CN215288827U

CN215288827U - Perfusion culture system

Info

Publication number: CN215288827U
Application number: CN202120642481.1U
Authority: CN
Inventors: 王玄
Original assignee: Shanghai Ruiyu Biotech Co Ltd
Current assignee: Shanghai Ruiyu Biotech Co Ltd
Priority date: 2021-03-30
Filing date: 2021-03-30
Publication date: 2021-12-24
Anticipated expiration: 2031-03-30

Abstract

The utility model discloses a perfusion culture system, including cultivateing the board, be provided with a plurality of cultivation holes on the cultivation board, the below of cultivateing the board is provided with the flexible tube, cultivates the hole and passes through first intercommunicating pore and flexible tube intercommunication, and the below of flexible tube is provided with a plurality of shutoff pieces, and shutoff piece selectivity is close to and supports tight flexible tube, and the shutoff piece is adjacent to first intercommunicating pore. The blocking piece is arranged below the flexible pipe and located outside the flexible pipe, so that a pipeline device and a control device of the perfusion culture system can be separated, the flexible pipe and the blocking piece respectively belong to the pipeline device and the control device, the structure of the perfusion culture system is simplified, and the operation difficulty of high-flux perfusion culture is reduced; through setting up the shutoff piece and being close to first through-hole, when the shutoff piece is close to and supports tight flexible pipe, under the extrusion of the lower surface of culture plate and shutoff piece terminal surface, the flexible pipe is compressed tightly, and the shutoff effect has been realized to the shutoff piece.

Description

Perfusion culture system

Technical Field

The utility model relates to a biomedical field especially relates to a perfusion culture system.

Background

Organoids are three-dimensional tissues, and conventional static culture causes many problems such as frequent contamination by liquid change, inability to simulate in vivo circulation environment, and the like. Perfusion culture is a relatively ideal three-dimensional culture mode in which a culture solution can flow in a culture chamber under the driving action of an external force, nutrients can be continuously updated, fluid shear force is provided, and introduction of exogenous substances (such as medicines) is facilitated. Based on the excellent functions of organoids, a large number of biotechnology service companies start to cooperate with hospitals and doctors to perform organoid culture and drug analysis, the amount of samples obtained in a single day is gradually accumulated, the culture pressure is greatly increased, and therefore, the realization of high-throughput culture is also the culture requirement of organoids.

Currently, high throughput culture is commonly used for drug screening in the field of drug development. Cells are usually inoculated into culture wells of a high-throughput culture plate, and different types or different concentrations of drugs are added into different wells, so that pharmacodynamic and pharmacological analysis of different drugs with various concentrations can be realized in a short time. Continuous perfusion requires that each hole or each group of holes is communicated with at least one pipeline for liquid supply and drainage, for example, a culture plate has 96 holes, and if the 96 holes are individually connected with inlet and outlet pipelines and valves for controlling the pipelines, the perfusion system is very large and inconvenient to use.

SUMMERY OF THE UTILITY MODEL

The embodiment of the utility model provides an aim at: provided is a perfusion culture system which is simple in structure.

In order to achieve the purpose, the utility model adopts the following technical proposal:

the utility model provides a perfusion culture system, including cultivateing the board, be provided with a plurality of cultivation holes on the cultivation board, the below of cultivateing the board is provided with the flexible tube, cultivate the hole through first intercommunicating pore with the flexible tube intercommunication, the below of flexible tube is provided with a plurality of shutoff pieces, shutoff piece selectivity is close to and supports tightly the flexible tube, shutoff piece is adjacent to first intercommunicating pore.

As a preferable scheme of the perfusion culture system, the blocking piece comprises a magnetic blocking column, a solenoid valve for controlling the blocking column to move is arranged below the blocking column, the blocking column is connected with the solenoid valve through a sliding rail, and the blocking column can slide in the sliding rail.

As a preferable scheme of the perfusion culture system, an elastic member is arranged between the blocking column and the electromagnetic valve, the elastic member always has a tendency of driving the blocking column to approach and abut against the flexible pipe, and when the electromagnetic valve is powered on, the blocking column is far away from the flexible pipe.

As a preferable scheme of the perfusion culture system, an elastic member is arranged between the blocking column and the electromagnetic valve, the elastic member always has a tendency of driving the blocking column to be away from the flexible pipe, and when the electromagnetic valve is powered on, the blocking column is close to and abuts against the flexible pipe.

As an optimal scheme of the perfusion culture system, the blocking piece comprises a screw, a locking block and a limiting plate, the side face of the locking block is abutted to the limiting plate, the locking block is in threaded connection with the screw, the screw is connected with a motor, and the motor drives the screw to rotate so as to drive the locking block to move along the axis direction of the screw.

As a preferable scheme of the perfusion culture system, the plugging member comprises an air blowing device, the air blowing device blows air towards the flexible tube to form an air column, and the air column abuts against the flexible tube.

As a preferable scheme of the perfusion culture system, a groove is concavely arranged on the lower surface of the culture plate, a flexible film is attached to the lower surface of the culture plate, the groove and the flexible film form the flexible tube, and the groove is communicated with the culture hole through the first communication hole.

As a preferable scheme of the perfusion culture system, a plurality of second communication holes are arranged on the tube wall of the flexible tube, and one second communication hole is communicated with at least one first communication hole.

The perfusion culture system preferably includes a mounting plate disposed below the culture plate, and the plurality of plugging members are mounted on the mounting plate.

As a preferable scheme of the perfusion culture system, one of the mounting plate and the culture plate is provided with a first positioning part, and the other is provided with a second positioning part, and the first positioning part and the second positioning part are matched for positioning.

As a preferable scheme of the perfusion culture system, a diaphragm is arranged between the first communication hole and the flexible pipe, and selectively traps culture and liquid.

The utility model has the advantages that: the blocking piece is arranged below the flexible pipe and located outside the flexible pipe, so that a pipeline device and a control device of the perfusion culture system can be separated, the flexible pipe and the blocking piece respectively belong to the pipeline device and the control device, the structure of the perfusion culture system is simplified, and the operation difficulty of high-flux perfusion culture is reduced; through setting up the shutoff piece and being close to first through-hole, when the shutoff piece is close to and supports tight flexible pipe, under the extrusion of the lower surface of culture plate and shutoff piece terminal surface, the flexible pipe is compressed tightly, and the shutoff effect has been realized to the shutoff piece.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and examples.

FIG. 1 is a schematic view of a perfusion culture system (flexible tube circulation) according to an embodiment of the present invention.

Fig. 2 is a schematic view of a perfusion culture system (flexible tube plugging) according to an embodiment of the present invention.

FIG. 3 is a partial schematic view of a plate and flexible tube according to an embodiment of the present invention.

In the figure:

1. culturing the plate; 101. a culture well; 102. a first communication hole; 103. a positioning column; 2. a flexible tube; 3. a blocking member; 4. mounting a plate; 5. a diaphragm.

Detailed Description

In order to make the technical problems, technical solutions and technical effects achieved by the present invention more clear, the embodiments of the present invention will be described in further detail with reference to the accompanying drawings, and obviously, the described embodiments are only some embodiments, not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without creative efforts belong to the protection scope of the present invention.

As shown in FIG. 1, the utility model provides a pair of perfusion culture system, including culture plate 1, be provided with a plurality of cultivation holes 101 on culture plate 1, the below of culture plate 1 is provided with flexible pipe 2, cultivates hole 101 and communicates through first communicating hole 102 and flexible pipe 2, and the below of flexible pipe 2 is provided with a plurality of shutoff pieces 3, and shutoff piece 3 is adjacent to first communicating hole 102, when needing to close flexible pipe 2, can be close to and support tight flexible pipe 2 with shutoff piece 3. Referring to fig. 2, the flexible tube 2 is squeezed and blocked by the blocking member 3, and the liquid in the flexible tube 2 does not flow. The blocking piece 3 is arranged below the flexible pipe 2, the blocking piece 3 is positioned outside the flexible pipe 2, a pipeline device and a control device of the perfusion culture system can be separated, and the flexible pipe 2 and the blocking piece 3 respectively belong to the pipeline device and the control device, so that the structure of the perfusion culture system is simplified, and the operation difficulty of high-flux perfusion culture is reduced; through setting up shutoff piece 3 and being close to first through-hole 102, when shutoff piece 3 was close to and supports tight flexible tube 2, under the extrusion of the lower surface of culture plate 1 and shutoff piece 3 terminal surface, flexible tube 2 was compressed tightly, and shutoff piece 3 has realized the shutoff effect.

Of course, to ensure the blocking function of the blocking piece 3, the blocking piece 3 is disposed at the upstream side of the first communication hole 102, and when the blocking piece 3 approaches and tightly abuts against the flexible pipe 2, the liquid in the flexible pipe 2 cannot flow into the first communication hole 102 located at the downstream.

In this embodiment, two sets of culture wells 101 are disposed on a culture plate 1 as an example, each culture plate 1 is disposed with 96 culture wells 101, each culture well 101 is communicated with a flexible tube 2 through a first communication hole 102, the 96 culture wells 101 are divided into a culture group a and a culture group B, and a plugging member 3 is disposed between the culture well 101 and the flexible tube 2 in each set, when in use, if the culture group a needs to be perfused independently, the plugging member 3 of the culture group B can be close to and abutted against the flexible tube 2, at this time, the flexible tube 2 of the culture group B is plugged, the liquid introduced into the flexible tube 2 can only enter the culture wells 101 of the culture group a, when the liquid is required to be introduced into the culture wells 101 of the culture group a and the culture group B simultaneously, the plugging member 3 of the culture group B can be moved away from the flexible tube 2, so as to realize the communication of the flexible tubes 2 of the culture group B, at this time, the liquid introduced into the flexible tube 2 can enter the culture well 101 of the culture group A and the culture group B at the same time.

Referring to fig. 3, the diameter of the first communicating hole 102 is much smaller than that of the culture hole 101, and the culture can be inoculated at any position inside the culture hole 101 avoiding the first communicating hole 102; of course, the diaphragm 5 can be arranged between the first communicating hole 102 and the flexible tube 2, so that the culture in the culture hole 101 can be prevented from falling into the flexible tube 2, and the normal operation of the culture operation can be ensured.

The membrane 5 is used for intercepting a culture (macromolecular structure) and preventing the culture from entering the flexible tube 2, the membrane 5 can be a porous membrane and a porous material, the porous membrane comprises a small-aperture membrane and a large-aperture membrane, wherein the aperture range of the small-aperture membrane is 1 nanometer-10 micrometers, the membranes with different apertures have different filtering functions, the membrane with the smallest aperture (1 nanometer-50 nanometers) is used for filtering and exchanging small molecules, the membrane with the slightly larger aperture (10 nanometers-100 nanometers) is used for filtering and collecting cell-producing products such as antibodies and the like, and the membrane with the larger aperture (50 nanometers-10 micrometers) is used for collecting cells and exosomes; the material of the large-aperture membrane partition layer is generally a porous material, the aperture size is 0.1 micrometer to 10 millimeters, wherein the partition material of 0.1 micrometer to 5 micrometers is used for isolating cells from a culture solution, and the cells are ensured not to penetrate through a membrane to enter the perfusion culture solution in the perfusion culture process. The 5-micrometer-10-millimeter segmentation material is used for isolating 3D cell clusters (the size of gel drops formed by wrapping organoids and cell clusters in gel is generally 1-10 millimeters) or isolated tissues, organs and perfusion culture layers. The diaphragm 5 may be a single-layer thin film (generally with a pore size of 0.1 micron to 100 microns), a single-layer thick film (generally with a pore size of 100 microns to 10 millimeters), or a multi-layer film structure with a thin film thickening film as a support film, and the support film may be above the thin film or below the thin film. Of course, the membrane 5 may also be a semi-permeable membrane, and in particular, the semi-permeable membrane arranged between the first communication hole 102 and the flexible tube 2 is a membrane allowing only certain molecules and ions to diffuse in and out, and generally speaking, the semi-permeable membrane allows only ions and small molecular substances to pass through, while bio-macromolecular substances cannot freely pass through the semi-permeable membrane, because the pores of the semi-permeable membrane are larger than the ions and small molecular substances, but smaller than the bio-macromolecules (e.g. proteins, starches, etc.). Parchment, cellophane, etc. belong to semi-permeable membranes, for example, cellophane only allows water to permeate sucrose solution, but sucrose molecules cannot permeate; the animal's bladder allows water to permeate through it but not alcohol molecules; the hot palladium or platinum is permeable to hydrogen, but impermeable to argon molecules. The semipermeable membrane can be made of various high molecular materials and is used for separating substances with different molecular weights, fixing osmotic pressure, gas partial pressure and the like. The semipermeable membrane is mainly used for reverse osmosis and ultrafiltration in membrane separation technology. When the reverse osmosis process is applied, the membrane is called a reverse osmosis membrane, and is a thin film with water-based groups, and the membrane not only has a screening function, but also has a preferential adsorption function on water molecules. The semipermeable membrane may be in the form of a plate, a tube, or a hollow fiber.

In one embodiment, the blocking piece 3 comprises a magnetic blocking column, an electromagnetic valve for controlling the movement of the blocking column is arranged below the blocking column, the blocking column and the electromagnetic valve are connected through a sliding rail, and the blocking column can slide in the sliding rail. Through setting up the solenoid valve and having magnetic shutoff piece 3, the solenoid valve can utilize the magnetism control shutoff piece 3 of shutoff piece 3 to be close to or keep away from flexible pipe 2, and the slide rail can provide the guide effect for the removal of shutoff post, avoids shutoff piece 3 to appear the offset when removing, has guaranteed the shutoff effect of shutoff piece 3 to flexible pipe 2.

Specifically, an elastic piece is arranged between the blocking column and the electromagnetic valve, in one embodiment, the elastic piece always has a tendency of driving the blocking column to approach and abut against the flexible pipe 2, when the electromagnetic valve is powered on, the blocking column is far away from the flexible pipe 2 under the magnetic action, at the moment, the flexible pipe 2 circulates, and liquid can flow into the first communication hole 102, in this embodiment, the electromagnetic valve is powered on, the flexible pipe 2 circulates, the electromagnetic valve is powered off, and the flexible pipe 2 blocks, so that the blocking column is suitable for the normally closed flexible pipe 2; in another embodiment, the elastic member always has a tendency of driving the blocking member 3 to be away from the flexible tube 2, when the electromagnetic valve is powered on, the blocking column is close to and abuts against the flexible tube 2 under the magnetic effect, at the moment, the flexible tube 2 is squeezed to block, liquid in the flexible tube 2 does not flow, in this embodiment, the electromagnetic valve is powered on, the flexible tube 2 blocks, the electromagnetic valve is powered off, and the flexible tube 2 circulates, so that the blocking member 3 is suitable for the normally open flexible tube 2. The plugging columns with different control modes are arranged, and the types of the plugging columns can be selected according to the culture requirements, so that the cost in the culture process is reduced. Of course, the magnetic force generated by the electromagnetic valve can be controlled by controlling the current, so as to control the moving distance of the plugging column, when the flow rate of the liquid in the flexible tube 2 needs to be controlled, the plugging column can be close to the flexible tube 2, so that the tube wall of the flexible tube 2 is slightly deformed, and at the moment, the flexible tube 2 is still in a circulating state.

In an embodiment, shutoff piece 3 can be the device including the screw rod, locking block and limiting plate, the side and the limiting plate butt of locking block, locking block and screw rod threaded connection, screw rod and motor are connected, because locking block and screw rod threaded connection, when the motor drives the screw rod and rotates, the locking block has follows screw rod pivoted trend, and the side and the limiting plate butt of locking block, under the effect of limiting plate, the locking block can't take place to rotate, the motion of locking block changes the axis direction removal along the screw rod into, when motor corotation, the locking block is close to flexible pipe 2 and removes and supports tight shutoff with flexible pipe 2, when the motor reversal, flexible pipe 2 is kept away from to the locking block, flexible pipe 2 circulates. The setting of screw rod and motor can the displacement of accurate control locking block, when the velocity of flow of liquid in the flexible pipe 2 of needs control, can be close to flexible pipe 2 with the locking block, makes the slight deformation appear in the pipe wall of flexible pipe 2, and at this moment, still is in the state of circulation in the flexible pipe 2.

In other embodiments, the blocking member 3 may be an air blowing device, specifically, an air outlet is provided on the air blowing device, the air outlet faces the flexible tube 2, the air blowing device blows air towards the flexible tube 2 to form an air column, air pressure generated by the air column presses and deforms the tube wall of the flexible tube 2, and at this time, the flexible tube 2 is blocked. Of course, the air flow speed blown out by the air blowing device can be controlled to control the deformation of the tube wall of the flexible tube 2, so as to control the flow speed of the liquid in the flexible tube 2.

Specifically, in an embodiment, the flexible tube 2 is an independent tube, the tube wall of the flexible tube 2 is provided with a plurality of second communicating holes, and according to the requirement of culture, one second communicating hole can be communicated with one first communicating hole 102, so that the culture solution in each culture hole 101 can be independently controlled, or one second communicating hole can be communicated with a plurality of first communicating holes 102, so that the culture solution in the culture holes 101 can be controlled in batch, and in order to ensure the differentiation and maturation of the culture in the culture position, the diaphragm 5 is arranged between the second communicating hole and the first communicating hole 102.

Specifically, in this embodiment, a concave groove is formed on the lower surface of the culture plate 1, a flexible film is attached to the lower surface of the culture plate 1, the groove and the flexible film together form the flexible tube 2, the groove is communicated with the culture hole 101 through the first communication hole 102, and the diaphragm 5 is arranged between the groove and the first communication hole 102. The flexible tube 2 is a combination of a groove on the lower surface of the culture plate 1 and a flexible membrane, so that the positioning difficulty between the flexible tube 2 and the first communication hole 102 can be reduced, and the processing cost of the perfusion culture system is reduced.

Specifically, the perfusion culture system further comprises a mounting plate 4, wherein the mounting plate 4 is arranged below the culture plate 1, and a plurality of plugging pieces 3 are mounted on the mounting plate 4. In the high-throughput culture mode, the number of culture wells 101 on a single culture plate 1 is large (e.g., 96 wells, 384 wells, 1536 wells), and therefore, a plurality of blocking members 3 need to be arranged to control the circulation and the closure of the flexible tube 2, and by arranging the mounting plate 4, the plurality of blocking members 3 can be integrated into a control module, thereby reducing the difficulty in mounting the perfusion culture system.

Specifically, the culture plate 1 is provided with a first positioning member, and the mounting plate 4 is provided with a second positioning member, in this embodiment, the first positioning member is a positioning column 103, and the second positioning member is a positioning hole, so that the positioning column 103 can be inserted into the positioning hole during mounting, and of course, the positioning hole can also be provided on the culture plate 1, and the positioning column 103 can be provided on the mounting plate 4; because the plugging piece 3 needs to be respectively aligned with the flexible pipe 2 and is provided with the positioning hole and the positioning column 103, the mounting and positioning difficulty of the device can be reduced, and the assembly efficiency of the perfusion culture system is improved; of course, in other embodiments, other positioning devices such as magnets, snaps, etc. may be used to position the mounting plate 4 and the growth plate 1.

Specifically, when shutoff piece 3 is the device including screw rod, locking piece and limiting plate, mounting panel 4 can act as the limiting plate, and at this moment, need set up the through-hole on mounting panel 4, with the side of locking piece and the inner wall butt of through-hole, under the effect of through-hole, the locking piece can be followed the axis direction removal of through-hole.

Claims

1. The perfusion culture system is characterized by comprising a culture plate, wherein a plurality of culture holes are formed in the culture plate, a flexible pipe is arranged below the culture plate, the culture holes are communicated with the flexible pipe through first communication holes, a plurality of plugging pieces are arranged below the flexible pipe, the plugging pieces are selectively close to and abut against the flexible pipe, and the plugging pieces are adjacent to the first communication holes.

2. The perfusion culture system of claim 1, wherein the blocking piece comprises a magnetic blocking column, a solenoid valve for controlling the blocking column to move is arranged below the blocking column, the blocking column and the solenoid valve are connected through a sliding rail, and the blocking column can slide in the sliding rail.

3. The perfusion culture system of claim 2, wherein an elastic member is disposed between the blocking column and the solenoid valve, the elastic member always has a tendency to drive the blocking column to approach and abut against the flexible tube, and the blocking column is away from the flexible tube when the solenoid valve is energized.

4. The perfusion culture system of claim 2, wherein an elastic member is disposed between the blocking column and the solenoid valve, the elastic member always has a tendency to drive the blocking column away from the flexible tube, and when the solenoid valve is energized, the blocking column approaches and abuts against the flexible tube.

5. The perfusion culture system of claim 1, wherein the blocking piece comprises a screw, a locking block and a limiting plate, the side surface of the locking block abuts against the limiting plate, the locking block is in threaded connection with the screw, the screw is connected with a motor, and the motor drives the screw to rotate to drive the locking block to move along the axial direction of the screw.

6. The perfusion culture system of claim 1, wherein the block includes an insufflation device that insufflates towards the flexible tube forming a column of air that abuts against the flexible tube.

7. The perfusion culture system of claim 1, wherein a groove is concavely arranged on the lower surface of the culture plate, a flexible membrane is attached to the lower surface of the culture plate, the groove and the flexible membrane form the flexible tube, and the groove is communicated with the culture hole through the first communication hole.

8. The perfusion culture system according to claim 1, wherein a plurality of second communication holes are arranged on the tube wall of the flexible tube, and one of the second communication holes is communicated with at least one of the first communication holes.

9. The perfusion culture system of claim 1, comprising a mounting plate disposed below the culture plate, the plurality of plugs being mounted on the mounting plate.

10. The perfusion culture system of claim 9, wherein one of the mounting plate and the culture plate is provided with a first positioning element and the other is provided with a second positioning element, and the first positioning element and the second positioning element are cooperatively positioned.

11. The perfusion culture system of claim 1, wherein a membrane is disposed between the first communication hole and the flexible tube, the membrane selectively entrapping culture and liquid.