Horizontal shaft stirring reactor for preparing mesenchymal stem cell spheres

Abstract

The invention discloses a horizontal shaft stirring reactor for preparing mesenchymal stem cell spheres, and relates to the technical field of mesenchymal stem cell sphere preparation, the horizontal shaft stirring reactor comprises a container used for containing a culture solution for stem cell balling and a stirring device arranged in the container, the top of the container is provided with an opening, and the top of the container is fixedly provided with a top cover capable of opening the opening; the stirring device comprises a stirring shaft which is transversely arranged and a plurality of stirring blades which are transversely arranged around the stirring shaft and are arranged in the container, the stirring shaft is driven by a driving assembly which is arranged outside the container to rotate, the driving assembly drives the stirring shaft to rotate in a magnetic coupling mode, and the driving shaft drives the stirring blades to stir the culture solution in the container; the balling of the mesenchymal stem cells is promoted. The horizontal stirring structure is innovated, the flow field distribution of the reactor is optimized, the problem of non-uniform shearing force in the cell sphere culture process is effectively solved, and a key technical support is provided for industrial production of high-quality stem cell spheres.

Description

Technical Field

[0001] The present invention relates to the technical field of mesenchymal stem cell sphere preparation, and specifically to a horizontal-axis stirring reactor for mesenchymal stem cell sphere preparation. Background Art

[0002] In the field of modern biomedicine, mesenchymal stem cells (MSCs) have shown great application value in tissue engineering, regenerative medicine, and cell therapy due to their multi-directional differentiation potential and immunomodulatory functions. Stem cell sphere culture technology can effectively maintain stem cell stemness and promote cell-to-cell signal transmission, and is a key means for large-scale production of high-quality stem cells. Currently, stem cell sphere preparation technology based on bioreactors has become a research hotspot, but existing devices still have significant deficiencies in aspects such as cell aggregation efficiency, microenvironment regulation, and engineering scale-up.

[0003] Traditional stirred bioreactors mostly adopt a vertical-axis stirring structure (such as propeller-type and turbine-type agitators). The shear force generated by them is unevenly distributed, which easily leads to damage to the cells on the surface of the stem cell sphere or fragmentation of the sphere. Especially during high-cell-density culture, the problem of cell apoptosis caused by shear force is more prominent. At the same time, the eddy current formed by vertical stirring easily causes mass transfer gradients of nutrients, oxygen, and signaling molecules in the culture system, resulting in uneven sizes of stem cell spheres and core necrosis, etc., seriously affecting the uniformity of the quality of cell spheres. Summary of the Invention

[0004] The purpose of the present invention is to provide a horizontal-axis stirring reactor for mesenchymal stem cell sphere preparation to solve the problems raised in the above background art.

[0005] To achieve the above purpose, the present invention provides the following technical solutions:

[0006] A horizontal-axis stirring reactor for mesenchymal stem cell sphere preparation includes a container for holding a culture solution for stem cell spheroid formation and a stirring device arranged in the container. The top of the container is an opening, and a top cover that can open the opening is fixed to the top of the container. The stirring device includes a horizontally arranged stirring shaft and a plurality of stirring blades arranged horizontally around the stirring shaft in the container. The stirring shaft is driven to rotate by a driving component arranged outside the container, and the driving component drives the stirring shaft to rotate through a magnetic coupling method. The stirring shaft drives the stirring blades to stir the culture solution in the container to promote mesenchymal stem cell spheroid formation.

[0007] Compared with the prior art, the beneficial effects of the present invention are:

[0008] By innovating the horizontal stirring structure and optimizing the flow field distribution of the reactor, the present invention effectively solves the problem of uneven shear force during the cell sphere culture process, providing key technical support for the industrial production of high-quality stem cell spheres. Description of the Drawings

[0009] Figure 1 It is a schematic structural diagram of a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres.

[0010] Figure 2 It is an exploded view of a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres.

[0011] Figure 3 It is a schematic structural diagram of the stirring device in a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres.

[0012] Figure 4 It is a schematic structural diagram of the mounting bracket in a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres.

[0013] Figure 5 It is a schematic structural diagram of the active magnetic ring and the protective cover in a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres.

[0014] Figure 6 It is a schematic structural diagram of the interior of the container in a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres. Detailed implementation manners

[0015] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0016] Please refer to Figures 1 to 6 , in the embodiments of the present invention, a horizontal-axis stirring reactor for preparing mesenchymal stem cell spheres includes a container 1 for containing a culture solution for stem cell spheroid formation and a stirring device 5 disposed in the container 1. The top of the container 1 is an opening, and a top cover 2 that can open the opening is fixed to the top of the container 1. The stirring device 5 includes a horizontally disposed stirring shaft 51 and a plurality of stirring blades 52 disposed in the container 1 and horizontally arranged around the stirring shaft 51. The stirring shaft 51 is driven to rotate by a driving assembly 3 disposed outside the container 1. The driving assembly 3 drives the stirring shaft 51 to rotate by means of magnetic coupling. The stirring shaft 51 drives the stirring blades 52 to stir the culture solution in the container 1 to promote the formation of mesenchymal stem cell spheres.

[0017] By innovating the horizontal stirring structure and optimizing the flow field distribution of the reactor, the present invention effectively solves the problem of uneven shear force during the cell sphere culture process, providing key technical support for the industrial production of high-quality stem cell spheres; and by setting the stirring method of the horizontal stirring shaft structure, a series of problems caused by the vortex formed by vertical stirring in the prior art are avoided.

[0018] In addition, the present invention drives the stirring device 5 by means of magnetic coupling. Both sides of the entire container 1 are completely sealed without any openings, effectively avoiding the problem of leakage easily caused by the transmission shaft penetrating the container wall in the prior art.

[0019] The stirring blades 52 are distributed in a plurality of circumferences with different diameters around the stirring shaft 51. In order to improve the uniformity during the stirring process, the stirring blades 52 in the same radial direction are evenly distributed. Specifically, when setting, the stirring blades are horizontally arranged rods, and both ends of the rods are connected to a plurality of connecting rods arranged radially along the stirring shaft 51, and the ends of the connecting rods are fixedly connected to the stirring shaft 51.

[0020] The stirring device 5 further includes a driven magnetic ring 53 coaxially arranged with the stirring shaft 51. The driven magnetic ring 53 rotates synchronously with the stirring shaft 51 and the stirring blades 52. The driving assembly 3 includes a driving magnetic ring 31 arranged outside the container 1 and a driving motor 33 arranged outside the container 1. The driving motor 33 drives the driving magnetic ring 31 to rotate, and the driving magnetic ring 31 drives the driven magnetic ring 53 to rotate across the side wall of the container 1, thereby driving the stirring shaft 51 and the stirring blades 52 to stir the culture solution in the container 1.

[0021] The magnetic poles of the driven magnetic ring 53 and the driving magnetic ring 31 are opposite. When the driving magnetic ring 31 rotates, it can drive the driven magnetic ring 53 to rotate, realizing the transmission of power.

[0022] An inner ring 311 coaxially arranged is fixedly installed inside the driving magnetic ring 31 through a connecting rod. Outside the container 1, there is also a protective cover 32 covering the driving magnetic ring 31 on the outer wall of the container 1. A driving shaft 321 coaxially arranged with the protective cover 32 is rotatably connected to the protective cover 32. The driving shaft 321 coaxially passes through the inside of the inner ring 311 and is fixedly connected to the inner ring 311. The driving shaft 321 is fixedly connected to the driving shaft of the driving motor 33. The driving motor 33 drives the driving magnetic ring 31 to rotate through the driving shaft 321 and the inner ring 311.

[0023] During specific implementation, a pin can penetrate the inner ring 311 and the driving shaft 321 to make the two rotate synchronously, and the driving shaft 321 is synchronously rotated with the output shaft of the driving motor 33 by means of key connection.

[0024] An installation ring 322 for fixedly connecting with the container 1 is fixed on the outer edge of the protective cover 32.

[0025] During specific installation, a plurality of studs passing through the installation ring 322 are fixed on the outer wall of the container 1. Nuts are threadedly connected to the studs, and the installation ring 322 is pressed against the outer wall of the container 1 by the nuts for fixation, ensuring the sealing performance of the container 1.

[0026] The stirring device 5 is installed in the container 1 through two mounting brackets 4 symmetrically arranged with respect to the stirring device 5.

[0027] The mounting bracket 4 includes an n-shaped bracket arranged vertically. On the inner walls of two sides of the container 1 perpendicular to the stirring shaft 51 of the stirring device 5, mounting grooves 11 with only the top being open are fixed. One vertical leg of the n-shaped bracket is inserted into the mounting groove 11 from top to bottom. A fixed shaft 42 that can be inserted into the stirring shaft 51 is fixed on the outer side of the other vertical leg of the n-shaped bracket. The stirring shaft 51 is installed by inserting the fixed shafts 42 of the mounting brackets 4 on both sides of the stirring device 5 into the stirring shaft 51 from both ends of the stirring shaft 51.

[0028] The distance between the two legs of the n-shaped bracket is the same as the wall thickness of the mounting groove 11 located inside the n-shaped bracket.

[0029] In order to facilitate inserting the mounting bracket 4 into the mounting groove 11, a guiding inclined surface 12 that opens outward may be provided at the notch of the top of the mounting groove 11 to reduce the installation difficulty of the mounting bracket 4.

[0030] The top of the mounting bracket 4 is flush with the bottom surface of the top cover 2 fixed to the top of the container 1. When the top cover 2 is fixed to the top of the container 1, the mounting bracket 4 is pressed and fixed in the mounting groove 11. After the stirring device 5 is installed, it can be ensured that the mounting bracket 4 is stable and does not shake, improving the stability of the installation of the stirring device 5.

[0031] It should be noted that the drive motor 33 in the present invention can select a three-phase vertical motor with the model 80M1-4 and is powered by the municipal power supply system. Data such as the rotation speed and direction during its working process can be controlled by a dedicated controller. This technology is a mature existing technology and will not be elaborated here. In addition, in order to ensure that the rotation speed of the stirring device 5 in the container 1 is appropriate, a reduction motor can also be provided between the drive motor and the drive shaft 321; this technology is easily conceivable by professionals in this technical field and will not be elaborated in this application.

[0032] Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention, and the content not described in detail in this specification belongs to the well-known existing technology of professionals in this field.

Claims

1. A horizontal axis stirred reactor for preparing mesenchymal stem cell spheres, characterized in that: The invention comprises a container (1) for containing culture fluid for stem cell sphering and a stirring device (5) arranged in the container (1); the top of the container (1) is set to be open, and a top cover (2) that can be opened is fixed on the top of the container (1); the stirring device (5) comprises a stirring shaft (51) arranged transversely and a plurality of stirring blades (52) arranged transversely around the stirring shaft (51) arranged in the container (1); the stirring shaft (51) is driven to rotate by a driving component (3) arranged outside the container (1); the driving component (3) drives the stirring shaft (51) to rotate by means of magnetic coupling; the stirring shaft (51) drives the stirring blades (52) to stir the culture fluid in the container (1), thereby promoting the sphering of mesenchymal stem cells.

2. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 1, characterized in that: The stirring blades (52) are distributed on a plurality of concentric circles with different diameters around the stirring shaft (51).

3. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 1, characterized in that: The stirring device (5) further comprises a driven magnetic ring (53) coaxially arranged with the stirring shaft (51), the driven magnetic ring (53) rotates synchronously with the stirring shaft (51) and the stirring blade (52), the driving assembly (3) comprises an active magnetic ring (31) arranged outside the container (1) and a driving motor (33) arranged outside the container (1), the driving motor (33) drives the active magnetic ring (31) to rotate, and the active magnetic ring (31) drives the driven magnetic ring (53) to rotate in a magnetic coupling manner through the side wall of the container (1), thereby driving the stirring shaft (51) and the stirring blade (52) to stir the culture solution in the container (1).

4. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 3, characterized in that: The active magnetic ring (31) is fixed with a coaxially arranged inner ring (311) via a connecting rod. The container (1) is also provided with a protective cover (32) covering the active magnetic ring (31) on the outer wall of the container (1). The protective cover (32) is rotatably connected with a drive shaft (321) coaxially arranged with the protective cover (32). The drive shaft (321) coaxially passes through the inner ring (311) and is fixedly connected to the inner ring (311). The drive shaft (321) is fixedly connected to the drive shaft of a drive motor (33). The drive motor (33) drives the active magnetic ring (31) to rotate via the drive shaft (321) and the inner ring (311).

5. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 4, characterized in that: A mounting ring (322) is fixed to the outer edge of the protective cover (32) for fixed connection with the container (1).

6. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 5, characterized in that: A plurality of studs passing through the mounting ring (322) are fixed on the outer wall of the container (1), nuts are threadedly connected to the studs, and the mounting ring (322) is fixed to the outer wall of the container (1) via the nuts.

7. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 1, characterized in that: The stirring device (5) is installed in the container (1) via two mounting brackets (4) that are symmetrically arranged with respect to the stirring device (5).

8. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 7, characterized in that: The mounting bracket (4) comprises an n-shaped bracket arranged vertically, and mounting grooves (11) with only the top being open are fixed on the inner walls on both sides of the container (1) perpendicular to the stirring shaft (51) of the stirring device (5), and one vertical leg of the n-shaped bracket is inserted into the mounting groove (11) from top to bottom, and a fixed shaft (42) that can be inserted into the stirring shaft (51) is fixed on the outer side of the other vertical leg of the n-shaped bracket, and the fixed shafts (42) of the mounting brackets (4) on both sides of the stirring device (5) are inserted into the stirring shaft (51) from both ends to achieve the installation of the stirring shaft (51).

9. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 8, characterized in that: The distance between the two legs of the n-shaped bracket is the same as the wall thickness of the mounting groove (11) located inside the n-shaped bracket.

10. The horizontal axis stirred reactor for preparing mesenchymal stem cell spheres according to claim 8, characterized in that: The top of the mounting bracket (4) is flush with the bottom surface of the top cover (2) fixed to the top of the container (1); when the top cover (2) is fixed to the top of the container (1), the mounting bracket (4) is pressed into the mounting groove (11) and fixed.

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