Stereoscopic culture separator for mesenchymal stem cells

Abstract

The invention relates to the technical field of cell culture and separation, in particular to a mesenchymal stem cell three-dimensional culture separator which comprises a base, an outer cylinder, a mounting frame and a motor and further comprises a transmission assembly, a connecting assembly, a culture plate and a beating assembly. The problems that when cells are separated from the culture plate, stress at all positions of the culture plate is imbalanced, the vibration effects of different positions on the culture plate are different, the cell separation effects are different, the vibration force of partial areas is too small, so that the cells are not thoroughly separated, or the vibration force is too large, so that the cells are broken are solved; the culture plate is movably connected with the connecting assembly, after one end of the culture plate is impacted and vibrates, the culture plate is turned over by 180 degrees through the transmission assembly and the connecting assembly, and then the hit point of the culture plate and the connecting position of the culture plate and the connecting assembly are switched; therefore, the similar vibration effect can be obtained at each position on the culture plate, and the cell separation effect is further ensured.

Description

Technical Field

[0001] The invention relates to the technical field of cell culture and separation, in particular to a three-dimensional culture and separation machine for mesenchymal stem cells. Background Art

[0002] Mesenchymal stem cells are a type of cell with multiple differentiation potentials. With the continuous development of cell culture technology, a variety of three-dimensional culture methods have been established. Compared with the single-layer adherent growth culture method of cells in two-dimensional culture, three-dimensional culture can fully simulate the in vivo environment of the organism, and the cells can grow in a three-dimensional spatial structure through a three-dimensional carrier. This culture method that simulates the in vivo environment has become the main method of cell culture at present.

[0003] After the culture of mesenchymal stem cells is completed, the cells need to be separated from the three-dimensional carrier scaffold. Centrifugation is usually used to separate the cells from the scaffold using strong centrifugal force. However, when the centrifugal speed is too fast, it is easy to damage the cells and even cause cell rupture. When the centrifugal speed is too slow, the ideal separation effect cannot be achieved.

[0004] In view of the above problems, some solutions have been proposed in the prior art, such as a rotary mesenchymal stem cell three-dimensional culture separator with publication number CN116731861A. This prior art generates a vibration effect through collision, thereby separating the cells adsorbed on the elastic culture plate; however, this prior art still cannot achieve the ideal separation effect. When the elastic plates collide, the vibration amplitude of the end of the elastic plate away from the connection is greater than the vibration amplitude of the connection, which makes the cell separation effect at one end of the elastic plate not ideal. If the rotation of the elastic culture plate is accelerated to give the connection of the elastic culture plate sufficient vibration force, the vibration frequency of the end of the elastic culture plate away from the connection will be too high, thereby causing damage to the cells.

[0005] To this end, a three-dimensional culture and separation machine for mesenchymal stem cells is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide a three-dimensional culture separation machine for mesenchymal stem cells, which solves the problem that when cells are separated from a culture plate by vibration force, different forces at different positions of the culture plate lead to different cell separation effects. By cyclically switching the fixed point and the struck position of the culture plate, a more uniform vibration force is provided for different positions of the culture plate, thereby ensuring the cell separation effect at each position of the culture plate.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] A mesenchymal stem cell three-dimensional culture and separation machine, comprising a base, an outer cylinder, a mounting frame and a motor, wherein the outer cylinder is connected to the base, the mounting frame is connected to the upper side of the base, the motor is arranged at the bottom of the base and connected to the outer cylinder, and further comprising a transmission assembly, a connecting assembly, a culture plate and a striking assembly, wherein the transmission assembly is connected to the base and the outer cylinder, the connecting assembly is arranged on the mounting frame and connected to the transmission assembly, the culture plate is connected to the connecting assembly, and the striking assembly is connected to the inner wall of the outer cylinder;

[0009] When the motor drives the outer cylinder to rotate from left to back and then to right, the striking assembly stirs the water flow and collects floating objects;

[0010] When the motor drives the outer cylinder to rotate from left to front and then to right, the striking assembly hits the bottom of the culture plate. The outer cylinder rotates through the transmission assembly to drive the connecting assembly to flip 180 degrees, and the culture plate moves down and changes the connection position with the connecting assembly.

[0011] Stem cells have the characteristic of adhering to the wall for growth. The growth environment in the body is simulated by a three-dimensional culture scaffold. After the cell culture is completed, the cells need to be separated from the culture scaffold. Usually, the culture plate can be vibrated to separate the cells. However, in the traditional scheme, the struck point of the culture plate and the connection point of the installation are usually not in the same position, and the vibration forces received by the two are also different, resulting in different separation effects. The separation effect of the connection point of the culture plate is often not as good as the separation effect of the struck point, which can easily lead to incomplete cell separation at a certain point of the culture plate, or excessive force at a certain point causing cell rupture. Through the above scheme, during the cell separation stage, the striking component follows the outer cylinder to rotate in the specified direction, thereby continuously striking the culture plate. The bottom of the culture plate, at this time, the bottom of the culture plate is the struck point, and the upper part connected to the connecting component is the connecting point. At this time, the vibration intensity generated by the struck point of the culture plate is higher than that of the upper connecting point, which results in that the cell separation effect on the lower side of the culture plate is better than that on the upper side; in order to balance the force and vibration effect of the culture plate, during the continuous rotation of the outer cylinder, the culture plate is regularly turned over at a specified frequency through the cooperation of the transmission component and the connecting component, and the turned culture plate will slide down under the influence of gravity. At this time, the part connected to the connecting component is the struck point in the previous step, thereby completing the cyclic exchange of the struck point and the connecting point, thereby achieving balanced force on the culture plate during the cell separation process, and ensuring the separation effect of cells at different positions on the culture plate;

[0012] During the cell culture stage, the culture plate is in a stationary state. At this time, the striking component slowly rotates with the outer cylinder and does not contact the culture plate. During the movement of the striking component, floating objects in the culture solution will be collected and sealed, thereby preventing too many floating objects from accumulating near the cells and affecting the normal growth of the cells.

[0013] Preferably, the transmission assembly includes a ratchet, a transmission gear, a driven gear, a fixed rod and a bevel gear 1, the ratchet is connected to the outer cylinder, the ratchet is a one-way structure, and the upper side is the observation angle. When the outer cylinder rotates clockwise, the ratchet will not cooperate with the transmission assembly. On the contrary, when the outer cylinder rotates counterclockwise, the ratchet will drive the transmission gear to move; the flipping action of the culture plate needs to be performed after the striking assembly hits, so the transmission of the ratchet and the transmission gear cannot be continuous, the coverage angle of the ratchet is less than one hundred and eighty degrees, and the culture plate can be flipped and the corresponding time is reserved for the striking assembly to hit the culture plate. The transmission gear is connected to the base and meshed with the ratchet, the driven gear is connected to the base and meshed with the transmission gear, the fixed rod is arranged in the mounting frame and connected to the driven gear, the bevel gear 1 is connected to the upper end of the fixed rod, and the bevel gear 1 is connected to the connecting assembly. When the outer cylinder rotates counterclockwise, the power will be transmitted to the connecting assembly through the above structure, thereby controlling the flipping of the connecting assembly.

[0014] Preferably, the connecting assembly includes bevel gear 2, a connecting rod, a connecting frame and a connecting groove, the bevel gear 2 is meshed with bevel gear 1, the connecting rod is connected to bevel gear 2, the connecting rod is rotatably connected to one side of the mounting frame, the connecting frame is connected to the connecting rod, the connecting groove is arranged on the connecting frame, the culture plate is connected in the connecting groove, when the outer cylinder drives bevel gear 1 to rotate a specified angle, bevel gear 2 connected to bevel gear 1 will drive the connecting frame to flip one hundred and eighty degrees, thereby turning the culture plate upside down, and then the culture plate slides down along the connecting groove under the action of gravity, so as to achieve the interchange of the struck point and the connecting point, the width of the connecting groove is greater than the width of the culture plate, and an elastic buffer pad is connected in the connecting groove, so that the culture plate has sufficient moving space in the connecting groove, on the one hand, reducing the resistance of the culture plate when it slides down, and on the other hand, when the culture plate is hit, the culture plate will tilt and collide with the connecting groove, thereby applying a certain vibration force at the connection point, thereby enhancing the cell separation effect.

[0015] Preferably, telescopic protrusions are provided at both upper and lower ends of the connecting rod, and a limiting groove is provided at the connection between the mounting frame and the connecting rod, and both sides of the limiting groove are inclined surfaces.

[0016] In order to ensure that there is no deviation when the culture plate is flipped, some limiting structures can be used to ensure its position. Through the above scheme, when the connecting rod is stationary, the telescopic protrusion connected to it will protrude from the surface of the connecting rod and sink into the limiting groove, thereby preventing its position from being offset by enhancing the resistance required for its offset.

[0017] Preferably, fixing strips are provided at both upper and lower ends of the culture plate, and the fixing strips are detachably connected to the culture plate. The fixing strips and the culture plate can be connected by bolts or clamping. When the culture plate is installed in the connecting groove, the fixing strip is first installed at one end of the culture plate, and then the end without the fixing strip is inserted into the connecting groove, and finally the fixing strip is installed at the end without the fixing strip. The fixing strip is semicircular, and the diameter of the fixing strip is larger than the width of the connecting groove. On the one hand, it is convenient to take it during installation and disassembly, and on the other hand, the contact between the fixing strip and the connecting groove can prevent the culture plate from falling. The fixing strip is made of elastic material, and after being hit by the striking component, the fixing strip itself will shake, thereby enhancing the vibration effect of the culture plate.

[0018] Preferably, the striking assembly comprises a fixed frame, a movable frame, a hinge seat, a hinge plate and a baffle, the fixed frame is connected to the outer cylinder, if the length direction of the fixed frame and the radial line of the outer cylinder are on the same straight line, then the contact surface when colliding with the culture plate will be relatively small, the fixed frame is inclined relative to the tangent of the inner wall of the outer cylinder, such a setting can make the culture plate receive a more balanced impact force in its length direction, the movable frame is clamped on the fixed frame, the hinge seat is arranged on the upper side of the movable frame, and the hinge plate is connected to the hinge seat, because different positions of the movable frame overlap with the culture plate at different times in the vertical direction, the side farther away from the outer cylinder contacts the culture plate faster, and correspondingly, the side closer to the outer cylinder contacts the culture plate slower;

[0019] There are multiple hinged plates along the length direction of the movable frame. Since there are multiple hinged plates, in addition to being able to achieve collision with the culture plate to a greater extent, multiple collisions with the culture plate can be achieved due to the different collision times between each hinged plate and the culture plate, thereby enhancing the vibration effect of the culture plate and thereby enhancing the cell separation effect. The baffle is connected to the movable frame and is arranged on a side of the hinged plate close to the fixed frame, with the upper side as the observation angle. When the outer cylinder drives the fixed frame to rotate clockwise, the hinged plate deflects in the opposite direction of the movement under the action of water flow resistance. At this time, the height of the hinged plate in the space is not enough to collide with the culture plate. When the outer cylinder drives the fixed frame to rotate counterclockwise, the hinged plate will also deflect in the opposite direction of the movement, but it will be blocked by the baffle during the deflection, so that it cannot deflect. At this time, the height of the hinged plate in the space is enough to collide with the culture plate.

[0020] Preferably, a striker is connected to the upper end of the hinge plate, the cross-section of the striker is circular, the diameter of the striker is greater than the width of the hinge plate, and the striker is made of elastic material.

[0021] The impact of the striking component on the culture plate is essentially a collision between the impact rod and the fixed bar. Through the above scheme, the impact rod is set to an elastic material. When it collides with the fixed bar, it will not cause damage to each other, and can provide sufficient impact force, thereby causing the culture plate to vibrate, thereby ensuring the cell separation effect; in addition, since the movable frame can be disassembled relative to the fixed frame, the diameter of the impact rod is larger than the width of the hinged plate. When disassembling, the impact rod can be lifted out by hand to facilitate the flushing of floating objects in the movable frame.

[0022] Preferably, the fixed frame is provided with a through slot, and the thickness of the through slot decreases continuously along the direction approaching the movable frame.

[0023] Through the above scheme, the thickness of the through groove decreases continuously in the direction close to the movable frame, and the curvature changes smoothly, which can reduce the resistance during movement and accelerate the flow rate of water entering the area. The expansion of the opening can facilitate the collection of floating objects.

[0024] Preferably, a filter hole is provided on one side of the movable frame away from the fixed frame, and a one-way plate is provided on the other side of the movable frame.

[0025] Through the above scheme, with the upper side as the observation angle, when the outer cylinder drives the fixed frame to rotate clockwise, the water flows into the movable frame through the through groove, and then flows out through the filter hole. At this time, the floating objects will be blocked by the filter hole and remain in the movable frame, and due to the effect of the one-way plate, when the water flow direction is opposite, the one-way plate is closed to prevent the floating objects from running out.

[0026] Preferably, guide grooves are provided at upper and lower positions of the fixed frame and the movable frame, the directions of the guide grooves are perpendicular to the length directions of the fixed frame and the movable frame, and the guide grooves are semicircular.

[0027] Through the above scheme, the setting of the guide groove can guide the flow of the culture solution when it moves, which can enhance the flow effect of the culture solution in the outer tube and reduce the resistance when the fixed frame and the movable frame move.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] 1. The present invention solves the problem that when separating cells from a culture plate, the forces at various positions on the culture plate are uneven, resulting in different vibration effects at different positions on the culture plate, different cell separation effects, and incomplete cell separation due to too small vibration force in some areas, or cell rupture due to too large vibration force. The culture plate is movably connected to a connecting component, and when one end of the culture plate is hit and vibrates, the culture plate is flipped 180 degrees through a transmission component and a connecting component, and then the struck point of the culture plate and the connection position with the connecting component are switched, thereby ensuring that similar vibration effects can be obtained at various positions on the culture plate, thereby ensuring the cell separation effect.

[0030] 2. By setting up a striking component, during the cell culture stage, the striking component rotates with the outer cylinder, continuously filters the culture solution during the rotation process, collects and seals the floating objects in the culture solution, thereby preventing too many floating objects from accumulating near the cells and affecting the normal growth of the cells; during the cell separation stage, the striking component can hit the bottom of the culture plate, causing the culture plate to vibrate, thereby achieving separation of the cells from the culture plate.

[0031] 3. By setting the width of the connecting groove to be larger than the width of the culture plate, on the one hand, when the culture plate is flipped upside down and slides down, the resistance during the movement of the culture plate can be reduced to prevent the connecting groove from scratching the surface of the culture plate; on the other hand, when the bottom of the culture plate is hit, the culture plate will tilt to a certain extent, and the contact position with the connecting groove will also collide, thereby generating vibration on the lower side and providing a certain degree of vibration force at the connection, thereby enhancing the cell separation effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 It is a cross-sectional view of the structure of the present invention as a whole;

[0033] Figure 2 It is a structural schematic diagram of the transmission assembly of the present invention;

[0034] Figure 3 It is a structural schematic diagram of the connection position between the connection assembly and the mounting frame of the present invention;

[0035] Figure 4 It is a structural schematic diagram of the connection assembly of the present invention;

[0036] Figure 5 It is a structural schematic diagram of the culture plate of the present invention;

[0037] Figure 6 It is a structural schematic diagram of the striking assembly of the present invention;

[0038] Figure 7 It is a structural schematic diagram of the striking assembly of the present invention from another side perspective;

[0039] Figure 8 This is a diagram showing the tilted state of the hinged plate during the cell culture stage of the present invention.

[0040] In the figure: 1. base; 2. outer cylinder; 3. mounting frame; 4. motor; 5. transmission assembly; 501. ratchet; 502. transmission gear; 503. driven gear; 504. fixing rod; 505. bevel gear one; 6. connecting assembly; 601. bevel gear two; 602. connecting rod; 603. connecting frame; 604. connecting groove; 605. elastic buffer pad; 606. telescopic protrusion; 607. limiting groove; 7. culture plate; 8. striking assembly; 801. fixing frame; 802. movable frame; 803. hinged seat; 804. hinged plate; 805. baffle; 806. impact rod; 807. through groove; 808. filter hole; 809. one-way plate; 8010. guide groove; 9. fixing strip. DETAILED DESCRIPTION

[0041] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0042] See also Figures 1 to 8 The present invention provides a three-dimensional culture and separation machine for mesenchymal stem cells, and the technical solution is as follows:

[0043] For details, please refer to Figures 1 to 4 A mesenchymal stem cell three-dimensional culture and separation machine comprises a base 1, an outer cylinder 2, a mounting frame 3 and a motor 4. The outer cylinder 2 is connected to the base 1, and a leg is fixed under the base 1. The mounting frame 3 is connected to the upper side of the base 1. The mounting frame 3 is a rectangular parallelepiped structure, and its interior is a hollow structure that can accommodate part of the structure of a transmission component 5. The motor 4 is arranged at the bottom of the base 1 and is connected to the outer cylinder 2. The base 1 is only used to fix the installation of the motor 4 and is not driven by the motor 4. The motor 4 can only drive the outer cylinder 2 to rotate clockwise or counterclockwise. It also comprises a transmission component 5, a connecting component 6, a culture plate 7 and a striking component 8. The transmission component 5 is connected to the base 1 and the outer cylinder 2. The transmission component 5 It includes a ratchet 501, a transmission gear 502, a driven gear 503, a fixing rod 504 and a bevel gear 505. The ratchet 501 is a one-way tooth structure. The ratchet 501 is connected to the inner wall of the outer cylinder 2. Two transmission gears 502 are respectively arranged on the left and right sides of the mounting frame 3. The transmission gear 502 is connected to the base 1 through a bearing, and the transmission gear 502 can mesh with the ratchet 501. When viewed from a top view, in the cell culture stage, the outer cylinder 2 rotates clockwise. At this time, the ratchet 501 cannot drive the transmission component 5 to rotate due to its one-way tooth structure. In the cell separation stage, the outer cylinder 2 rotates counterclockwise. At this time, the ratchet 501 will drive the transmission gear 502 to rotate.

[0044] After the culture plate 7 is turned over, a certain amount of time needs to be reserved for the striking assembly 8 to strike the culture plate 7, so the coverage angle of the ratchet 501 is less than 180 degrees, which is equivalent to that the ratchet 501 cannot mesh with the transmission gears 502 on the left and right sides at the same time. This arrangement makes it possible for the ratchet 501 to mesh with the next transmission gear 502 on the path only after the striking assembly 8 strikes the culture plate 7, thereby controlling the culture plate 7 to turn over;

[0045] The driven gear 503 is connected to the base 1 through a bearing, the driven gear 503 and the transmission gear 502 are meshed with each other, the fixing rod 504 is fixed at the center position of the upper side of the driven gear 503, and the upper side of the fixing rod 504 is located inside the fixing frame 801, the bevel gear 1 505 is connected to the upper end of the fixing rod 504, and the bevel gear 1 505 is connected to the connecting component 6, and the two connecting components 6 are respectively arranged on the left and right sides of the mounting frame 3. Described below is a connecting assembly 6 arranged on the left side of the mounting frame 3, the connecting assembly 6 includes a bevel gear 2 601, a connecting rod 602, a connecting frame 603 and a connecting groove 604, the bevel gear 2 601 is meshed with the bevel gear 1 505, the connecting rod 602 is fixed to the left side of the bevel gear 2 601, the connecting rod 602 is rotatably connected to the mounting frame 3, a slot for accommodating the connecting rod 602 is provided on the mounting frame 3, two telescopic protrusions 606 are provided at the connection between the connecting rod 602 and the mounting frame 3, the two telescopic protrusions 606 are respectively installed at the upper and lower ends of the connecting rod 602, the telescopic protrusion 606 will be retracted into the inside of the connecting rod 602 when squeezed, otherwise it will pop out of the surface of the connecting rod 602, correspondingly, limiting grooves 607 are provided on the upper and lower sides of the connection between the mounting frame 3 and the connecting rod 602, the limiting grooves 607 can accommodate the telescopic protrusions 606 protruding from the surface of the connecting rod 602, and inclined surfaces are provided on both sides of the limiting grooves 607, which can facilitate the rotation of the culture plate 7 when it is necessary to flip it;

[0046] The connecting frame 603 is fixed to the left end of the connecting rod 602. The connecting frame 603 is a rectangular parallelepiped as a whole. A connecting groove 604 is provided in the middle of the connecting frame 603. The connecting groove 604 is also a cube. The culture plate 7 is connected in the connecting groove 604, and the culture plate 7 can move up and down in the connecting groove 604. The width of the connecting groove 604 is greater than the width of the culture plate 7. After the bottom of the culture plate 7 is hit, the position of the culture plate 7 close to the connecting groove 604 will also tilt and collide with the inner wall of the connecting groove 604, and then a certain vibration will also be generated at a position far away from the hitting point, thereby ensuring the cell separation effect. An elastic buffer pad 605 is connected to the connecting groove 604. The elastic buffer pad 605 is made of rubber. The elastic buffer pad 605 can cover the four corners of the connecting groove 604 to prevent the culture plate 7 from being scratched.

[0047] As an embodiment of the present invention, refer to Figure 5, the culture plate 7 is provided with many slots for accommodating three-dimensional culture brackets. The three-dimensional culture bracket can adopt a three-dimensional structural carrier composed of a nanofiber bracket, a porous bracket three-dimensional culture or a three-dimensional hydrogel bracket, etc., and the upper and lower ends of the culture plate 7 are provided with fixing bars 9, and the fixing bar 9 and the culture plate 7 are detachably connected. The fixing bar 9 and the culture plate 7 can be connected by a snap-on method, so that they can be easily installed and replaced. If the stability of the connection is sought, the fixing bar 9 and the culture plate 7 can be fixed by bolts. The fixing bar 9 is semicircular, and the side with the arc is installed close to the culture plate 7. The fixing bar 9 is made of elastic material. When it is hit by the striking component 8, it will vibrate, thereby ensuring the vibration separation effect of the culture plate 7. The diameter of the fixing bar 9 is larger than the width of the connecting groove 604, so it can be conveniently taken when it is installed and disassembled.

[0048] As an embodiment of the present invention, refer to Figures 6 to 8 The striking assembly 8 is connected to the inner wall of the outer tube 2. The striking assembly 8 includes a fixed frame 801, a movable frame 802, a hinge seat 803, a hinge plate 804 and a baffle 805. The fixed frame 801 is connected to the outer tube 2. The fixed frame 801 is inclined relative to the tangent of the inner wall of the outer tube 2, which can increase the contact area between the striking part and the culture plate 7 as much as possible, and also make the collision force on the culture plate 7 perpendicular to it, so as to avoid the dispersion of the collision force. The movable frame 802 is connected to the fixed frame 801 by clamping. The clamping of the two can be achieved by setting a clamping block on one side of the movable frame 802 and setting a clamping groove at the corresponding position of the fixed frame 801. The structure is simple and convenient for disassembly and assembly;

[0049] The hinge seat 803 is arranged on the upper side of the movable frame 802, and the hinge plate 804 is connected to the hinge seat 803. The hinge plates 804 are arranged in a plurality along the length direction of the movable frame 802. The side farther away from the outer cylinder 2 will contact the culture plate 7 faster, and correspondingly, the side closer to the outer cylinder 2 will contact the culture plate 7 slower, so that the plurality of hinge plates 804 can impact the culture plate 7 multiple times, thereby enhancing the vibration effect on the culture plate 7, thereby ensuring the cell separation effect. The baffle 805 is connected to the movable frame 802 and is arranged on the hinge plate 80 4 is close to the side of the fixing frame 801, which can prevent the hinged plate 804 from deviating toward the fixing frame 801. The upper end of the hinged plate 804 is connected with a striker 806. The cross section of the striker 806 is circular. The diameter of the striker 806 is larger than the width of the hinged plate 804. The movable frame 802 can be taken by taking the striker 806, which is convenient for installation and disassembly. The striker 806 is made of elastic material. When it collides with the fixing bar 9, it will not cause damage to each other, but can also provide sufficient impact force, thereby causing the culture plate 7 to vibrate, thereby ensuring the cell separation effect;

[0050] The fixed frame 801 is provided with a through groove 807, and the thickness of the through groove 807 decreases along the direction close to the movable frame 802, thereby reducing the resistance during movement and accelerating the flow rate of water entering the area. The expansion of the opening can facilitate the collection of floating objects. The movable frame 802 is provided with a filter hole 808 on the side away from the fixed frame 801, which can collect floating objects in the culture solution during the cell culture stage to prevent excessive floating objects from accumulating near the cells and affecting the normal growth of the cells. The other side of the movable frame 802 is provided with a single The one-way plate 809 is closed when the movable frame 802 moves in the reverse direction to prevent floating flow from flowing out; the upper and lower positions of the fixed frame 801 and the movable frame 802 are both provided with guide grooves 8010, and the direction of the guide grooves 8010 is perpendicular to the length direction of the fixed frame 801 and the movable frame 802, which can guide the flow of the culture solution when they move, can enhance the flow effect of the culture solution in the outer tube 2, and can also reduce the resistance when the fixed frame 801 and the movable frame 802 move, and the guide grooves 8010 are semicircular.

[0051] The specific working principle is: during the cell culture stage, the culture plate 7 remains stationary in order to properly stir the culture solution so that the nutrients in the culture solution can be evenly distributed in the culture solution, and then the cells can fully absorb the nutrients in the culture solution. The upper side is used as the observation angle. At this time, the outer cylinder 2 rotates clockwise under the drive of the motor 4, and the fixed frame 801 connected to the outer cylinder 2 will act as a paddle. At this time, the fixed frame 801 follows the outer cylinder 2 to move at a constant speed to stir the culture solution.

[0052] At the same time, the fixed frame 801 will move with the movable frame 802. Due to the effect of water flow resistance, the hinged plate 804 will tilt in the direction away from the fixed frame 801, so the height of the hinged plate 804 in the space is reduced, and no collision will occur when the hinged plate 804 passes the bottom of the culture plate 7.

[0053] Since cells will inevitably produce some floating objects during the culture process, these floating objects will float in the culture solution. If the floating objects near the cells accumulate too much, it is easy to affect the normal absorption of nutrients in the culture solution by the cells, thereby affecting the quantity and quality of cell culture. In order to eliminate this effect, it is necessary to reduce the number of floating objects and remove them in time. When the fixed frame 801 and the movable frame 802 rotate clockwise, the water flow of the culture solution will pass through the through groove 807 and the one-way plate 809 into the movable frame 802, and the floating objects in the water flow will be blocked by the filter hole 808, thereby reducing the floating objects near the cells. The design of the through groove 807 gradually reducing its thickness along the direction close to the movable frame 802 can speed up the flow rate of the culture solution into the through groove 807, thereby enhancing the collection effect of floating objects.

[0054] In the cell separation stage, the striker 806 needs to hit the fixed bar 9 to vibrate the culture plate 7, thereby separating the cells from the culture support in the culture plate 7. With the upper side as the observation angle, the outer cylinder 2 rotates counterclockwise under the drive of the motor 4, and the hinged plate 804 moves toward the fixed frame 801 under the action of water flow resistance. However, due to the existence of the baffle 805, the hinged plate 804 is in a vertical state. At this time, the striker 806 on the hinged plate 804 is high enough in the space to hit the fixed bar 9 on the lower side of the culture plate 7. Therefore, after the collision between the two, the culture plate 7 will vibrate, thereby separating the cells from the culture plate 7.

[0055] Due to the limitation of the striking point and the connection position of the culture plate 7, the vibration effect near the striking point is better than the effect at the connection position of the culture plate 7, and the cell separation effects of the two will also be different. In order to eliminate this effect, the striking point and the connection point of the culture plate 7 can be replaced cyclically. After the culture plate 7 is hit once, the ratchet 501 connected to the outer cylinder 2 will mesh with the transmission gear 502, and then the connecting frame 603 will be turned over 180 degrees through the driven gear 503, the fixing rod 504, the bevel gear 1 505, the bevel gear 2 601 and the connecting rod 602. After the connecting frame 603 is turned over 180 degrees, the culture plate 7 will slide down along the connecting groove 604 under the influence of gravity, thereby realizing the switching of the connection point and the striking point. After the switching, the next striking can be performed to ensure the uniformity of vibration at each position of the culture plate 7, thereby ensuring the cell separation effect.

[0056] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A mesenchymal stem cell three-dimensional culture and separation machine, comprising a base (1), an outer cylinder (2), a mounting frame (3) and a motor (4), wherein the outer cylinder (2) is connected to the base (1), the mounting frame (3) is connected to the upper side of the base (1), and the motor (4) is arranged at the bottom of the base (1) and connected to the outer cylinder (2), characterized in that: It also comprises a transmission assembly (5), a connection assembly (6), a culture plate (7) and a striking assembly (8), wherein the transmission assembly (5) is connected to the base (1) and the outer cylinder (2), the connection assembly (6) is arranged on the mounting frame (3) and connected to the transmission assembly (5), the culture plate (7) is connected to the connection assembly (6), and the striking assembly (8) is connected to the inner wall of the outer cylinder (2); When the motor (4) drives the outer cylinder (2) to rotate from left to back and then to right, the striking assembly (8) stirs the water flow and collects floating objects; When the motor (4) drives the outer cylinder (2) to rotate from left to front and then to right, the striking assembly (8) strikes the bottom of the culture plate (7), and the outer cylinder (2) rotates through the transmission assembly (5) to drive the connecting assembly (6) to flip 180 degrees, and the culture plate (7) moves downward and changes the connection position with the connecting assembly (6).

2. A three-dimensional culture and separation machine for mesenchymal stem cells according to claim 1, characterized in that: The transmission assembly (5) comprises a ratchet (501), a transmission gear (502), a driven gear (503), a fixed rod (504) and a bevel gear 1 (505); the ratchet (501) is connected to the outer cylinder (2); the coverage angle of the ratchet (501) is less than one hundred and eighty degrees; the transmission gear (502) is connected to the base (1) and meshes with the ratchet (501); the driven gear (503) is connected to the base (1) and meshes with the transmission gear (502); the fixed rod (504) is arranged in the mounting frame (3) and is connected to the driven gear (503); the bevel gear 1 (505) is connected to the upper end of the fixed rod (504); and the bevel gear 1 (505) is connected to the connecting assembly (6).

3. A three-dimensional culture and separation machine for mesenchymal stem cells according to claim 2, characterized in that: The connecting assembly (6) comprises a second bevel gear (601), a connecting rod (602), a connecting frame (603) and a connecting groove (604); the second bevel gear (601) is meshed with the first bevel gear (505); the connecting rod (602) is connected to the second bevel gear (601); the connecting rod (602) is rotatably connected to one side of the mounting frame (3); the connecting frame (603) is connected to the connecting rod (602); the connecting groove (604) is arranged on the connecting frame (603); the culture plate (7) is connected to the connecting groove (604); the width of the connecting groove (604) is greater than the width of the culture plate (7); and an elastic buffer pad (605) is connected inside the connecting groove (604).

4. A three-dimensional culture and separation machine for mesenchymal stem cells according to claim 3, characterized in that: The upper and lower ends of the connecting rod (602) are both provided with telescopic protrusions (606), and the connection between the mounting frame (3) and the connecting rod (602) is provided with a limiting groove (607), and both sides of the limiting groove (607) are inclined surfaces.

5. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 3, characterized in that: The culture plate (7) is provided with fixing strips (9) at both upper and lower ends. The fixing strips (9) are detachably connected to the culture plate (7). The fixing strips (9) are semicircular and made of elastic material. The diameter of the fixing strips (9) is greater than the width of the connecting groove (604).

6. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 1, characterized in that: The striking assembly (8) comprises a fixed frame (801), a movable frame (802), a hinge seat (803), a hinge plate (804) and a baffle (805); the fixed frame (801) is connected to the outer tube (2); the fixed frame (801) is arranged to be inclined relative to the tangent of the inner wall of the outer tube (2); the movable frame (802) is clamped on the fixed frame (801); the hinge seat (803) is arranged on the upper side of the movable frame (802); the hinge plate (804) is connected to the hinge seat (803); a plurality of hinge plates (804) are arranged along the length direction of the movable frame (802); and the baffle (805) is connected to the movable frame (802) and is arranged on a side of the hinge plate (804) close to the fixed frame (801).

7. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 6, characterized in that: The upper end of the hinged plate (804) is connected to a striker rod (806), the cross section of the striker rod (806) is circular, the diameter of the striker rod (806) is greater than the width of the hinged plate (804), and the striker rod (806) is made of elastic material.

8. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 6, characterized in that: The fixed frame (801) is provided with a through slot (807), and the thickness of the through slot (807) decreases continuously along the direction approaching the movable frame (802).

9. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 6, characterized in that: A filter hole (808) is provided on one side of the movable frame (802) away from the fixed frame (801), and a one-way plate (809) is provided on the other side of the movable frame (802).

10. The three-dimensional culture and separation machine for mesenchymal stem cells according to claim 6, characterized in that: The fixed frame (801) and the movable frame (802) are both provided with guide grooves (8010) at upper and lower positions, the direction of the guide grooves (8010) is perpendicular to the length direction of the fixed frame (801) and the movable frame (802), and the guide grooves (8010) are semicircular.

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