A porous scaffold perfusion device for inoculating cells

Abstract

A porous scaffold perfusion device for inoculating cells comprises a base, wherein the base has two columns and three rows of six disc grooves and square grooves for supporting a perfusion chamber, wherein the perfusion chamber consists of an inoculation chamber and a perfusion cover, wherein the inoculation chamber can be composed of a culture dish and a six-well plate, which are common general cell culture consumables in laboratories, and the perfusion cover is prepared by 3D printing casting molding technology, wherein a groove is provided at the center of the cavity for fixing the porous scaffold material, and the unique physical cavity can make the fluid shear the seed cells in the porous scaffold to the greatest extent and stimulate their growth, and the top of the perfusion cover has two holes for inlet and outlet of the fluid, which are connected to a rubber tube through a change-of-direction joint, and the perfusion chamber can be connected to other parallel perfusion chambers through the two ends of a three-way joint to form a maximum of six synchronous parallel experiments, and can also form synchronous experiments of experimental groups and control groups, and the device has a clever structure, simple operation, strong versatility, and controllable flow rate, which greatly improves the experimental efficiency.

Description

Technical Field

[0001] The invention belongs to the field of cell mechanics and biology, and in particular relates to a porous support perfusion device for inoculating cells. Background Art

[0002] In clinical treatment, there are many diseases that cannot be treated with ordinary chemical drugs and physical therapy. For example, leukemia patients need bone marrow transplantation to be fundamentally treated, and they need to use stem cell culture to produce bone marrow hematopoietic stem cells. For example, bone cartilage tissue caused by trauma or degenerative diseases is difficult to regenerate, and there are still many disadvantages in the effective solutions.

[0003] The proposal, establishment and development of tissue engineering not only provides a reliable method for repairing, replacing or regenerating defective tissues. As a site for cell transplantation and a template for tissue regeneration, tissue engineering scaffolds have unique advantages. They can not only simulate the microenvironment of cell growth inside the human body, but also have a wide range of applications and can have a good effect on constructing structural tissues such as bone, cartilage and skin.

[0004] Bioscaffolds can be used for three-dimensional cell culture, providing a good living environment for them. The stimulation of cells in bioreactors is a key area of ​​research in the fields of biomechanics and regenerative medicine. It can create a three-dimensional space similar to the in vivo environment to proliferate high-quality cells that are difficult to culture in culture systems. The organic combination of cell tissue engineering and in vitro perfusion culture technology is a key technology for in vitro culture of tissue cells.

[0005] Currently, most in vitro perfusion devices are mainly used for culturing cells, and there is little description of porous scaffold perfusion devices inoculated with cells; the devices are designed based on their own experimental needs and only solve their specific problems, and are not suitable for a large number of studies based on cell mechanics; the perfusion parts of the devices are of different sizes, have many components, and are cumbersome to assemble, which places high demands on the experimental environment. Summary of the invention

[0006] The present invention discloses a porous scaffold perfusion device for inoculating cells, which improves and upgrades the traditional in vitro perfusion device. Based on common laboratory common cell culture consumables culture dishes and six-well plates, combined with the specific shape of biological scaffold materials, a reasonable mold is designed by reverse engineering, and a sealing cover with good sealing performance is prepared by 3D printing casting molding technology. The in vitro perfusion device for cell tissue engineering is built with the aid of a peristaltic drive, a rubber tube, a liquid storage bottle, a three-way joint, a change-of-direction joint, etc. The bottom adopts a unique flow field slope, which is specially used for perfusion culture of seed cells in porous scaffolds to achieve the purpose of cell stimulation growth. The device has the characteristics of simple operation, convenient installation and disassembly, personalized matching, and strong practicality.

[0007] The technical solution adopted by the present invention to solve the technical problem is: a porous scaffold perfusion device for inoculating cells, comprising a base, wherein the base has two columns and three rows of six disc grooves for supporting a perfusion chamber, the perfusion chamber is composed of an inoculation chamber and a perfusion cover, the inoculation chamber can be composed of a common laboratory common cell culture consumables culture dish and a six-well plate, the perfusion cover is prepared by 3D printing casting molding technology, the center of the cavity is provided with a groove, the porous scaffold material can be fixed, the unique flow field slope of the bottom surface enables the flow night to shear the seed cells in the porous scaffold to the maximum extent and stimulate their growth, and the top of the perfusion cover is provided with two inlet and outlet holes, the two holes are connected to the rubber tube through a change-of-direction joint, the rubber tube is connected to the same row of perfusion chambers through the two ends of a three-way joint, and the non-same row of perfusion chambers are connected in parallel through the three-way joint, so that up to six synchronous parallel experiments can be formed, and synchronous experiments of experimental groups and control groups can also be formed, the base has a threaded hole, and the perfusion chamber is fixed with a positioning plate; the device has a clever structure, simple operation, strong versatility, controllable flow rate, and greatly improves the experimental efficiency.

[0008] The beneficial effects of the present invention are as follows: the present invention has a novel structure and is easy to operate; the inoculation chamber can be constructed by culture dishes and six-well plates, which are common cell culture consumables in laboratories, so that the core components of the device have a uniform size; the perfusion cover is made of addition-type silicone material, which reduces the components of the device while ensuring the sealing of the perfusion chamber; the unique physical structure of the bottom can produce a more ideal flow field, thereby realizing the physical environment required by the cells; the three-way connector can realize synchronous perfusion of each perfusion chamber, and a large number of studies based on porous scaffolds inoculated with cells can be carried out. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0010] Figure 2 For the present invention Figure 1 Sectional view of AA.

[0011] Figure 3 For the present invention Figure 1 Enlarged cross-sectional view of the perfusion chamber assembly.

[0012] Figure 4 For the present invention Figure 1 Magnified top view of the mid-perfusion cap. DETAILED DESCRIPTION

[0013] A porous support perfusion device for inoculating cells, comprising a base 1, characterized in that two columns and three rows of six disc grooves are provided on both sides of the upper surface of the base 1, which can support a perfusion chamber of the culture dish type, a square groove is provided in the middle, and slots are provided at the four corners, which can be used to support a perfusion chamber of the six-well plate type, the perfusion chamber is composed of an inoculation chamber and a perfusion cover, the inoculation chamber can be constructed by a culture dish and a six-well plate, which are common cell culture consumables in laboratories, a liquid inlet of the perfusion cover 41 is inserted with a change-direction joint 95, and the outlet end is connected to a rubber tube 84, a liquid inlet of the perfusion cover 43 is inserted with a change-direction joint 97, and the outlet end is connected to a rubber tube 85, the rubber tube 84 and the rubber tube 85 are connected through the two ends of a three-way joint 101, a liquid inlet of the perfusion cover 42 is inserted with a change-direction joint 91, and the outlet end is connected to a rubber tube 86, a liquid inlet of the perfusion cover 44 is inserted with a change-direction joint 93, and the outlet end is connected to the rubber tube 87 The rubber tube 86 and the rubber tube 87 are connected through the two ends of the three-way joint 102, the other end of the three-way joint 101 is connected to the rubber tube 82, the other end of the three-way joint 102 is connected to the rubber tube 83, the rubber tube 82 and the rubber tube 83 are connected through the two ends of the three-way joint 100, the other end of the three-way joint 100 is connected to one end of the main rubber tube 81, the other end of the main rubber tube 81 is connected to the inlet of the liquid storage bottle 7, and the tube wall of the main rubber tube 81 is inserted into the peristaltic drive 6.

[0014] The liquid outlet of the perfusion cover 41 is inserted with a direction-changing joint 96, which is connected to one end of the three-way joint 102 through a rubber ring 110; the liquid outlet of the perfusion cover 42 is inserted with a direction-changing joint 92, which is connected to one end of the three-way joint 102 through a rubber tube 88; the liquid outlet of the perfusion cover 43 is inserted with a direction-changing joint 98, which is connected to one end of the three-way joint 103 through a rubber ring 111; the liquid outlet of the perfusion cover 44 is inserted with a direction-changing joint 94, which is connected to one end of the three-way joint 103 through a rubber tube 89.

[0015] The three-way joint 102 is connected to one end of the rubber tube 90, the three-way joint 103 is connected to one end of the rubber tube 91, the other end of the rubber tube 90 and the other end of the rubber tube 91 are connected through the three-way joint 104, and the other end of the three-way joint 104 is connected to the outlet of the liquid storage bottle 7 through the rubber tube 92.

[0016] The perfusion cap 41 is made of addition type silicone material and is prepared by 3D printing molding and casting technology, specifically: a reverse engineering method is used to design a reasonable mold according to the physical size of the biological scaffold, and the addition type silicone stock solution casting method is used for molding.

[0017] The top of the perfusion cover 41 has a liquid inlet and a liquid outlet, and a groove in the middle of the bottom for fixing the biological scaffold. The bottom has a unique flow field slope, which allows the flow to fully and evenly shear the seed cells in the porous scaffold and stimulate their growth. The inlet wall is a tapered wall, which has the advantage of producing a more ideal flow field and having the function of buffering liquid; the outlet wall is a gradually expanding wall, which has the advantage of buffering the liquid outflow.

[0018] During operation, the porous scaffold material 8 inoculated with cells is placed in the inoculation chamber, the perfusion cover is inserted therein, the positioning plate 21 and the positioning plate 22 are used to press the perfusion chamber, the liquid storage bottle 7 is filled with cell culture fluid, the peristaltic drive 6 provides a power source, and the cell culture fluid is pumped out from the inlet of the liquid storage bottle 7 and flows through the main rubber tube 81, which is connected to one end of the three-way connector 100 to perform fluid diversion. The three-way connector 100 The other two ends are connected to one end of the rubber tube 82 and the rubber tube 83 respectively, and the diverted liquid flows through the rubber tube 82 and the rubber tube 83 respectively. The other end of the rubber tube 82 is connected to one end of the three-way joint 101, and the other end of the rubber tube 83 is connected to one end of the three-way joint 102. The liquid is secondary diverted through the three-way joint 101 and the three-way joint 102. A branch of the diverted liquid flows to the entrance of the perfusion chamber 41 and the perfusion chamber 43 respectively through the rubber tube 84 and the rubber tube 85 connected to the other two ends of the three-way joint 101. The entrances of the perfusion chamber 41 and the perfusion chamber 43 are respectively inserted with a changing joint 95 and a changing joint 98, so that the liquid flows into the perfusion chamber and flows through the porous scaffold inoculated with cells to stimulate cell growth. Similarly, the other branch of the flow after diversion flows to the entrances of the perfusion chamber 42 and the perfusion chamber 44 through the rubber tube 86 and the rubber tube 87 connected to the other two ends of the three-way joint 102, respectively. The entrances of the perfusion chamber 42 and the perfusion chamber 44 are inserted with the reversing joint 91 and the reversing joint 94, so that the flow flows into the perfusion chamber and flows through the porous scaffold inoculated with cells to stimulate cell growth. One branch of the flow converges from the three-way joint 102 and the rubber tube 88, and the other branch converges from the three-way joint 103 and the rubber tube 89. The converged flow flows through the rubber tube 90 and the rubber tube 91, and converges again at the three-way joint 104, and finally flows into the liquid storage bottle 7 through the rubber tube 92. The above is a cycle process.

[0019] What is described here is the synchronous perfusion of four perfusion chambers, and the other two perfusion chambers form the experimental control group. It is also possible to perform synchronous perfusion of six perfusion chambers, and the method and principle are the same as above.

[0020] The inoculation chamber described here uses a common laboratory consumable culture dish, and a common laboratory consumable six-well plate can also be used. The method and principle are the same as above.

[0021] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes, and they can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the illustrations shown and described herein.

Claims

1. A porous scaffold perfusion device for inoculating cells, characterized in that: The base has two columns and three rows of six disc grooves and square grooves for supporting the perfusion chamber. The perfusion chamber is composed of an inoculation chamber and a perfusion cover. The inoculation chamber can be composed of a common laboratory cell culture consumables culture dish and a six-well plate. The perfusion cover is prepared by 3D printing casting technology. There is a groove in the center of the cavity to fix the porous scaffold material. The unique physical cavity can make the fluid shear the seed cells in the porous scaffold to the greatest extent and stimulate their growth. There are two holes for inlet and outlet of the perfusion cover at the top. The two holes are connected to the rubber tube through a change-of-direction joint. The perfusion chamber can be connected to other parallel perfusion chambers through the two ends of the three-way joint to form up to six synchronous parallel experiments, and can also form synchronous experiments of experimental and control groups. The porous support perfusion device for inoculating cells is characterized in that two columns and three rows of six disc grooves are arranged on both sides of the upper surface of the base (1), and a square groove is arranged in the middle, which is used to support the perfusion chamber. The perfusion chamber is composed of an inoculation chamber and a perfusion cover. The liquid inlet of the perfusion cover (41) is inserted with a change-direction joint (95), and the outlet end is connected to the rubber tube (84). The liquid inlet of the perfusion cover (43) is inserted with a change-direction joint (97), and the outlet end is connected to the rubber tube (85). The rubber tube (84) and the rubber tube (85) are connected at both ends through a three-way joint (101). The liquid inlet of the perfusion cover (42) is inserted with a change-direction joint (91), and the outlet end is connected to the rubber tube (84). The rubber tube (86) is connected to the liquid inlet of the perfusion cover (44) with a change-direction joint (93), the outlet end is connected to the rubber tube (87), the rubber tube (86) and the rubber tube (87) are connected through the two ends of a three-way joint (102), the other end of the three-way joint (101) is connected to the rubber tube (82), the other end of the three-way joint (102) is connected to the rubber tube (83), the rubber tube (82) and the rubber tube (83) are connected through the two ends of a three-way joint (100), the other end of the three-way joint (100) is connected to one end of the main rubber tube (81), and the other end of the main rubber tube (81) is connected to the inlet of the liquid storage bottle (7); The liquid outlet of the perfusion cover (41) is inserted with a direction-changing joint (96), and the direction-changing joint (96) is connected to one end of the three-way joint (102) via a rubber ring (110); the liquid outlet of the perfusion cover (42) is inserted with a direction-changing joint (92), and the direction-changing joint (92) is connected to one end of the three-way joint (102) via a rubber tube (88); the liquid outlet of the perfusion cover (43) is inserted with a direction-changing joint (98), and the direction-changing joint (98) is connected to one end of the three-way joint (103) via a rubber ring (111); the liquid outlet of the perfusion cover (44) is inserted with a direction-changing joint (94), and the direction-changing joint (94) is connected to one end of the three-way joint (103) via a rubber tube (89); The three-way joint (102) is connected to one end of the rubber tube (90), the three-way joint (103) is connected to one end of the rubber tube (91), the other end of the rubber tube (90) and the other end of the rubber tube (91) are connected via a three-way joint (104), and the other end of the three-way joint (104) is connected to the outlet of the liquid storage bottle (7) via the rubber tube (92); The porous support perfusion device for inoculating cells is characterized in that: the upper surface of the base (1) has two columns and three rows of six disc grooves, which can support a culture dish type inoculation chamber, the middle of the upper surface of the base has a square groove, and the four corners have slots, which can be used to support a six-well plate type inoculation chamber, and can realize up to six synchronous parallel experiments, and can also form experimental group and control group synchronous experiments, wherein the perfusion cover (45) and the perfusion cover (46) are control group perfusion covers; The porous scaffold perfusion device for inoculating cells is characterized in that: the perfusion cover (41) is made of addition-type silicone material and can be prepared by 3D printing casting technology, and its material properties ensure the sealing of the perfusion chamber; The porous scaffold perfusion device for inoculating cells is characterized in that: the perfusion cover (41) can form different central cavities according to the shapes of different scaffolds: square, columnar, pyramidal and asymmetric structure; The porous scaffold perfusion device for inoculating cells is characterized in that: the middle of the cavity (41) of the perfusion cover has a groove for fixing the porous scaffold material, the bottom surface has a unique flow field slope, the cavity inlet wall is a tapered type, which changes the flow field of the fluid and has the function of buffering the flow, and the cavity outlet wall is a gradually expanding type, which is conducive to the buffering outflow of the flow; The porous scaffold perfusion device for inoculating cells is characterized in that: the direction-changing joint (96) is inserted into the liquid inlet and liquid outlet of the perfusion cover, one end of which leads to the cavity of the perfusion cover and the other end is connected to the rubber tube; The porous scaffold perfusion device for inoculating cells is characterized in that one end of the three-way connector (91) is connected to the main rubber tube, and the other two ends are connected to the shunt tube for liquid shunting, so that the liquid flows evenly into the perfusion chamber; The porous support perfusion device for inoculating cells is characterized in that the liquid storage bottle (7) has inlet and outlet holes at the bottle mouth.

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