Cell microsphere preparation device

Abstract

The application discloses a cell microsphere preparation device, which comprises a bolus pump assembly, a mixing assembly arranged adjacent to the bolus pump assembly and a spraying assembly; the bolus pump assembly is used for providing power to extrude mixed liquid in the mixing assembly from the spraying assembly; wherein the mixing assembly comprises a fixing frame, a roller rotatably connected to the fixing frame, a piston rod sliding in the roller and a driving gear engaged with the roller. According to the application, the cell concentration in the mixed suspension is ensured to be always uniform, thereby improving the preparation quality and batch stability of the cell microspheres and meeting the actual needs of the field of biopharmaceuticals and cell therapy for high-quality cell microsphere carriers.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, and in particular to an apparatus for preparing cell microspheres. Background Technology

[0002] In the fields of biopharmaceuticals, cell therapy, and tissue engineering, cell microspheres serve as core cell carriers, leveraging their protective effects on cells and controllable physicochemical properties to drive the industrialization of related technologies. Among them, calcium alginate cell microspheres, due to their excellent biocompatibility, biodegradability, and low immunogenicity, have become one of the most widely used types of cell microspheres.

[0003] Current methods for preparing cell microspheres mainly include emulsification, microfluidics, and electrostatic spraying (hereinafter referred to as "electroplated method"). Among them, the electrostatic spraying method has become the mainstream technology for large-scale preparation of calcium alginate cell microspheres due to its relatively simple equipment structure, convenient operation, high preparation efficiency, and strong controllability of microsphere size. Taking the preparation of calcium alginate cell microspheres by electrostatic spraying as an example, its typical preparation process is as follows: First, live cells are mixed with a sodium alginate aqueous solution of a specific concentration according to a preset ratio to prepare a homogeneous cell-sodium alginate mixed suspension; Second, the mixed suspension is injected into a storage container and connected to a spraying assembly with a metal needle through a conduit; Third, a high-voltage electrostatic field is applied to the spraying assembly, while the storage container is controlled to deliver the mixed suspension to the spraying assembly. Under the action of electrostatic repulsion, the mixed suspension overcomes the surface tension and splits into tiny droplets from the tip of the needle and is sprayed at high speed; Fourth, the sprayed droplets fall into a cross-linking liquid containing calcium ions, and sodium alginate and calcium ions are instantaneously cross-linked and solidified to form calcium alginate cell microspheres with stable structure and suitable mechanical strength.

[0004] However, existing cell microsphere preparation devices based on electro-spraying have significant technical drawbacks: the cell-sodium alginate mixed suspension is prone to sedimentation during preparation, leading to inhomogeneity in the mixture. Since cell density is typically higher than that of sodium alginate aqueous solution, under gravity, as the mixed suspension is stored in the reservoir, flows through the conduit, and enters the inlet of the spraying assembly, cells gradually settle to the bottom of the container, the inner wall of the conduit, and the inlet area. This sedimentation is particularly pronounced during longer preparation periods. This inhomogeneity directly results in extremely poor consistency of the prepared cell microspheres: on the one hand, the cell concentration in the sedimentation zone is too high, causing large clusters of cells within the microspheres. This not only hinders nutrient exchange and metabolic waste removal but also easily leads to cell hypoxia, nutrient deficiency, and inactivation. On the other hand, the cell concentration in the upper layer or the un-sedimented areas within the conduit is extremely low or even cellless, forming empty or low-cell-content microspheres, making it difficult to achieve the intended cell carrier function. These problems severely restrict the quality stability of cell microspheres, increase the difficulty of quality control in industrial production, and pose potential risks to their clinical application. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a cell microsphere preparation device. Addressing the technical pain points of sedimentation of the mixed suspension and poor microsphere uniformity in existing cell microsphere preparation devices, this invention provides a preparation device that can efficiently and continuously mix the cell-sodium alginate mixed suspension throughout the entire preparation process. This ensures that the cell concentration in the mixed suspension remains uniform, thereby improving the preparation quality and batch stability of cell microspheres and meeting the practical needs of the biopharmaceutical and cell therapy fields for high-quality cell microsphere carriers. To achieve the above-mentioned objectives and other advantages of the present invention, a cell microsphere preparation device is provided, comprising: A push pump assembly, a mixing assembly and a jetting assembly disposed adjacent to the push pump assembly; The injection pump assembly is used to provide power to expel the mixture from the mixing assembly from the jet assembly; The mixing assembly includes a fixed frame, a roller rotatably connected to the fixed frame, a piston rod sliding in the roller, and a drive gear meshing with the roller. The roller is provided with at least one piston rod.

[0006] Preferably, the roller includes a cylinder body, one end of which is integrally connected to a conveying pipe, and a conveying chamber is provided inside the cylinder body, which is connected to the conveying pipe.

[0007] Preferably, a ring of meshing teeth is fixed to the surface of the cylinder at the end away from the conveying pipe, and the meshing teeth mesh with the drive gear.

[0008] Preferably, the cylindrical body has a conical structure.

[0009] Preferably, the conveying chamber is cylindrical, and the angle between the axis of the conveying chamber and the center line of the cone is between 5° and 30°.

[0010] Preferably, the injection assembly includes an L-shaped nozzle, which is connected in communication with the inside of the delivery pipe.

[0011] Preferably, the injection pump assembly includes a slide rail, a slider slidably disposed on the slide rail, and a disc fixedly connected to one end of the slider, wherein a piston rod is abutted and connected to the disc.

[0012] Preferably, the end of the piston rod near the disk is spherical, the spherical end of the piston rod rotates on the surface of the disk, and the piston rod is inserted into the conveying chamber from the end of the disk.

[0013] Compared with the prior art, the advantages and positive effects of the present invention are: ensuring that the cell concentration in the mixed suspension is always uniform, thereby improving the preparation quality and batch stability of cell microspheres, and meeting the actual needs of the biopharmaceutical and cell therapy fields for high-quality cell microsphere carriers. Attached Figure Description

[0014] Figure 1 This is a three-dimensional structural schematic diagram of the cell microsphere preparation device according to the present invention; Figure 2 This is a three-dimensional exploded view of the apparatus for preparing cell microspheres according to the present invention; Figure 3 This is a three-dimensional structural diagram of the roller in the cell microsphere preparation apparatus according to the present invention; Figure 4 This is a schematic diagram of a half-section of the roller in the cell microsphere preparation apparatus according to the present invention; Figure 5 A schematic diagram of the overall structure of the drum of the cell microsphere preparation apparatus according to the present invention, which has two piston rods; Figure 6 A schematic diagram of a half-section of the drum of the cell microsphere preparation apparatus according to the present invention, having two piston rods; Figure 7 This is a schematic diagram of the piston rod and disk of the drum in the cell microsphere preparation apparatus according to the present invention; Figure 8 This is a half-sectional schematic diagram of the L-shaped nozzle of the drum in the cell microsphere preparation apparatus according to the present invention; Figure 9 This is a schematic diagram of the structure of a drum with two piston rods in the drum of the cell microsphere preparation apparatus according to the present invention, showing a cell cluster inside. Detailed Implementation

[0015] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0016] Reference Figure 1 Cell microsphere preparation apparatus, comprising: A base 20 is provided, on which a push pump assembly 10, a mixing assembly 20 and a spray assembly 30 are placed. The push pump assembly 10 is used to provide power to squeeze the mixture in the mixing assembly 20 out of the spray assembly 30. The push pump provides a pushing driving force to the mixing assembly, gradually squeezing the mixture of sodium alginate and cells in the mixing assembly out of the L-shaped nozzle and spraying it into a container under electrostatic action. The container contains a calcium chloride solution, and the sodium alginate reacts with calcium ions to form calcium alginate microspheres.

[0017] like Figure 2 As shown, the mixing assembly 20 includes a fixed frame 23, a roller 22 rotatably connected to the fixed frame 23, a piston rod 21 sliding within the roller 22, and a drive gear 24 meshing with the roller 22. The roller 22 includes a cylindrical body 221, one end of which is integrally connected to a conveying pipe 222. A conveying chamber 223 is formed inside the cylindrical body 221 and communicates with the conveying pipe 222. A ring of meshing teeth is fixedly attached to the surface of the cylindrical body 221 at the end away from the conveying pipe 222, and these meshing teeth mesh with the drive gear 24. A through hole is formed at the upper end of the fixed frame 23, and the roller 22 is rotatably placed within this through hole. In use, a drive motor is placed in the bottom of the fixed frame 23. The output shaft of the drive motor is fixedly connected to the drive gear 24. When the drive motor rotates, it drives the drive gear 24 to rotate. The drive gear 24 meshes with the meshing teeth on the roller 22, thereby driving the roller 22 to rotate, thus realizing that the roller 22 is in a stable rolling state.

[0018] like Figure 3-4As shown, the cylindrical body 221 has a conical structure, and its diameter gradually decreases along the direction of the meshing teeth towards the delivery pipe 222. The delivery chamber 223 is cylindrical, and the angle between the axis of the delivery chamber 223 and the center line of the cone is between 5° and 30°. The conical structure of the cylindrical body 221 is problematic because cell clusters in the sodium alginate solution tend to settle to the bottom due to gravity. This can lead to poor uniformity of the cell clusters encapsulated in calcium alginate during spraying. For example, there might be more cell clusters at the bottom, resulting in more cell clusters within the calcium alginate microspheres formed after spraying into the calcium alginate solution, while fewer cell clusters are formed at the top, leading to fewer or no cell clusters after spraying, similar to how some eggs have double yolks while others are hollow. Placing the delivery chamber 223 horizontally and rotating it circumferentially can effectively reduce the precipitation of cell clusters in the sodium alginate solution. However, the cells at the front and rear ends of the delivery chamber 223 are not mixed evenly, resulting in unevenness between the front and rear ends. Therefore, in this application, the delivery chamber 223 is arranged at an angle to the axis of rotation. Specifically, a conical mixing mechanism is designed so that when the delivery chamber 223 is rotated above the axis, the cells precipitate towards the front end of the delivery chamber 223, and when the delivery chamber 223 is rotated below the axis, the cells precipitate towards the rear end of the delivery chamber 223, thereby making the cell suspension uniform overall.

[0019] like Figure 2 As shown, the spray assembly 30 includes an L-shaped nozzle 31, which is internally connected to the delivery pipe 222. A mounting plate is fixed to the side of the mounting bracket 23 near the L-shaped nozzle 31. A mounting through hole is formed on the mounting plate, into which the L-shaped nozzle 31 is inserted to maintain stability. A container 33 is placed directly below the L-shaped nozzle 31, and a conductive ring 32 is positioned directly above the container 33, fixed to the mounting bracket 23. The container 33 contains a calcium chloride solution. Figure 8 As shown, the L-shaped nozzle 31 includes a connecting section 311, a vertical section 312 fixedly connected to the connecting section 311, and an ejection section 313 fixedly connected to the vertical section 312. The connecting section 311 has a hollow internal structure, and one end of the delivery pipe 222 is inserted into the connecting section 311, allowing the delivery pipe 222 to rotate within it. Both the vertical section 312 and the vertical section 312 have through holes, and their interiors are interconnected. The connecting section 311 and the vertical section 312 are arranged perpendicularly to each other, allowing the cell suspension to be ejected from the fixed nozzle during the rotation of the delivery chamber 223. The nozzle needs to be kept fixed, and the delivery chamber 223 needs to be able to rotate with the nozzle. The rotational engagement of the L-shaped nozzle changes the horizontal outlet of the L-shaped nozzle 31 to a vertical position, spraying the solution into the container.

[0020] like Figure 1As shown, the injection pump assembly 10 includes a motor mounting base, on which two lead screw mounting bases are fixedly connected. Two lead screws are rotatably connected between the two lead screw mounting bases, and a slider 12 is slidably connected to each lead screw. A slide rail 11, a slider 12 slidably disposed on the slide rail 11, and a disc 13 fixedly connected to one end of the slider 12, with a piston rod 21 abutting against the disc 13. Figure 7 As shown, the piston rod 21 includes a piston rod 211 and a spherical end 212 fixed to one end of the piston rod 211. The end of the piston rod 211 away from the spherical end 212 is inserted into the roller 22, and the spherical end 212 abuts against the surface of the disk 13. When the drive gear 24 drives the roller 22 to rotate, the spherical end 212 rotates on the surface of the disk 13. During operation, the motor drives the lead screw to rotate, which in turn drives the slider 12 to move towards the mixing assembly 20, causing the disk 13 to drive the piston rod 21 to move towards the roller 22. This causes the piston rod 21 to perform piston movement in the conveying chamber 223, squeezing the mixture in the conveying chamber 223 into the conveying pipe 222, thereby allowing the mixture to enter the L-shaped nozzle 31 and be sprayed into the container 33 under electrostatic action. The container 33 contains a calcium chloride solution, and sodium alginate reacts with calcium ions to form calcium alginate microspheres.

[0021] One embodiment of a cell microsphere preparation apparatus includes: like Figure 5 As shown, a base 20 is provided, on which a push pump assembly 10, a mixing assembly 20 adjacent to the push pump assembly 10, and a spray assembly 30 are placed. The push pump assembly 10 provides power to expel the mixture in the mixing assembly 20 from the spray assembly 30. The push pump provides a pushing driving force to the mixing assembly, gradually expelling the mixture of sodium alginate and cells from the mixing assembly through an L-shaped nozzle and spraying it into a container containing a calcium chloride solution under electrostatic action. The sodium alginate reacts with calcium ions to form calcium alginate microspheres. The mixing assembly 20 includes a fixed frame 23, a roller 22 rotatably connected to the fixed frame 23, a first piston rod 25 and a second piston rod 26 sliding in the roller 22, and a drive gear 24 meshing with the roller 22. Figure 6As shown, the roller 22 includes a cylindrical body 221, one end of which is integrally connected to a conveying pipe 222. The cylindrical body 221 has a first conveying chamber 224 and a second conveying chamber 225 inside. The conveying chamber 223 and the end of the conveying pipe 222 near the pipe 222 are interconnected to form a conveying section, which is connected to the conveying pipe 222. A first piston rod 25 and a second piston rod 26 are slidably disposed in the first conveying chamber 224 and the second conveying chamber 225, respectively. A ring of meshing teeth is fixedly attached to the surface of the cylindrical body 221 away from the conveying pipe 222, and these meshing teeth mesh with a drive gear 24. A through hole is provided at the upper end of the fixing frame 23, and the roller 22 is rotatably placed in the through hole. In use, a drive motor is placed at the bottom of the fixing frame 23. The output shaft of the drive motor is fixedly connected to the drive gear 24. When the drive motor rotates, it drives the drive gear 24 to rotate. The drive gear 24 meshes with the meshing teeth on the roller 22, thereby driving the roller 22 to rotate, thus achieving a stable rolling state for the roller 22. Figure 9 As shown, different mixed solutions are placed in the first delivery chamber 224 and the second delivery chamber 225. For example, sodium alginate is placed in one chamber and cell suspension is placed in the other chamber. Sodium alginate and cell suspension are mixed simultaneously from the delivery section to the delivery pipe 222 and then sprayed out from the L-shaped nozzle.

[0022] The number of devices and processing scale described herein are for simplification of the invention. Applications, modifications, and variations of this invention will be readily apparent to those skilled in the art. Although embodiments of the invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. It can be applied to various fields suitable for this invention, and further modifications can be readily implemented by those skilled in the art. Therefore, without departing from the general concept defined by the claims and their equivalents, this invention is not limited to the specific details and illustrations shown and described herein.

Claims

1. A device for preparing cell microspheres, characterized in that, include: The injection pump assembly (10), the mixing assembly (20) and the injection assembly (30) are disposed adjacent to the injection pump assembly (10); The injection pump assembly (10) is used to provide power to expel the mixture in the mixing assembly (20) from the injection assembly (30); The mixing assembly (20) includes a fixed frame (23), a roller (22) rotatably connected to the fixed frame (23), a piston rod (21) sliding in the roller (22), and a drive gear (24) meshing with the roller (22); at least one piston rod (21) is provided in the roller (22).

2. The cell microsphere preparation apparatus as described in claim 1, characterized in that, The roller (22) includes a cylinder (221), one end of which is integrally connected to a conveying pipe (222), and a conveying chamber (223) is provided inside the cylinder (221), which is connected to the conveying pipe (222).

3. The cell microsphere preparation apparatus as described in claim 2, characterized in that, A ring of meshing teeth is fixed to the surface of the cylinder (221) away from the conveying pipe (222), and the meshing teeth mesh with the drive gear (24).

4. The cell microsphere preparation apparatus as described in claim 2, characterized in that, The cylindrical body (221) has a conical structure.

5. The cell microsphere preparation apparatus as described in claim 2, characterized in that, The conveying chamber (223) is cylindrical, and the angle between the axis of the conveying chamber (223) and the center line of the cone is between 5° and 30°.

6. The cell microsphere preparation apparatus as described in claim 2, characterized in that, The injection assembly (30) includes an L-shaped nozzle (31) which is internally connected to the delivery pipe (222).

7. The cell microsphere preparation apparatus as described in claim 1, characterized in that, The injection pump assembly (10) includes a slide rail (11), a slider (12) slidably disposed on the slide rail (11), and a disc (13) fixedly connected to one end of the slider (12). A piston rod (21) is abutted and connected on the disc (13).

8. The cell microsphere preparation apparatus as described in claim 7, characterized in that, The piston rod (21) is spherical at one end near the disk (13). The spherical end of the piston rod (21) rotates on the surface of the disk (13). The piston rod (21) is inserted into the delivery chamber (223) at one end of the disk (13).

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