Stirring wheel and bioreactor

Abstract

A stirring wheel and a bioreactor. The stirring wheel comprises a hub, a plurality of blades and a paddle group, wherein the hub is configured as a hollow cylinder, and the hub has openings on two opposite ends thereof, and is provided with at least two holes spaced apart in the circumferential direction thereof; the plurality of blades are distributed spaced apart on the outer peripheral wall of the hub; and the paddle group is mounted inside the hub and configured to enable a fluid inside the hub to flow in the axial direction of the hub.

Description

Stirring wheel and bioreactor

[0001] Related applications

[0002] This application claims priority to Chinese patent application No. 202311285092.8 filed on September 28, 2023, the entire contents of which are incorporated by reference into this application. Technical Field

[0003] The present application relates to the technical field of stirring wheels, and in particular to a stirring wheel and a bioreactor. Background Art

[0004] Typically, acclimated adherent animal cells or suspension cells experience a tendency to aggregate and grow continuously during long-term culture processes without disrupting the aggregation. Throughout the culture cycle, the stirring speed must be adjusted in real time based on the cell aggregation characteristics. This adjustment also requires ensuring that extremely low shear forces are maintained throughout the process to prevent excessive shear forces from affecting cell activity, disrupting cell aggregation, and ultimately, proliferation.

[0005] At present, most of the lifting agitator paddles are used to culture carrier-free suspended cells or cell spheres. This type of agitator paddle is composed of multiple smaller elephant ear paddles, and needs to be at a relatively high stirring speed to provide a better flow field to meet the requirements of cell suspension culture. However, this type of lifting agitator paddle can only be used in bioreactors with a smaller working volume (culture fluid volume), and cannot meet the requirements of bioreactors with larger working volumes for flow field mixing and cell agitation. If the speed of the agitator paddle is increased too much, the shear force will be amplified exponentially, which cannot meet the extremely low shear force requirements of suspended cell sphere culture, and may result in poor cell culture proliferation effect and failure to achieve the expected culture.

[0006] Summary of the Invention

[0007] The main purpose of this application is to propose a stirring wheel to improve the mixing uniformity of the flow field inside the bioreactor.

[0008] To achieve the above objectives, the bioreactor proposed in this application includes:

[0009] The wheel hub is in a hollow cylindrical shape, with openings at opposite ends of the wheel hub, and at least two openings are spaced apart in the circumferential direction of the wheel hub;

[0010] a plurality of blades, each of which is connected to the hub and spaced apart on an outer peripheral wall of the hub; and

[0011] A blade group is installed inside the hub and is used to make the fluid inside the hub flow in the axial direction of the hub.

[0012] In one embodiment, the plurality of blades extend in the axial direction of the hub, the blades include connected straight sections and bent sections, the bent sections are located at the ends of the blades, and the plurality of bent sections are bent in the same direction along the circumference of the hub.

[0013] In one embodiment, the opening is located between any two adjacent blades.

[0014] In one embodiment, the number of the openings is set to two, and the two openings are symmetrically distributed on the hub.

[0015] In one embodiment, the number of the openings is set to two, and the two openings are staggered and distributed on the hub.

[0016] In one embodiment, an angle formed by a line connecting the two openings and the hub axis is greater than or equal to 90°.

[0017] In one embodiment, the opening extends in the axial direction of the hub.

[0018] In one embodiment, the length of the opening is set to L, the axial length of the hub is set to H, and L / H satisfies the range of 1 / 2 to 1.

[0019] In one embodiment, the opening includes a plurality of liquid passage openings, and the plurality of liquid passage openings are distributed in an axial direction of the hub at intervals.

[0020] In one embodiment, the length of the blade is consistent with the axial length of the hub.

[0021] In one embodiment, the number of the blades is set to 4 to 20.

[0022] In one embodiment, the number of the blades is set to 8 to 16.

[0023] In one embodiment, the blade group includes an upper blade group and a lower blade group, and the upper blade group and the lower blade group are spaced apart in the axial direction of the hub.

[0024] In one embodiment, the upper blade group includes two first blades, which are connected to the inner circumferential wall of the hub, and the two first blades are spaced apart in the circumferential direction of the inner circumferential wall of the hub; the lower blade group includes two second blades, which are connected to the inner circumferential wall of the hub, and the two second blades are spaced apart in the circumferential direction of the inner circumferential wall of the hub.

[0025] The present application also proposes a bioreactor, comprising a bioreactor stirring wheel, which includes a hub, multiple blades and a blade group. The hub is arranged in a hollow cylindrical shape, has openings at opposite ends of the hub, and the hub is provided with at least two openings at intervals in its circumferential direction; the multiple blades are respectively connected to the hub, and the multiple blades are distributed at intervals on the outer peripheral wall of the hub; the blade group is installed inside the hub, and the blade group is used to make the fluid inside the hub flow in the axial direction of the hub.

[0026] The technical solution of the present application stirs the culture medium in the bioreactor by adopting a stirring wheel with multiple blades circumferentially spaced on the outer wall of the hub. It can improve the mixing uniformity of the flow field and the amount of cell tumbling while reducing the shear force of the stirring wheel on the cells. An opening is opened on the hub, and a blade group is arranged inside the hub so that the culture medium inside the hub can be mixed with the culture medium outside the hub. The stirring wheel in the present application still has a good stirring effect in a bioreactor with a larger working volume. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying any creative work.

[0028] FIG1 is a velocity cloud diagram of a bioreactor stirred with an elephant ear paddle in the background art (the working volume of the bioreactor is 500 ml);

[0029] FIG2 is a velocity cloud diagram of a bioreactor stirred with an elephant ear paddle in the background art (the bioreactor working volume is 5 L);

[0030] FIG3 is a velocity cloud diagram of a bioreactor stirred with an elephant ear paddle in the background art (the bioreactor working volume is 10 L);

[0031] FIG4 is a schematic structural diagram of a stirring wheel according to an embodiment;

[0032] FIG5 is a schematic structural diagram of FIG4 from another perspective;

[0033] FIG6 is a schematic structural diagram of a stirring wheel in a cross-sectional view;

[0034] FIG7 is a schematic structural diagram of a bioreactor from a cross-sectional perspective;

[0035] FIG8 is a schematic structural diagram of the stirring wheel from another cross-sectional perspective;

[0036] FIG9 is a shear force distribution diagram of a wheel hub without openings;

[0037] FIG10 is a diagram showing the shear force distribution when the hub has one opening;

[0038] FIG11 is a shear force distribution diagram of a wheel hub with two openings;

[0039] FIG12 is a diagram showing the shear force distribution of a wheel hub with four openings;

[0040] FIG13 is a velocity cloud diagram of a bioreactor with a working volume of 5 L viewed from the front;

[0041] FIG14 is a velocity cloud diagram of a bioreactor with a working volume of 10 L viewed from the front;

[0042] FIG15 is a velocity contour diagram of a bioreactor with a working volume of 5 L from a top view;

[0043] FIG16 is a velocity contour diagram of a bioreactor with a working volume of 10 L from a top view;

[0044] FIG17 is a velocity contour diagram of a bioreactor with a working volume of 5 L viewed from above;

[0045] FIG18 is a velocity contour diagram of a bioreactor with a working volume of 10 L viewed from above;

[0046] FIG19 is a velocity contour diagram of a bioreactor when two openings are symmetrically distributed;

[0047] FIG20 is a velocity contour diagram of a bioreactor when two openings are symmetrically distributed from above;

[0048] FIG21 is a velocity contour diagram of a bioreactor viewed from above when two openings are symmetrically distributed;

[0049] FIG22 is a velocity contour diagram of a bioreactor when two openings are staggered;

[0050] FIG23 is a velocity contour diagram of a bioreactor when two openings are staggered;

[0051] FIG24 is a velocity contour diagram of a bioreactor when two openings are staggered;

[0052] FIG25 is a velocity contour diagram of the bioreactor when the number of blades is 4;

[0053] FIG26 is a velocity contour diagram of the bioreactor when the number of blades is 6;

[0054] FIG27 is a velocity contour diagram of the bioreactor when the number of blades is 8;

[0055] FIG28 is a velocity contour diagram of the bioreactor when the number of blades is 12;

[0056] FIG29 is a velocity contour diagram of the bioreactor when the number of blades is 16;

[0057] FIG30 is a velocity contour diagram of the bioreactor when the number of blades is 20;

[0058] FIG31 is a velocity contour diagram of a bioreactor when the number of blades is 4, viewed from above;

[0059] FIG32 is a velocity contour diagram of the bioreactor when the number of blades is 6;

[0060] FIG33 is a velocity contour diagram of the bioreactor when the number of blades is 8;

[0061] FIG34 is a velocity contour diagram of the bioreactor when the number of blades is 12;

[0062] FIG35 is a velocity contour diagram of the bioreactor when the number of blades is 16;

[0063] FIG36 is a velocity contour diagram of the bioreactor when the number of blades is 20 as viewed from above.

[0064] Description of Figure Numbers:

[0065] The realization of the objectives, functional features and advantages of this application will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0066] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0067] It should be noted that if the embodiments of the present application involve directional indications (such as up, down, left, right, front, back, etc.), the directional indications are only used to explain the relative position relationship, movement status, etc. between the various components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indications will also change accordingly.

[0068] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present application, the descriptions of "first", "second", etc. are only for descriptive purposes and cannot be understood as indicating or suggesting their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of such features. In addition, if the meaning of "and / or" appearing in the full text is to include three parallel schemes, taking "A and / or B" as an example, it includes scheme A, or scheme B, or a scheme in which A and B are satisfied at the same time. In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the ability of ordinary technicians in this field to implement. When the combination of technical solutions is mutually contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0069] During long-term culture, adherent or suspended animal cells, which have been acclimated, continuously aggregate and grow without disrupting the aggregates. Throughout the culture cycle, the stirring speed must be adjusted in real time based on the cell aggregation characteristics. During this adjustment, the extremely low shear force requirement must be maintained throughout the process to avoid excessive shear forces that could affect cell activity, disrupt cell aggregation, and ultimately promote proliferation.

[0070] Currently, conventional lifting impellers are used to culture carrier-free suspended cells or cell spheroids. These impellers, composed of multiple small elephant-ear paddles, require relatively high stirring speeds to provide a good flow field, meeting the requirements of cell suspension culture. However, high speeds exponentially amplify shear forces, failing to meet the extremely low shear requirements of suspended cell spheroid culture. This can lead to poor cell proliferation and failure to achieve desired cell culture results.

[0071] Please refer to Figures 1 to 3, which are flow field velocity simulations for a bioreactor with a lifting impeller. It should be noted that the left side of the simulation shows the numerical change in flow velocity. In the original image, the color changes from blue to yellow to red as the flow velocity increases from low to high (i.e., from bottom to top in the image). After grayscale processing of the simulation, the color changes from dark to light to dark as the flow velocity increases from low to high. The flow velocity and color correspond to the color regions within the bioreactor on the right side of the simulation. Generally speaking, areas near the impeller are lighter, indicating higher flow velocity, while areas farther from the impeller are darker, indicating lower flow velocity.

[0072] Figure 1 is a flow velocity simulation diagram for a bioreactor with a working volume of 500 ml. It can be seen that the flow velocity around the impeller is high, the flow velocity distribution is uniform, and the flow field mixing is good. Figures 2 and 3 are flow velocity simulation diagrams for bioreactors with working volumes of 5 L and 10 L, respectively. It can be clearly seen that compared to Figure 1, the flow velocity around the impeller is lower, the flow velocity distribution is uneven, and the flow field mixing is poor. Therefore, it can be concluded that this type of lifting impeller is not suitable for bioreactors with larger working volumes.

[0073] In view of this, the present application proposes a stirring wheel 10 .

[0074] In an embodiment of the present application, as shown in Figures 4 to 8, the stirring wheel 10 includes a hub 100, a plurality of blades 200 and a blade group 300. The hub 100 is arranged in a hollow cylindrical shape, and has openings 110 at opposite ends of the hub 100. The hub 100 is provided with at least two openings 120 at intervals in its circumferential direction; the plurality of blades 200 are respectively connected to the hub 100, and the plurality of blades 200 are distributed at intervals on the outer peripheral wall of the hub 100; the blade group 300 is installed inside the hub 100, and the blade group 300 is used to make the fluid inside the hub 100 flow in the axial direction of the hub 100.

[0075] The stirring wheel 10 in this embodiment is used in a bioreactor to stir the culture medium and cells in the bioreactor so that the adherent animal cells or suspended cells can obtain the required gas and culture medium. The stirring wheel 10 of the present application is generally set to a speed between 5 rpm and 50 rpm when culturing adherent animal cells or suspended cells. At this speed, the various culture media in the bioreactor can be fully mixed and the basic amount of tumbling required by the cells can be met. When the stirring wheel 10 is within this speed range, the average shear force is between 0.03 Pa and 0.2 Pa, which meets the low shear force requirements of the adherent animal cells or suspended cells.

[0076] The hub 100 is shaped like a hollow cylinder, and both ends of the stirring wheel 10 have openings 110, so that the culture medium can flow from the inside of the hub 100. The stirring wheel 10 is provided with a plurality of blades 200 connected to the hub 100, and the plurality of blades 200 extend in the axial direction of the hub 100. Considering that the blades 200 on the hub 100 must have sufficient stirring force, the number of blades 200 cannot be too small, and the number of blades 200 must be at least greater than or equal to four. In this way, the radial length of the hub 100 cannot be set too small to ensure that there is enough space to install multiple blades 200. The shape of the blades 200 can be a straight plate, an arc plate, or a special shape, which is not specifically limited here.

[0077] In order to ensure that the culture medium inside the hub 100 can be fully mixed and stirred, the stirring wheel 10 also includes a blade group 300. The blade group 300 is arranged inside the hub 100. The blade group 300 can make the culture medium inside the hub 100 flow in the axial direction of the hub 100. In order to make the flow effect of the culture medium inside the hub 100 better, the blade group 300 can be provided with multiple blades, wherein the multiple blades can be arranged along the circumferential direction of the hub 100, the multiple blades can also be arranged along the axial direction of the hub 100, and the multiple blades can also be arranged both along the circumferential direction of the hub 100 and along the axial direction of the hub 100. The arrangement of the blades in the blade group 300 is not specifically limited here.

[0078] In one embodiment, the flow of the culture medium inside the hub 100 is balanced with the flow of the culture medium outside the hub 100 so that the two parts of the culture medium are mixed more evenly. At least two openings 120 are opened on the inner wall of the hub 100. It should be noted that, referring to Figures 9 to 12, it should be noted that Figures 9 to 12 are shear force distribution diagrams of the bioreactor. The original shear force changes from low to high (i.e., from bottom to top in the figure) with a color gradient from blue to yellow and then to red. After grayscale processing of the simulation diagram, the shear force changes from low to high to a color gradient from dark to light and then to dark. The shear force and color correspond to the color areas in the simulation diagram and the right bioreactor. Generally speaking, the area close to the stirring paddle is lighter in color, indicating higher shear force, and the area away from the stirring paddle is darker in color, indicating lower shear force.

[0079] Figure 9 shows a comparative solution without openings 120 on the hub 100, Figure 10 shows a comparative solution with one opening 120 on the hub 100, and Figure 11 shows a solution with two openings 120 on the hub 100 of this embodiment. Figure 12 shows a solution with two openings 120 on the hub 100. As can be clearly seen in Figure 9 , the interior of the hub 100 is very dark, indicating very low shear forces within the hub 100. In Figure 10 , the interior of the hub 100 is also darker, indicating uneven shear force distribution within the hub 100. Therefore, the comparative solutions of Figures 9 and 10 may result in poor tumbling of adherent or suspended cells within the hub 100. The interior of the hub 100 in Figure 11 is much brighter than in Figures 9 and 10 , indicating higher shear forces within the hub 100. Therefore, the hub 100 should have at least two openings 120.

[0080] Please refer to Figures 13 to 18 , where Figures 13 , 15 , and 17 are flow velocity cloud maps for this embodiment at a 5 L working volume, and Figures 14 , 16 , and 18 are flow velocity cloud maps for this embodiment at a 10 L working volume. It should be noted that the left side of the simulation graph shows the numerical change in flow velocity. In the original graph, the flow velocity changes from low to high (i.e., from bottom to top in the graph) with a gradual color change from blue to yellow to red. After grayscale processing of the simulation graph, the flow velocity changes from low to high with a gradual color change from dark to light to dark again. The color of the flow velocity corresponds to the color region within the bioreactor on the right side of the simulation graph. Generally speaking, areas near the impeller 10 and blades 200 are lighter in color, indicating higher flow velocity, while areas farther from the impeller 10 are darker, indicating lower flow velocity. The flow velocity inside the impeller 10 is lower than that outside the impeller 10. If a dark region appears in the middle of the light region surrounding the impeller 10, it indicates that the flow velocity there is higher than the surrounding light region. The description here is also applicable to the flow velocity cloud diagrams in the following embodiments, so it is only described here once.

[0081] It can be seen that by comparing Figures 13 and 14 with Figures 2 and 3, the stirring wheel of this embodiment has good flow field mixing uniformity compared to the lifting stirring paddle at a large working volume. Therefore, it is better to choose the stirring wheel of this embodiment at a large working volume.

[0082] In one embodiment, combined with the simulation data provided in the table below, the rotation speed of the lifting agitator is 60 rpm, the agitator is divided into an upper agitator and a lower agitator, the average shear rate around the upper agitator is 12.396, and the average shear rate around the lower agitator is 12.388. There is a point with maximum shear force around the upper agitator and the lower agitator, where the maximum shear rate is 224.7. The average shear rate of other areas in the tank except the upper agitator and the lower agitator is 3.137.

[0083] Among them, the bioreactor in Figure 2 is a flow field velocity simulation diagram when the working volume is 5L. It can be seen that the flow field velocity of the stirring paddle bioreactor is relatively high at this time, and the flow field mixing uniformity is better. Combined with the simulation data provided in the table below, the rotation speed of the lifting stirring paddle is 60rpm at this time, and the stirring paddle is divided into an upper stirring paddle and a lower stirring paddle. The average shear rate around the upper stirring paddle is 12.289, and the average shear rate around the lower stirring paddle is 14.210. There is a point with maximum shear force around the upper stirring paddle and the lower stirring paddle, where the maximum shear rate is 631.4. The average shear rate of other areas in the tank body except the upper stirring paddle and the lower stirring paddle is 2.128. In the table, the average shear force around the multiple blades of the stirring wheel of this embodiment is 4.098, the average shear force in other areas is 2.781, and the shear force at the maximum point is 99.41. It can be concluded that the average shear force of the stirring wheel 10 of this embodiment around the multiple blades 200 of the stirring wheel 10 is smaller than the average shear force of the lifting stirring paddle around the stirring blades, and the shear force at the maximum point of the stirring wheel 10 is much smaller than the shear force at the maximum point of the lifting stirring paddle. In view of this, the low shear force of the stirring wheel 10 of this embodiment is more suitable for the culture environment required for adherent cells or suspended cells.

[0084] The technical solution of the present application stirs the culture medium in the bioreactor by adopting a stirring wheel 10 with multiple blades 200 circumferentially spaced on the outer wall of the hub 100. This can improve the mixing uniformity of the flow field and the amount of cell tumbling while reducing the shear force of the stirring wheel 10 on the cells. An opening 120 is provided on the hub 100, and a blade group 300 is provided inside the hub 100 so that the culture medium inside the hub 100 can be mixed with the culture medium outside the hub 100. In addition, the stirring wheel 10 in the present application still has a good stirring effect in a bioreactor with a larger working volume.

[0085] In one embodiment, referring to Figures 5 to 7, a plurality of blades 200 extend in the axial direction of the hub 100, and the blades 200 include connected straight plate sections 210 and bent sections 220, wherein the bent sections 220 are located at the ends of the blades 200, and the bent sections 220 are bent along the circumference of the hub 100.

[0086] Considering that if the blade 200 is set as a straight plate as a whole, the cells will be directly dispersed, and the adherent cells cannot be cultured well. Furthermore, the blade 200 includes a straight plate section 210 and a bent section 220 connected to each other, wherein the bent section 220 is located at the end of the blade 200, and the bent section 220 is bent along the circumference of the hub 100. The setting of the straight plate section 210 can expand the stirring range of the blade 200 and fully stir the culture medium in the bioreactor. During the rotation of the stirring wheel 10, the setting of the bent section 220 will make it easier for the stirring wheel 10 to drive a part of the adherent cells or suspended cells to rotate together, so that the cultured cells can be fully mixed with the culture medium and absorb the nutrients and required gases in the culture medium. Moreover, the bent section 220 is bent along the circumference of the hub 100, so that the bending direction of the bent section 220 is consistent with the rotation trajectory of the stirring wheel 10, which is easier to drive the cultured cells to rotate together.

[0087] In one embodiment, referring to FIG. 4 to FIG. 8 , the opening 120 is located between any two adjacent blades 200 .

[0088] It should be noted that, in order to facilitate the opening of the opening 120, the opening 120 is opened between any two adjacent blades 200, and correspondingly, other openings 120 are also opened between any two adjacent blades 200, wherein two or more openings 120 can be located between the same two adjacent blades 200, or between two different adjacent blades 200. Taking into account that multiple openings 120 are spaced apart in the circumferential direction, opening multiple openings 120 between two different adjacent blades 200 is an optional embodiment.

[0089] In one embodiment, referring to Figures 5, 19, 20, and 21, Figures 19 to 21 illustrate flow field velocity distribution diagrams when the openings 120 are symmetrically arranged. Figure 19 is a velocity cloud diagram from a side cross-sectional perspective, Figure 20 is a velocity cloud diagram from an upper side perspective, and Figure 21 is a velocity cloud diagram from a lower side perspective. The number of openings 120 is set to two, and the two openings 120 are symmetrically distributed on the hub 100.

[0090] When two openings 120 are provided, the culture medium flowing into the interior of the hub 100 from one opening 120 can flow out from the other opening 120, that is, the culture medium inside the hub 100 can flow not only in the axial direction but also in the radial direction of the hub 100, thereby enhancing the mixing degree between the culture medium in the internal flow channel of the hub 100 and the culture medium outside the hub 100, and also making the velocity distribution of the flow field inside the hub 100 more uniform. It can be seen from Figures 19 to 21 that the flow velocity distribution inside and outside the hub 100 is relatively uniform.

[0091] In one embodiment, referring to Figures 8, 22, 23, and 24, Figures 23 and 24 illustrate flow field velocity distribution diagrams when the openings 120 are arranged in pairs, Figure 22 is a velocity nephogram from a side cross-sectional perspective, Figure 23 is a velocity nephogram from an upper perspective, and Figure 24 is a velocity nephogram from a lower perspective. The number of the openings 120 is set to two, and the two openings 120 are staggered on the hub 100.

[0092] When two openings 120 are provided, the culture medium flowing into the interior of the hub 100 from one opening 120 can flow out from the other opening 120, that is, the culture medium inside the hub 100 can flow not only in the axial direction but also in the radial direction of the hub 100, thereby enhancing the mixing degree between the culture medium in the internal flow channel of the hub 100 and the culture medium outside the hub 100, and also making the velocity distribution of the flow field inside the hub 100 more uniform. It can be seen from Figures 19 to 21 that the flow velocity distribution inside and outside the hub 100 is relatively uniform.

[0093] In one embodiment, referring to FIG. 8 , the angle formed by the line connecting the two openings 120 and the axis of the hub 100 is greater than or equal to 90°.

[0094] Considering that the angle formed by the line connecting the two openings 120 and the axis of the hub 100 is less than 90°, after the culture medium flows into the interior of the hub 100 from one opening 120, it is not easy to flow out directly through the other opening 120, which may cause the flow velocity inside the hub 100 to slow down or the flow velocity to be uneven, thereby causing poor or uneven flipping, making it difficult to achieve the expected effect of culturing adherent cells or suspension cells.

[0095] In one embodiment, referring to FIG. 5 and FIG. 6 , the opening 120 extends in the axial direction of the hub 100 .

[0096] It should be noted that in order to further improve the fluidity of the culture medium between the outside and inside of the hub 100, the opening 120 is extended in the axial direction of the hub 100, that is, the length of the opening 120 in the axial direction of the hub 100 is longer, which can make the interaction of the culture medium between the opening 120 and the hub 100 better.

[0097] In one embodiment, referring to FIG. 5 , the length of the opening 120 is set to L, the axial length of the hub 100 is set to H, and L / H satisfies the range of 1 / 2 to 1.

[0098] The length of the opening 120 should satisfy a ratio to the axial length of the hub 100 of at least 1 / 2 to 1, that is, the length of the opening 120 can be greater than or equal to 1 / 2 of the axial length of the hub 100, and the length of the opening 120 must be less than the axial length of the hub 100. Within this range, the culture medium outside the hub 100 and the culture medium inside the hub 100 can circulate interactively, allowing the nutrients of the various components in the culture medium to circulate mutually.

[0099] In one embodiment, referring to FIG. 9 , the opening 120 includes a plurality of liquid passage openings, and the plurality of liquid passage openings are spaced apart and distributed in the axial direction of the hub 100 .

[0100] It should be noted that the plurality of openings 120 are not limited to being circumferentially spaced apart in the above embodiment. Furthermore, the openings 120 may include a plurality of liquid outlets, and the plurality of liquid outlets are spaced apart and arranged in the axial direction of the hub 100. Considering that a plurality of pipelines are inserted at different depths in the bioreactor, the main components of the nutrients entering each liquid outlet can be controlled so that the inflow components of different liquid outlets are different.

[0101] In one embodiment, referring to FIG. 4 , the bending directions of the plurality of bending segments 220 are consistent with the rotation direction of the hub 100 .

[0102] In the above embodiment, the bending section 220 is defined as being bent circumferentially. In order to further define the direction in which the multiple bending sections 220 bend circumferentially, the bending directions of the multiple bending sections 220 are consistent with the rotation direction of the hub 100. When the hub 100 rotates in a clockwise direction, the bending sections 220 bend in a clockwise direction. When the hub 100 rotates in a counterclockwise direction, the bending sections 220 rotate in a counterclockwise direction, so that the bending directions of the multiple bending sections 220 remain consistent, and the bending direction of the bending sections 220 is the same as the rotation direction, so that when the stirring wheel 10 is stirring, the bending portion can more easily drive the adherent cells or suspended cells in the bending portion to rotate together.

[0103] In one embodiment, referring to FIG. 5 and FIG. 6 , the length of the blade 200 is consistent with the axial length of the hub 100 .

[0104] In order to make the blade 200 have a larger rotation range, the length of the blade 200 is extended as much as possible. However, if the length of the blade 200 exceeds the axial length of the hub 100, the part of the blade 200 that exceeds the hub 100 is not connected to the hub 100, which may cause unstable rotation of the stirring wheel 10. Therefore, the length of the blade 200 is set to be consistent with the axial length of the hub 100. The length of the blade 200 can be the same as the axial length of the hub 100, or the length of the blade 200 can be slightly larger than the axial length of the hub 100, or the length of the blade 200 can be slightly smaller than the axial length of the hub 100.

[0105] In one embodiment, referring to FIG. 25 to FIG. 36 , the number of the blades 200 is set to 4 to 20.

[0106] 25 to 30 are velocity cloud diagrams of a bioreactor with an increasing number of bioreactor blades 200 from a frontal perspective, and FIG. 31 to 36 are velocity cloud diagrams of a bioreactor with an increasing number of bioreactor blades 200 from a top-down perspective.

[0107] It can be clearly seen from the front and top perspectives that when the number of blades 200 is 4 and 6, the flow velocity distribution around the blades 200 is significantly less uniform than when the number of blades 200 is 8 to 20. Furthermore, when the number of blades 200 is 12, the overall mixing of the flow field is the best.

[0108] Please refer to the table below, which shows the shear force data within the bioreactor when the number of blades 200 is 4, 6, 8, 12, 16, and 20. When the number of blades 200 is 4, the average shear force around the blades 200 is 3.998, the average shear force in other areas is 1.534, and the maximum shear force is 40.940. When the number of blades 200 is 6, the average shear force around the blades 200 is 4.276, the average shear force in other areas is 1.634, and the maximum shear force is 22.938. When the number of blades 200 is 8, the average shear force around the blades 200 is 4.416, the average shear force in other areas is 1.734, and the maximum shear force is 33.042. When the number of blades 200 is 12, the average shear force around the blades 200 is 4.622, the average shear force in other areas is 1.707, and the maximum shear force is 29.678. When there are 16 blades 200, the average shear force around the blades 200 is 4.721, the average shear force in other areas is 1.611, and the maximum shear force is 45.478. When there are 20 blades 200, the average shear force around the blades 200 is 4.510, the average shear force in other areas is 1.635, and the maximum shear force is 51.610.

[0109] As can be seen from the above table and data, when the number of blades 200 is 4 or 6, the average shear force around the blades 200 is too low, failing to achieve a satisfactory cell flipping effect. When the number of blades 200 is 16 or 20, the maximum shear force is too high, potentially inhibiting the growth and proliferation of adherent animal cells or suspended cells. When the number of blades 200 is 12, the shear force around the blades 200 is appropriate, and the maximum shear force is also relatively low. Therefore, setting the number of blades 200 to 12 is the optimal embodiment for this embodiment.

[0110] Referring to the influence of the number of blades 200 on the velocity distribution of the flow field in the bioreactor and the effect of the number of blades 200 on the shear force, it can be concluded that setting the number of blades 200 to 12 is an optional embodiment of this embodiment. Of course, setting the number of blades 200 to other numbers within the range of 4 to 20 can also meet the basic requirements for culturing adherent cells or suspended cells.

[0111] In one embodiment, referring to FIG. 25 to FIG. 36 , the number of the blades 200 is set to 8 to 16.

[0112] It should be noted that the number of blades 200 is set to 8 to 16.

[0113] In one embodiment, referring to the above analysis and table, culturing adherent animal cells and suspension cells requires as low a shear force as possible and better uniformity of flow rate. Thus, setting the number of blades 200 to 8 to 16 is a more preferred embodiment of the above embodiment.

[0114] In one embodiment, referring to FIG. 6 , the blade group 300 includes an upper blade group 310 and a lower blade group 320 . The upper blade group 310 and the lower blade group 320 are spaced apart in the axial direction of the hub 100 .

[0115] It should be noted that, considering that the hub 100 has a certain length in its axial direction, in order to further increase the flow rate of the culture medium inside the blade group 300 and the amount of cell tumbling, the blade group 300 is divided into an upper blade group 310 and a lower blade group 320. The upper blade group 310 and the lower blade group 320 respectively stir the culture medium inside the hub 100, thereby increasing the amount of cell tumbling inside the hub 100. The upper blade group 310 and the lower blade group 320 are spaced apart in the axial direction of the hub 100, and the upper blade group 310 and the lower blade group 320 are as close as possible to the two openings 110 of the hub 100, so that the upper blade group 310 and the lower blade group 320 stir and tumble the culture medium inside the hub 100 as much as possible.

[0116] In one embodiment, referring to Figure 6, the upper blade group 310 includes two first blades 311, which are connected to the inner circumferential wall of the hub 100, and the two first blades 311 are spaced apart in the circumferential direction of the inner circumferential wall of the hub 100; the lower blade group 320 includes two second blades 321, which are connected to the inner circumferential wall of the hub 100, and the two second blades 321 are spaced apart in the circumferential direction of the inner circumferential wall of the hub 100.

[0117] In the present embodiment, the upper blade group 310 includes two first blades 311, which is not limited to the present embodiment. Of course, the upper blade group 310 may also include three first blades 311, or four first blades 311 or even more first blades 311. The two first blades 311 are spaced apart on the circumference of the inner circumferential wall of the hub 100 to improve the stirring ability of the upper blade group 310. Correspondingly, in the present embodiment, the lower blade group 320 includes two second blades 321, which is not limited to the present embodiment. Of course, the lower blade group 320 may also include three second blades 321, or four second blades 321 or even more second blades 321. The two second blades 321 are spaced apart on the circumference of the inner circumferential wall of the hub 100 to improve the stirring ability of the lower blade group 320.

[0118] In addition, the upper blade group 310 and the lower blade group 320 are also connected to the inner wall of the hub 100, so that the upper blade group 310 and the multiple blades 200 outside the hub 100 can rotate together, which can be achieved through only one driving device, simplifying the driving method of the stirring wheel 10.

[0119] The present application also proposes a bioreactor, which includes a stirring wheel 10. The specific structure of the stirring wheel 10 refers to the above embodiment. Since the present bioreactor adopts all the technical solutions of all the above embodiments, it has at least all the beneficial effects brought by the technical solutions of the above embodiments, which will not be described one by one here.

[0120] The above description is merely an optional embodiment of the present application and does not limit the patent scope of the present application. All equivalent structural transformations made using the contents of the present application specification and drawings under the inventive concept of the present application, or direct / indirect application in other related technical fields are included in the patent protection scope of the present application.

Claims

1. A stirring wheel (10) for use in a bioreactor, wherein: The stirring wheel (10) comprises: A wheel hub (100), the wheel hub (100) being arranged in a hollow cylindrical shape, having openings (110) at opposite ends of the wheel hub (100), and the wheel hub (100) being arranged at least two openings (120) at intervals in a circumferential direction thereof; a plurality of blades (200), the plurality of blades (200) being respectively connected to the hub (100), and the plurality of blades (200) being distributed at intervals on the outer peripheral wall of the hub (100); and A blade group (300) is installed inside the wheel hub (100), and the blade group (300) is used to make the fluid inside the wheel hub (100) flow in the axial direction of the wheel hub (100).

2. The stirring wheel (10) according to claim 1, wherein: The plurality of blades (200) extend in the axial direction of the hub (100), and the blades (200) include connected straight plate sections (210) and bent sections (220), wherein the bent sections (220) are located at the ends of the blades (200), and the plurality of bent sections (220) are bent in the same direction along the circumference of the hub (100).

3. The stirring wheel (10) according to claim 2, wherein: The opening (120) is located between any two adjacent blades (200).

4. The stirring wheel (10) according to claim 3, wherein: The number of the openings (120) is set to two, and the two openings (120) are symmetrically distributed on the wheel hub (100).

5. The stirring wheel (10) according to claim 3, wherein: The number of the openings (120) is set to two, and the two openings (120) are staggered and distributed on the wheel hub (100).

6. The stirring wheel (10) according to claim 5, wherein: The angle formed by the line connecting the two openings (120) and the axis of the hub (100) is greater than or equal to 90°.

7. The stirring wheel (10) according to claim 3, wherein: The opening (120) extends in the axial direction of the hub (100).

8. The stirring wheel (10) according to claim 3, wherein: The length of the opening (120) is set to L, and the axial length of the hub (100) is set to H, and L / H satisfies the range of 1 / 2 to 1.

9. The stirring wheel (10) according to claim 3, wherein: The opening (120) includes a plurality of liquid passage openings, and the plurality of liquid passage openings are distributed at intervals in the axial direction of the hub (100).

10. The stirring wheel (10) according to claim 2, wherein: The length of the blade (200) is consistent with the axial length of the hub (100).

11. The stirring wheel (10) according to claim 2, wherein: The number of the blades (200) is set to be 4 to 20.

12. The stirring wheel (10) according to claim 11, wherein: The number of the blades (200) is set to be 8 to 16.

13. The stirring wheel (10) according to claim 2, wherein: The blade group (300) comprises an upper blade group (310) and a lower blade group (320), and the upper blade group (310) and the lower blade group (320) are spaced apart and distributed in the axial direction of the hub (100).

14. The stirring wheel (10) according to claim 13, wherein: The upper blade group (310) includes two first blades (311), the first blades (311) are connected to the inner circumferential wall of the hub (100), and the two first blades (311) are spaced apart in the circumferential direction of the inner circumferential wall of the hub (100); the lower blade group (320) includes two second blades (321), the second blades (321) are connected to the inner circumferential wall of the hub (100), and the two second blades (321) are spaced apart in the circumferential direction of the inner circumferential wall of the hub (100).

15. A bioreactor, wherein: The bioreactor comprises a stirring wheel (10) as claimed in any one of claims 1 to 14.

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