Incubator for driving magnetic nanoparticles to move in tissue cells

Abstract

The invention discloses a culture box for driving magnetic nanoparticles to move in tissue cells, and relates to the field of biological culture.The culture box comprises a box body and a box door, the box body is rotationally connected with the box door, two partition plates are arranged on the lower portion in the box body, a culture chamber is formed between the partition plate on the upper layer and a top plate of the box body, and the partition plate on the lower layer and the top plate of the box body are arranged in the culture chamber. A driving chamber is formed between the two partition plates, and a heating chamber is formed between the partition plate on the lower layer and a bottom plate of the box body; a magnetic driving device is arranged in the culture chamber, a power mechanism is arranged in the driving chamber, and the magnetic driving device is in power connection with the power mechanism; the heating chamber is provided with a heating device, and the heating device is communicated with the culture chamber through a pipeline. The magnetic driving device is arranged in the box body, the magnetic driving device can make nano magnetic particles move under the traction of a magnetic field, and two functions of magnetic driving and cell culture are considered.

Description

Technical Field

[0001] The invention relates to the field of biological culture, and in particular to a culture box for driving magnetic nanoparticles to move in tissue cells. Background Art

[0002] Physiological barriers (pancreatic matrix barrier, blood-brain barrier, blood-cerebrospinal fluid barrier, skin barrier, placental barrier, blood-urine barrier, air-blood barrier, filtration barrier, blood-thymus barrier, etc.) hinder the delivery of drugs. In order to solve such problems, in the 1970s, Widder et al. proposed the concept of magnetically controlled targeted delivery system and conducted research on drug-loaded magnetic microparticles. By driving drug carriers by magnetic field, the precise delivery of anti-tumor drugs can be achieved with minimal impact on organic life. Since then, magnetically controlled targeted therapy has been widely studied. For example, Dreyfus et al. prepared a micron-scale magnetically driven worm. Under the action of a vertical bidirectional magnetic field, the worm was controlled to swing its tail and move in a directional manner in the liquid. However, as the research progressed, researchers needed to observe the movement ability of the above nanoparticles under cells and tissues. At present, common cell culture equipment does not have the function of generating and controlling magnetic fields, and cannot take into account both magnetic drive and cell culture functions. A culture box that meets both functions is needed to solve this problem. Summary of the invention

[0003] The purpose of the present invention is to provide an incubator for driving magnetic nanoparticles to move in tissue cells, so as to solve the problem mentioned in the background technology that the currently common cell culture equipment does not have the magnetic field generation and control functions and cannot take into account both the magnetic drive and cell culture functions.

[0004] The present invention adopts the following technical solution:

[0005] The present invention discloses an incubator for driving magnetic nanoparticles to move in tissue cells, comprising a box body and a door, wherein the box body is rotatably connected to the door, two partitions are arranged at the lower part of the box body, a culture chamber is formed between the upper partition and the top plate of the box body, a driving chamber is formed between the two partitions, and a heating chamber is formed between the lower partition and the bottom plate of the box body.

[0006] A magnetic driving device is arranged in the culture chamber, a power mechanism is arranged in the driving chamber, and the magnetic driving device is connected to the power mechanism in power.

[0007] The heating chamber is provided with a heating device, and the heating device is communicated with the culture chamber through a pipeline.

[0008] Furthermore, the magnetic drive device includes a fixed cross bar, both ends of which are fixedly connected to the inner wall of the box body, a base is fixedly connected to the fixed cross bar, a square frame is arranged on the periphery of the base, the square frame is rotatably connected to the fixed cross bar, grooves are arranged on the inner sides of two frame frames parallel to the fixed cross bar, magnets are arranged in the grooves, and the two grooves face one side of the base; the square frame is dynamically connected to the power mechanism; a detachable annular magnet is sleeved on the fixed cross bar, and the annular magnet is located between the base and the square frame.

[0009] Furthermore, the square frame is rotatably connected to the fixed cross bar via a bearing.

[0010] Furthermore, the power mechanism includes a servo motor, which is arranged on the partition located on the lower side. A driving sprocket is fixedly connected to the output shaft of the servo motor, and the driving sprocket is connected to the driven sprocket through a chain.

[0011] The driven sprocket is rotatably sleeved on the fixed cross bar, and the driven sprocket is fixedly connected to the square frame.

[0012] Furthermore, the heating device includes a heating tube, which is horizontally arranged on the bottom plate of the box. Branch pipes are provided at both ends of the heating tube, and the branch pipes are vertically arranged in the side plates of the box. The top ends of the branch pipes extend to the top of the box and meet. The side walls of the box where the branch pipes are located are provided with heat dissipation holes, an embedded blower and an air heater.

[0013] Furthermore, the heating tubes are arranged in an "S" shape.

[0014] Furthermore, an air inlet and an air outlet are provided on one side panel of the box; a water vapor delivery port is provided on the back panel of the box; a temperature sensor, a humidity sensor, and a gas sensor are provided on the inner side of the top panel of the box; a lamp tube is detachably connected to the inner side of the top panel of the box; and an observation window is provided on the box door.

[0015] Furthermore, a microcomputer and a multi-function display screen are arranged on the upper side of the box; the temperature sensor, the humidity sensor, the gas sensor and the power mechanism are controlled by the microcomputer.

[0016] Compared with the prior art, the beneficial technical effects of the present invention are:

[0017] The present invention provides a magnetic drive device in the box, and the cell culture plate can be placed in the magnetic drive device. The magnetic drive device can make the nano magnetic particles move under the traction of the magnetic field to study the penetration effect of the magnetic field on cell tissue and drug delivery. The present invention has the advantages of taking into account both the functions of magnetic drive and cell culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention will be further described below in conjunction with the accompanying drawings.

[0019] Figure 1 This is a schematic diagram of the main structure of the incubator used to drive the movement of magnetic nanoparticles in tissue cells in Example 1 of the present invention;

[0020] Figure 2 This is a schematic structural diagram of a magnetic drive device, a power mechanism and a heating device of an incubator for driving magnetic nanoparticles to move in tissue cells in Example 1 of the present invention;

[0021] Figure 3 This is a schematic diagram of the block structure of an incubator for driving magnetic nanoparticles to move in tissue cells in Example 1 of the present invention;

[0022] Figure 4 This is a schematic diagram of the arrangement of the heating tubes in the first embodiment of the present invention;

[0023] Figure 5 The results of the cell spheroid observation experiment without an external field strength in Example 2 of the present invention are shown;

[0024] Figure 6 The results of the cell spheroid observation experiment under the laboratory-made external field strength device in Example 2 of the present invention are shown.

[0025] Explanation of the accompanying drawings: 1. Box body; 2. Box door; 3. Partition; 4. Cultivation chamber; 5. Drive chamber; 6. Heating chamber; 7. Magnetic drive device; 7-1. Fixed cross bar; 7-2. Base; 7-3. Frame; 7-4. Groove; 7-5. Ring magnet; 7-6. Bearing; 8. Power mechanism; 8-1. Servo motor; 8-2. Driving sprocket; 8-3. Chain; 8-4. Driven sprocket; 9. Heating device; 9-1. Heating tube; 9-2. Branch pipe; 9-3. Heat dissipation hole; 9-4. Embedded blower; 9-5. Air heater; 10. Air inlet; 11. Air outlet; 12. Water vapor delivery port; 13. Temperature sensor; 14. Humidity sensor; 15. Gas sensor; 16. Lamp tube; 17. Microcomputer; 18. Multi-function display screen; 19. Observation window. DETAILED DESCRIPTION

[0026] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments.

[0027] Embodiment 1

[0028] like Figure 1As shown, in this embodiment, a culture box for driving magnetic nanoparticles to move in tissue cells is disclosed, including a box body 1 and a box door 2, the box body 1 is rotatably connected to the box door 2, two partitions 3 are arranged at the lower part of the box body 1, a culture chamber 4 is formed between the upper partition 3 and the top plate of the box body 1, a driving chamber 5 is formed between the two partitions 3, and a heating chamber 6 is formed between the lower partition 3 and the bottom plate of the box body 1. A magnetic driving device 7 is arranged in the culture chamber 4, a power mechanism 8 is arranged in the driving chamber 5, and the magnetic driving device 7 is connected to the power mechanism 8 in power. A heating device 9 is arranged in the heating chamber 6, and a heat dissipation hole is arranged on the bottom side of the heating chamber 6 for heat dissipation. The heating device 9 is connected to the culture chamber 4 through a pipeline.

[0029] An air inlet 10 and an air outlet 11 are provided on one side plate of the box 1, and different gases can be connected externally according to the culture requirements. A water vapor delivery port 12 is provided on the back plate of the box 1, and the water vapor delivery port 12 can be connected externally with water vapor to control the humidity in the culture chamber 4. A temperature sensor 13, a humidity sensor 14, and a gas sensor 15 are provided on the inner side of the top plate of the box 1, and a lamp tube 16 is detachably connected to the inner side of the top plate of the box 1. A microcomputer 17 and a multifunctional display screen 18 are provided on the upper side of the box 1, and the temperature sensor 13, the humidity sensor 14, the gas sensor 15 and the power mechanism 8 are controlled by the microcomputer 17. The temperature sensor 13, the humidity sensor 14, the gas sensor 15 are connected to the microcomputer 17 through pipelines, and the microcomputer 17 is connected to the multifunctional display screen 18 through pipelines. An observation window 19 is provided on the box door 2. The temperature sensor 13 and the humidity sensor 14 can monitor the temperature and humidity in the box 1 in real time. The humidity detector 14 is used to detect the water content in the incubator to control the input of water vapor, and the gas sensor 15 is used to detect the air components in the incubator to adjust the input speed of external gas.

[0030] like Figure 2 As shown, the magnetic drive device 7 includes a fixed crossbar 7-1, both ends of which are fixedly connected to the inner wall of the box body 1, and a base 7-2 is fixedly connected to the fixed crossbar 7-1. The size of the base 7-2 is adapted to the cell culture plate, and the cell culture plate can be embedded in the base 7-2. A square frame 7-3 is arranged on the periphery of the base 7-2, and the square frame 7-3 is rotatably connected to the fixed crossbar 7-1. The inner sides of the two frames parallel to the fixed crossbar 7-1 of the square frame 7-3 are provided with grooves 7-4, and the two grooves 7-4 face one side of the base 7-2. The grooves 7-4 are used to embed different magnets to provide a radial magnetic field. The square frame 7-3 is connected to the power mechanism 8 in a power connection. Bearings 7-6 are arranged in the two frames connected to the square frame 7-3 and the fixed crossbar 7-1, and the bearings 7-6 are sleeved on the fixed crossbar 7-1. With the help of the bearings 7-6, the square frame 7-3 can realize arbitrary rotation on the fixed crossbar 7-1.

[0031] The fixed crossbar 7-1 is provided with a detachable annular magnet 7-5, which is used to provide an axial magnetic field. The annular magnet 7-5 is located between the base 7-2 and the square frame 7-3.

[0032] In this embodiment, the magnets in the two grooves 7-4 are symmetrically distributed and periodically change the vertical magnetic field during rotation. Therefore, they need to be set as a group of two. The annular magnet 7-5 provides a transverse traction magnetic field to provide horizontal pulling force. Therefore, only one is required in this embodiment.

[0033] In this embodiment, to avoid interference during operation, the fixed crossbar 7-1, the base 7-2, and the frame 7-3 should be made of non-magnetic materials, such as titanium, aluminum, zirconium, etc. And to minimize wear, the frame 7-3 should be made of materials with low density, not easy to wear, and not easy to deform, such as titanium alloy.

[0034] like Figure 2 As shown, the power mechanism 8 includes a servo motor 8-1, which is arranged on the partition 3 located at the lower side. A driving sprocket 8-2 is fixedly connected to the output shaft of the servo motor 8-1, and the driving sprocket 8-2 is connected to the driven sprocket 8-4 through a chain 8-3. The driven sprocket 8-4 is rotatably sleeved on the fixed crossbar 7-1, and the driven sprocket 8-4 is fixedly connected to the square frame 7-3. The control line and power supply of the servo motor 8-1 extend backward to the rear side wall of the box body 1 and extend upward to be integrated into the microcomputer 17, and the speed of the servo motor 8-1 is controlled by the microcomputer 17. The rotation of the servo motor 8-1 drives the driven sprocket 8-4 to rotate, and then drives the square frame 7-3 to rotate, so that the radial magnetic field produces periodic changes.

[0035] The heating device 9 includes a heating tube 9-1. In this embodiment, the heating tube 9-1 is an electric heating tube. The heating tube 9-1 is horizontally arranged on the bottom plate of the box body 1 in an "S" shape. Branch pipes 9-2 are arranged at both ends of the heating tube 9-1. The branch pipes 9-2 are vertically arranged in the side plates of the box body 1. The tops of the branch pipes 9-2 extend to the top of the box body 1 and meet together. The side walls of the box body 1 where the branch pipes 9-2 are located are all provided with heat dissipation holes 9-3, embedded blowers 9-4 and air heaters 9-5. The embedded blowers 9-4 and the air heaters 9-5 send hot air to the center of the culture chamber 4 through the heat dissipation holes 9-3 on the two side walls. The coordinated use of the heating tube 9-1, the branch pipes 9-2, the heat dissipation holes 9-3, the embedded blowers 9-4 and the air heaters 9-5 can better keep the temperature in the box uniform.

[0036] The working principle of the present invention is as follows:

[0037] First, the cell culture plate is embedded in the base 7-2, and the two grooves 7-4 of the frame 7-3 are embedded with magnets to provide a radial magnetic field, and the annular magnet 7-5 provides an axial magnetic field. The servo motor 8-1 is controlled by the microcomputer 17 to rotate, and the rotation of the servo motor 8-1 drives the driven sprocket 8-4 to rotate, thereby driving the frame 7-3 to rotate, so that the magnetic field intensity generated by the magnet in the groove 7-4 remains unchanged, and the direction changes periodically, thereby allowing the nano magnetic particles to move under the traction of the magnetic field.

[0038] The heat emitted by the heating tube 9-1 in the heating chamber 6 is transferred upward through the branch pipe 9-2, and the hot air is sent to the center of the culture chamber 4 through the heat dissipation hole 9-3 and the embedded blower 9-4 to provide the required temperature for cell culture. The temperature and humidity in the box body 1 can be monitored in real time with the help of the temperature sensor 13 and the humidity sensor 14. With the help of the air inlet 10 and the air outlet 11, different gases can be connected externally according to the culture requirements, and the input speed of the external gas can be adjusted by detecting the air components in the incubator with the help of the gas sensor 15.

[0039] Embodiment 2

[0040] This example explores the ability of nanoparticles to penetrate cell spheroids under an external magnetic field, as described below:

[0041] Panc-1 and HPSC mixed cell spheres with fluorescence labels were cultured in vitro (green fluorescence is Calcein AM, marking HPSC cells; blue fluorescence is DAPI, marking Panc-1 cells), and then the obtained nanoparticles with CY3 red fluorescence labeling were dissolved in DMSO and added to the microporous culture dish for culturing cell spheres at a concentration of 0.5 mg / mL. The control group was cultured under incubator conditions without the influence of external field strength. The experimental group used a homemade external alternating field strength and incubated the cell spheres under the same culture conditions (37°C, 5% CO2) for 48 hours, and then observed with a two-photon microscope. The data processing results showed that the nanoparticles could not penetrate the cell spheres and enter their interior without external field strength (such as Figure 5 As shown in the figure), the internal structure of the cell sphere can be entered under the laboratory-made external field strength device (as shown in the figure). Figure 6 The fluorescence intensity of the fluorescence photos was analyzed using ImageJ software. It was found that when the external magnetic field device was not used, the red fluorescence intensity was low in the blue-green common area (cell sphere area), indicating that the nanoparticles failed to penetrate the cell sphere; after the external magnetic field device was used, the red fluorescence intensity filled the entire area, indicating that the nanoparticles successfully penetrated the matrix barrier and entered the cell sphere, verifying the penetration effect of the nanoparticles under the external magnetic field.

[0042] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. An incubator for driving magnetic nanoparticles to move in tissue cells, comprising a box body (1) and a box door (2), wherein the box body (1) is rotatably connected to the box door (2), and characterized in that: Two partitions (3) are arranged at the lower part of the box (1); a culture chamber (4) is formed between the upper partition (3) and the top plate of the box (1); a driving chamber (5) is formed between the two partitions (3); and a heating chamber (6) is formed between the lower partition (3) and the bottom plate of the box (1); The culture chamber (4) is provided with a magnetic driving device (7), the driving chamber (5) is provided with a power mechanism (8), and the magnetic driving device (7) is power-connected to the power mechanism (8); The heating chamber (6) is provided with a heating device (9), and the heating device (9) is connected to the culture chamber (4) through a pipeline.

2. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 1, characterized in that: The magnetic driving device (7) comprises a fixed cross bar (7-1), both ends of which are fixedly connected to the inner wall of the box body (1), a base (7-2) is fixedly connected to the fixed cross bar (7-1), a square frame (7-3) is arranged on the periphery of the base (7-2), the square frame (7-3) is rotatably connected to the fixed cross bar (7-1), grooves (7-4) are arranged on the inner sides of two frame frames of the square frame (7-3) parallel to the fixed cross bar (7-1), magnets are arranged in the grooves (7-4), and the two grooves (7-4) face one side of the base (7-2); The square frame (7-3) is in power connection with the power mechanism (8); A detachable annular magnet (7-5) is sleeved on the fixed crossbar (7-1), and the annular magnet (7-5) is located between the base (7-2) and the square frame (7-3).

3. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 2, characterized in that: The square frame (7-3) is rotatably connected to the fixed crossbar (7-1) via a bearing (7-6).

4. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 2, characterized in that: The power mechanism (8) comprises a servo motor (8-1), the servo motor (8-1) is arranged on the partition (3) located at the lower side, a driving sprocket (8-2) is fixedly connected to the output shaft of the servo motor (8-1), and the driving sprocket (8-2) is connected to the driven sprocket (8-4) through a chain (8-3); The driven sprocket (8-4) is rotatably sleeved on the fixed crossbar (7-1), and the driven sprocket (8-4) is fixedly connected to the square frame (7-3).

5. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 1, characterized in that: The heating device (9) comprises a heating pipe (9-1), the heating pipe (9-1) being arranged horizontally on the bottom plate of the box (1), branch pipes (9-2) being arranged at both ends of the heating pipe (9-1), the branch pipes (9-2) being arranged vertically in the side plates of the box (1), and the top ends of the branch pipes (9-2) extending to and meeting the top of the box (1); The side walls of the box body (1) where the branch pipe (9-2) is located are provided with heat dissipation holes (9-3), a built-in blower (9-4) and an air heater (9-5).

6. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 5, characterized in that: The heating tube (9-1) is arranged in an "S" shape.

7. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 1, characterized in that: An air inlet (10) and an air outlet (11) are provided on one side plate of the box body (1); A water vapor delivery port (12) is provided on the back plate of the box body (1); A temperature sensor (13), a humidity sensor (14), and a gas sensor (15) are arranged on the inner side of the top plate of the box body (1); A lamp tube (16) is detachably connected to the inner side of the top plate of the box body (1); The box door (2) is provided with an observation window (19).

8. The incubator for driving magnetic nanoparticles to move in tissue cells according to claim 7, characterized in that: A microcomputer (17) and a multifunctional display screen (18) are arranged on the upper side of the box (1); The temperature sensor (13), the humidity sensor (14), the gas sensor (15) and the power mechanism (8) are controlled by the microcomputer (17).

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