.

  Even if a part of the sealing material such as the pressure piston 256 and the O-ring 268 or the oil applied to the sealing material is mixed into the water 258 by the sliding of the pressure piston 256, it is partitioned by the pressure receiving film 234. The liquid 204 such as a liquid is not affected at all, and the liquid 204 is not contaminated.

The space pressurized by the pressurizing piston 256 and the pressure chamber portion 208 are filled with a liquid and the contraction is negligibly small unless a gas such as air is contained. Therefore, the cross-sectional area of the pressurizing piston 256 is set to Sp and the stroke Lp. Then, the supply (circulation) liquid quantity Q is
Q = Sp × Lp (10)
The amount of supplied (circulated) liquid can be calculated from the operation stroke, and a flow sensor or the like is not necessary.

  In this embodiment, instead of forming the large diameter portion 244 of the cylindrical portion 240, as shown in FIGS. 21A, 21B, and 21C, a pair of groove portions 286 may be formed. Good. With such a configuration, the plug 248 is held in the cylindrical portion 240, and the water 258 in the space can be communicated with the pressure piston 256 side. 21A is a plan view showing the coupling portion 214, and FIG. 21B is a cross-sectional view taken along the line XXIB-XXIB in FIG. 21A, showing the plug position before release of sealing. FIG. 21C is a cross-sectional view taken along the line XXIC-XXIC of FIG. 21A and shows a state after release of sealing.

  Also in this embodiment, the water 258 as the pressure transmission medium may be composed of the gel 150.

[Fourth Embodiment]

  Next, a fourth embodiment of the present invention will be described with reference to FIG. FIG. 22 is a diagram illustrating a pressurizing device according to the fourth embodiment. In FIG. 22, the same parts as those in FIG. 1 or FIG.

  In the first embodiment, the pressurizing mechanism unit 6 includes a cylinder unit 70, a connecting unit 72, and a piston mechanism unit 74, and the pressurizing piston 64 sealed with the diaphragm 108 is used for the piston mechanism unit 74. However, the pressurizing mechanism 210 of the plunger pump 200 (FIG. 14) of the third embodiment is used for the pressurizing device 2.

  In such a configuration, the cylinder portion 264 is installed similarly to the plunger pump 200, and the space between the cylinder portion 264 and the pressurizing piston 256 is sealed using the O-ring 268. The sealing structure is simplified as compared with the pressure device 2 (FIG. 1) of the first embodiment, and the pressure resistance of the O-ring 268 is improved as compared with the case where the diaphragm 108 is used. . In addition, the driven piston 88 is omitted, and the plug 56 can be opened by the pressure piston 256. Since other operations are the same as those in the first embodiment, description thereof is omitted.

  In this embodiment as well, the water 50 as the pressure transmission medium may be composed of the gel 150.

[Fifth Embodiment]

  Next, a fifth embodiment of the present invention will be described with reference to FIGS. FIG. 23 is a diagram showing a culture device according to a fifth embodiment, FIG. 24 is a perspective view showing a pressurization device, a plunger pump, and a closing valve of the culture device, and FIG. 25 is a culture completion after the culture unit is assembled. FIG. 26 and FIG. 27 are flowcharts showing the culture process. 21 and 22, the same reference numerals are given to the same portions as those in FIGS.

  As shown in FIGS. 23 and 24, the culture apparatus 300 combines the pressurizing apparatus 2 (FIG. 1) and the plunger pump 200 (FIG. 14) to supply and pressurize the culture medium that enables pressurization and liquid feeding. Configure the device.

  The culture apparatus 300 includes a culture chamber 302, a mechanism unit 304, and a control device 306. The culture chamber 302 is provided with the pressure chamber section 4 of the pressurizing device 2, the pressure chamber section 208 of the plunger pump 200, the culture medium tank 308, the closing valve 310 and the waste liquid tank 312. The culture medium tank 308, the plunger pump 200, The pressure device 2 and the closing valve 310 are connected in series by a tube 314, and the culture solution 316 that has flowed from the culture solution tank 308 to the plunger pump 200, the pressurization device 2 and the closing valve 310 flows to the waste solution tank 312.

  In this case, as shown in FIG. 24, the outlet port 206 of the plunger pump 200 is connected to the inlet side port 22 of the pressure chamber portion 4 of the pressurizing device 2, and the closing valve is connected to the outlet side port 24 of the pressure chamber portion 4. The inlet part 318 of 310 is connected. These constitute the culture unit unit 320. Since the pressure chamber section 4 is sealed with the pressure receiving film 34, it is separated from the pressurizing mechanism section 6, and the sealing performance of the space section 16 is not deteriorated, but the pressurizing mechanism section 6 is connected to the culture unit 320. They may be moved together.

  As a modification of this embodiment, the culture solution 316 that has passed through the closing valve 310 may be returned to the culture solution tank 308 to circulate the culture solution 316.

  The mechanism unit 304 is provided with a pressurizing mechanism unit 6 of the pressurizing device 2, a pressure sensor 94, an actuator 114 thereof, a pressurizing mechanism unit 210 of the plunger pump 200, an actuator 284 thereof, and a driving unit 322 of the closing valve 310. .

  The control device 306 is configured by a computer, and controls a control unit 324 that controls operation according to an input, an operation switch 326, pressure intensity, pressurization cycle, pressurization time, liquid supply amount, supply cycle, and the like. Are input, and a display device 330 that displays the input, actual pressure value, and the like.

[Procedure for supplying culture medium, pressurizing, etc.]

  As shown in FIG. 25, the culture unit 320 is assembled (step S1). That is, the plunger pump 200 and the closing valve 310 are connected to the pressure chamber unit 4 to assemble the culture unit 320. Since it is a portion where pressure is applied, it is made of a material having sufficient pressure resistance and sealed. A culture medium tank 308 is connected to the plunger pump 200 through a tube 314. A tube 314 is also connected to the outlet side of the closing valve 310 so that the drained liquid is guided to the waste liquid tank 312. The connected culture unit 320 performs sterilization (step S2).

  The culture solution 316 is put in the culture solution tank 308 (step S3), and the culture object 20 is installed in the space 16 of the pressure chamber 4 (step S4). The culture target 20 is, for example, a living cell or a living tissue. The lid part 14 is attached (step S5), the space part 46 of the pressure chamber part 4 is sealed with the plug 56, and the outside air is shut off (step S6). Steps S3 to S6 are performed in a clean bench.

  The culture unit 320 is connected to the mechanism unit 304, that is, the pressure chamber unit 4 is attached to the pressurizing mechanism unit 6, and the pressure chamber unit 208 of the plunger pump 200 is connected and fixed to the pressurizing mechanism unit 210. Is connected to the drive unit 322 (step S7). The plug 56 is unsealed (step S8), and the pressure is increased or decreased by the pressurizing piston 64, the closing valve 310 is opened, the actuator 284 is operated, and the culture solution 316 is fed into the pressure chamber section 4 (step S9). The culture solution 316 may be filled when the culture object 20 is added. When the medium 316 is filled with the culture solution 316 from the plunger pump 200 to the closing valve 310, and the air remains almost completely, the closing valve 310 is closed. The actuator 114 of the pressurizing mechanism 6 is operated to pressurize to the set pressure. When pressure is periodically applied, the pressurizing piston 64 is reciprocated by the actuator 114 and is repeatedly changed at a set cycle between the set pressures. In addition, the culture solution 316 is supplied into the space 16 of the pressure chamber 4 at a set cycle, and is circulated or exchanged. When pressurization for a set period for culturing is completed (step S10), the culturing unit 320 is removed from the mechanism unit 304 (step S11). Steps S7 to S11 are performed in the culture apparatus 300.

  The lid 14 of the pressure chamber 4 is opened in the clean bench, and the culture object 20 is taken out (step S12). The culture state is inspected (step S13), and if the object to be cultured 20 satisfies the conditions for transplantation or the like, the test is accepted (step S14). It is stored in a container so that it does not exist, or it is returned again to the pressure chamber part 4 and the lid part 14 is closed, sent to the operating room, and used for processing such as transplantation (step S15).

  Next, the pressurizing device 2, the plunger pump 200, and the closing valve 310 for the culture process of the culture device 300 will be described with reference to FIGS. In FIGS. 26 and 27, reference signs A, B, and C indicate connecting portions between the flowcharts.

  The processing of the culture apparatus 300 is as follows. As shown in FIG. 26, the power is turned on (step S21), the setting display (step S22), the culture unit 320 is installed in the mechanism unit 304 (step S23), and the culture conditions are input (pressure pattern pressure value, pressure cycle, liquid feeding). Volume, incubation time) (step S24), setting value display (step S25), operation switch 326 is turned on (step S26), closing valve 310 is opened (step S27), pressure chamber section 4 and actuator 114 of plunger pump 200 284 operates in the pressurizing direction (step S28), sealing of the pressure chamber 4 and the plugs 56 and 248 of the plunger pump 200 is released (step S29), and the actuators 114 and 284 operate in the depressurizing direction (step S30). Stop the actuators 114 and 284 (step S31) and close the closing valve 310. Step S32) via, it is determined set pressure pattern (step S33). In continuous pressurization, the actuator 114 stops at the pressurization position (step S34), in periodic pressurization, the actuator 114 reciprocates (step S35), and in intermittent pressurization, the actuator 114 reciprocates intermittently (step S35). S36) is executed.

  As shown in FIG. 27, it is determined whether or not the pressure value is normal (step S37). When a certain time has passed (step S38), the pressurization operation is stopped (step S39), and the closing valve 310 is opened (step S40). The actuator 284 operates in accordance with the liquid supply amount (step S41), liquid supply (step S42), operation status display (pressure value, liquid supply amount, elapsed time, etc.) (step S43), pressure value, motor, etc. It is determined whether or not the operation state of the load on the drive unit 322, the control device 306, etc. is normal (step S44). When the culture operation time has elapsed (step S45), the actuator 114 is set to the pressure = 0 position (step S46). The origin position (step S47), the closing valve 310 is closed (step S48), the culture end display (step S49), the power is turned off (step S48). -Up S50) in the process is completed.

  By the way, when the pressure value is not normal in the determination of step S37, for example, it is determined whether or not it is within the third time (step S51). If it is within the third time, the pressure correction by the actuator 114 is performed. Is performed (step S52), and then the process returns to step S37.

  In the determination in step S51, if the predetermined number of times is not within the third time, for example, the pressurization operation is stopped, the actuator 114 is stopped at the pressure: 0 position, the closing valve 310 is opened (step S53), an alarm (step S54), and a warning Sound generation is performed (step S55).

  If it is determined in step S44 that there is an abnormality, the operation is stopped (step S56), an alarm (step S57), and a warning sound is issued (step S58).

  The features of the culture apparatus 300 and the culture method using the culture apparatus 300 are listed as follows.

  (1) Contamination of the culture object 20 can be prevented, and the pressure chambers 4 and 208 can be easily pressurized. Cost can be reduced with a simple configuration. Increased certainty of air exclusion.

  (2) If the gel 150 is used instead of the waters 50 and 258, it functions as a stopper while maintaining the same function as the liquid. Moreover, if it is a gel, a seal | sticker can be cancelled | released easily.

  (3) The force of the pressurizing piston 64 can be transmitted to the pressure chamber section 4 as a pressure, and high-pressure pressurization is easy.

(Other embodiments)

  In the above embodiment, water 50, 258 or gel 150 is used as the liquid as the pressure transmission medium, but oil may be used.

  In the fifth embodiment, when a gel is used as the pressure transmission medium, it is not necessary to seal the plug of the pressurizing device 2 or the plunger pump 200. Therefore, in the flowchart (FIG. 25), the plug is sealed. In the release (step S8) and the flowchart (FIG. 26), the plug seal release (step S29) becomes unnecessary.

As described above, the most preferable embodiment and the like of the present invention have been described. However, the present invention is not limited to the above description, and is described in the claims or disclosed in the specification. It goes without saying that various modifications and changes can be made by those skilled in the art based on the above gist, and such modifications and changes are included in the scope of the present invention.

The present invention relates to a pressurizing device that uses a liquid as a pressure transmission medium, and separates the pressurizing side and the pressure-receiving side to prevent contamination on the pressure-receiving side, preventing invasion of external germs and preventing contamination of impurities. However, a simple pressurization mechanism can be applied to inexpensive pressurization culture and the like, a pressurization method of a pressurization device that prevents contamination on the pressure receiving side, a pump device that pumps fluid using the pressurization device, a living cell, Regarding a culture apparatus for culturing a living tissue, a culture apparatus using one or both of the pressurization apparatus and the pump apparatus described above can be provided and useful.

It is a figure which shows the pressurization apparatus which concerns on 1st Embodiment. It is a disassembled perspective view which shows a pressurization apparatus and a valve | bulb. It is a figure which shows the cylindrical part and plug of a pressure chamber part. It is a figure which shows the cylindrical part and plug of a pressure chamber part. It is a figure which shows an example of a pressure sensor. It is sectional drawing which shows a pressurization piston part. It is a figure which shows the diaphragm by the side of a pressurization piston. It is a figure which shows a pressure chamber part and a plug. It is a figure which shows the connection of a cylinder part and a pressure chamber part. It is a figure which shows the movement to the cylinder part of a pressurization piston. It is a figure which shows the state which retracted the pressurization piston. It is a figure which shows a pressurization state. It is a figure which shows the pressurization apparatus which concerns on 2nd Embodiment. It is a figure which shows the plunger pump which concerns on 3rd Embodiment. It is a figure which shows a pressure chamber part. It is a figure which shows a pressure chamber part. It is a figure which shows the pressurization side space part of a pressure chamber part, and its closure. It is a figure which shows the closure cancellation | release of the pressurization side space part of a pressure chamber part. It is a figure which shows the pressure_reduction | reduced_pressure state of the pressure receiving side space part of a pressure chamber part. It is a figure which shows the pressurization state of the receiving pressure side space part of a pressure chamber part. It is a figure which shows the modification by the side of the coupling | bond part of a pressure chamber part. It is a figure which shows the pressurization apparatus which concerns on 4th Embodiment. It is a figure which shows the culture apparatus which concerns on 5th Embodiment. It is a disassembled perspective view which shows the pressurization apparatus, plunger pump, and closing valve of a culture apparatus. It is a flowchart which shows the process from the assembly of a culture apparatus to completion | finish of culture | cultivation. It is a flowchart which shows a culture | cultivation process. It is a flowchart which shows a culture | cultivation process.

Explanation of symbols

2 Pressurizing device 4 Pressure chamber 6 Pressure mechanism 16 Space (pressure receiving space)
46 Space (Pressure side space)
34, 234 Pressure receiving membrane 50, 258 Water 56, 248 Plug 64, 256 Pressure piston 70, 264 Cylinder portion 88 Driven piston 200 Plunger pump 300 Culture device 18, 316 Culture solution 320 Culture unit

Claims (12)

A pressurizing device that transmits pressure to a pressure receiving side by a pressure transmission medium,
A pressure-side space in which the pressure transmission medium is accommodated;
A pressure-receiving side space that receives pressure;
A pressure receiving membrane that is installed between the pressure side space and the pressure receiving side space and is deformable according to the pressure transmission medium;
Is connected to the pressure side space, and a cylinder portion that internal volume is narrowed slave piston is inserted,
A pressure piston connected to the cylinder portion and sliding to move the driven piston within the cylinder portion ; allowing the pressure piston to slide; and sealing between the pressure piston and the cylinder portion A piston mechanism comprising a stretchable membrane ,
The driven piston is moved by sliding of the pressure piston, the movement of the driven piston is transmitted to the pressure receiving film by the pressure transmission medium, and the pressure receiving side space portion side is pressurized. Pressurizing device. The pressurizing device according to claim 1.
The pressurizing device, wherein the pressure transmission medium is a liquid or a gel. The pressurizing device according to claim 1 or 2,
When using a liquid to the pressure transmission medium, comprising a plug which seals the pressure side space portion containing the liquid, by the movement of the slave piston, the opening plug to isosamples against the slave piston to said stopper A pressurizing device characterized. A pressurizing method of a pressurizing device for transmitting pressure to a pressure receiving side by a pressure transmission medium,
Placing the pressure transmission medium in a pressure side space provided with a pressure receiving membrane;
Connecting the pressurizing side space containing the pressure transmission medium and a cylinder part into which a driven piston is inserted to reduce the internal volume ;
A piston mechanism comprising: a pressure piston that moves the driven piston in the cylinder portion; and a stretchable membrane that allows the pressure piston to slide and seals between the pressure piston and the cylinder portion. Connecting to the cylinder part;
A step of sliding the pressure piston into the cylinder portion, moving the driven piston , transmitting the movement of the driven piston to the pressure receiving film by the pressure transmission medium, and enabling the pressure receiving side space portion to be pressurized;
The pressurizing method of the pressurizing apparatus characterized by including. In the pressurization method of the pressurization apparatus of Claim 4 ,
The pressurizing method of the pressurizing apparatus, wherein the pressure transmission medium is a liquid or a gel. In the pressurization method of the pressurization apparatus of Claim 4 ,
When a liquid is used as the pressure transmission medium, the pressure transmission medium includes a stopper that seals the pressurizing side space, and the cylinder includes a driven piston that can be moved by the pressurizing piston. A pressurizing method for a pressurizing device, comprising the step of opening the cap against the stopper. A pump device that transfers a fluid to be transferred according to an external pressure,
A pressure side space in which the pressure transmission medium is accommodated;
Pressure receiving side space that receives pressure and flows the fluid to be transferred; and
A pressure receiving membrane that is installed between the pressure side space and the pressure receiving side space and is deformable according to the pressure transmission medium;
Is connected to the pressure side space, and a cylinder portion that internal volume is narrowed slave piston is inserted,
A pressure piston connected to the cylinder portion and sliding to move the driven piston within the cylinder portion ; allowing the pressure piston to slide; and sealing between the pressure piston and the cylinder portion A piston mechanism comprising a stretchable membrane ,
The driven piston is moved by sliding of the pressure piston, the movement of the driven piston is transmitted to the pressure receiving film by the pressure transmission medium, and the pressure receiving side space portion side is depressurized or pressurized, The pump device according to claim 1, wherein the fluid to be transferred is poured into the pressure-receiving side space when the pressure is reduced, and the fluid is caused to flow out from the pressure-receiving space when the pressure is applied. The pump device according to claim 7 ,
The pressure receiving side space includes a valve that opens at the outlet when the pressure is applied, and a valve that opens at the inlet when the pressure is reduced. The pump device according to claim 7 or 8,
In the case of using a liquid as the pressure transmission medium, a plug is provided for sealing the pressurizing side space containing the liquid, and the driven piston is opened against the plug by the movement of the driven piston. The pump device. A culture device comprising a pressurizing device that transmits pressure to the pressure receiving side by a pressure transmission medium, wherein the pressurizing device is
A pressure-side space in which the pressure transmission medium is accommodated;
A pressure-receiving side space that receives pressure;
A pressure receiving membrane that is installed between the pressure side space and the pressure receiving side space and is deformable according to the pressure transmission medium;
Is connected to the pressure side space, and a cylinder portion that internal volume is narrowed slave piston is inserted,
A pressure piston connected to the cylinder portion and sliding to move the driven piston within the cylinder portion ; allowing the pressure piston to slide; and sealing between the pressure piston and the cylinder portion A piston mechanism comprising a stretchable membrane ,
The driven piston is moved by sliding of the pressure piston, the movement of the driven piston is transmitted to the pressure receiving film by the pressure transmission medium, and the pressure receiving side space portion side is pressurized. Characteristic culture device. A culture device comprising a pump device for transferring a culture solution according to an external pressure, wherein the pump device is
A pressure side space in which the pressure transmission medium is accommodated;
Pressure-receiving side space that receives pressure and flows the culture solution;
A pressure receiving membrane that is installed between the pressure side space and the pressure receiving side space and is deformable according to the pressure transmission medium;
Is connected to the pressure side space, and a cylinder portion that internal volume is narrowed slave piston is inserted,
A pressure piston connected to the cylinder portion and sliding to move the driven piston within the cylinder portion ; allowing the pressure piston to slide; and sealing between the pressure piston and the cylinder portion A piston mechanism comprising a stretchable membrane ,
Wherein the moving the slave piston by sliding the pressurizing piston, the reduced pressure or pressurized state the pressure receiving-side space side movement of the driven piston is transmitted to the pressure receiving layer by the pressure transmission medium body The culture apparatus is configured to flow the culture solution into the pressure receiving side space during decompression and to flow out the culture solution from the pressure receiving side space during pressurization. The culture apparatus according to claim 10 or 11,
In the case of using a liquid as the pressure transmission medium, a plug is provided for sealing the pressurizing side space containing the liquid, and the driven piston is opened against the plug by the movement of the driven piston. Culture apparatus.

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