CN117431144A - Stirring wheel and bioreactor - Google Patents
Stirring wheel and bioreactor Download PDFInfo
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- CN117431144A CN117431144A CN202311285092.8A CN202311285092A CN117431144A CN 117431144 A CN117431144 A CN 117431144A CN 202311285092 A CN202311285092 A CN 202311285092A CN 117431144 A CN117431144 A CN 117431144A
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- 238000003756 stirring Methods 0.000 title claims abstract description 97
- 230000002093 peripheral effect Effects 0.000 claims abstract description 15
- 239000012530 fluid Substances 0.000 claims abstract description 5
- 230000004323 axial length Effects 0.000 claims description 15
- 238000010008 shearing Methods 0.000 abstract description 43
- 230000000694 effects Effects 0.000 abstract description 13
- 210000004027 cell Anatomy 0.000 description 43
- 238000005452 bending Methods 0.000 description 25
- 239000000725 suspension Substances 0.000 description 17
- 230000001464 adherent effect Effects 0.000 description 14
- 239000012531 culture fluid Substances 0.000 description 12
- 238000010586 diagram Methods 0.000 description 12
- 238000000034 method Methods 0.000 description 9
- 230000002776 aggregation Effects 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 238000004088 simulation Methods 0.000 description 8
- 210000004102 animal cell Anatomy 0.000 description 7
- 230000035755 proliferation Effects 0.000 description 5
- 238000005054 agglomeration Methods 0.000 description 4
- 238000004220 aggregation Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 235000006481 Colocasia esculenta Nutrition 0.000 description 2
- 241001149904 Elliptio crassidens Species 0.000 description 2
- 235000013289 Xanthosoma Nutrition 0.000 description 2
- 238000000418 atomic force spectrum Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000004114 suspension culture Methods 0.000 description 2
- 238000013019 agitation Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003822 cell turnover Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000003631 expected effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
- B01F27/05—Stirrers
- B01F27/11—Stirrers characterised by the configuration of the stirrers
- B01F27/112—Stirrers characterised by the configuration of the stirrers with arms, paddles, vanes or blades
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- Chemical & Material Sciences (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Organic Chemistry (AREA)
- Zoology (AREA)
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- General Engineering & Computer Science (AREA)
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- Genetics & Genomics (AREA)
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a stirring wheel and a bioreactor, wherein the stirring wheel comprises a wheel hub, a plurality of blades and a blade group, the wheel hub is in a hollow cylindrical shape, openings are formed at the opposite ends of the wheel hub, and at least two openings are formed in the circumferential direction of the wheel hub at intervals; the blades are distributed on the peripheral wall of the hub at intervals; the blade group is installed inside the hub, and the blade group is used for enabling fluid inside the hub to flow in the axial direction of the hub. According to the technical scheme, the culture solution in the bioreactor is stirred by adopting the stirring wheel with the blades circumferentially arranged on the peripheral wall of the hub at intervals, under the condition of reducing the shearing force of the stirring wheel on cells, the uniformity of a flow field and the turning quantity of the cells are improved, the hub is provided with the holes, the blade group is arranged in the hub, so that the culture solution in the hub can be uniformly mixed with the culture solution outside the hub, and the bioreactor with the stirring wheel in a larger working volume still has a better stirring effect.
Description
Technical Field
The invention relates to the technical field of stirring wheels, in particular to a stirring wheel and a bioreactor.
Background
Generally, the domesticated adherent animal cells or suspension cells have a process of growing continuously in an agglomeration manner without destroying the agglomeration during a long-period culture. In the whole culture period, the stirring rotation speed needs to be adjusted in real time according to the cell aggregation characteristic, and meanwhile, in the adjustment process, the requirement of extremely low shearing force needs to be met in the whole process, so that the activity of cells is not influenced due to overlarge shearing force, and the cell aggregation and the final proliferation effect are damaged.
At present, most of lifting stirring paddles are used for culturing carrier-free suspension cells or cell balls, and the stirring paddles are composed of a plurality of smaller elephant ear paddles, so that a better flow field can be provided only under a relatively large stirring rotation speed, and the requirement of cell suspension culture is met. However, the lifting type stirring paddle can only be used for bioreactors with smaller working volumes (culture solution volumes), and cannot meet the requirements of bioreactors with larger working volumes on flow field uniformity and cell turnover. If the rotation speed of the stirring paddle is increased more, the shearing force is exponentially amplified, the requirement of the suspension cell ball culture on extremely low shearing force cannot be met, and then the cell culture proliferation effect is possibly poor, so that the culture expectation cannot be reached.
Disclosure of Invention
The invention mainly aims to provide a stirring wheel which aims to improve the uniformity of a flow field in a bioreactor.
To achieve the above object, the present invention provides a bioreactor comprising:
the hub is arranged in a hollow cylindrical shape, openings are formed in the two opposite ends of the hub, and at least two openings are formed in the circumferential direction of the hub at intervals;
the blades are respectively connected to the hub and are distributed on the peripheral wall of the hub at intervals;
and the blade group is arranged in the hub and is used for enabling fluid in the hub to flow in the axial direction of the hub.
In an embodiment, a plurality of the blades extend in the axial direction of the hub, the blades comprise straight plate sections and bending sections which are connected, the bending sections are located at the ends of the blades, and the bending sections are bent in the same direction along the circumferential direction of the hub.
In one embodiment, the openings are located between any two adjacent vanes.
In one embodiment, the number of the openings is two, and the two openings are symmetrically distributed on the hub.
In one embodiment, the number of the holes is two, and the two holes are distributed on the hub in a staggered manner.
In an embodiment, an included angle formed by the two openings and a connecting line of the hub axle is greater than or equal to 90 °.
In an embodiment, the aperture extends in an axial direction of the hub.
In one embodiment, the length of the opening is set to L, and the axial length of the hub is set to H, L/H satisfies the range of 1/2 to 1.
In an embodiment, the opening comprises a plurality of liquid passing ports, and the liquid passing ports are distributed at intervals in the axial direction of the hub.
In an embodiment, the length of the blade coincides with the hub axial length.
In one embodiment, the number of blades is set to 4 to 20.
In one embodiment, the number of blades is set to 8 to 16.
In an 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 an axial direction of the hub.
In an embodiment, the upper blade group includes two first blades, the first blades are connected to the inner peripheral wall of the hub, and the two first blades are distributed at intervals in the circumferential direction of the inner peripheral wall of the hub; the lower blade group comprises two second blades, the second blades are connected to the inner peripheral wall of the hub, and the two second blades are distributed at intervals in the circumferential direction of the inner peripheral wall of the hub.
The invention also provides a bioreactor, which comprises a hub, a plurality of blades and a blade group, wherein the hub is arranged in a hollow cylinder shape, openings are formed at the opposite ends of the hub, and at least two openings are formed at intervals in the circumferential direction of the hub; the blades are respectively connected to the hub, and are distributed on the peripheral wall of the hub at intervals; the blade group is mounted inside the hub, and the blade group is used for enabling fluid inside the hub to flow in the axial direction of the hub.
According to the technical scheme, the culture solution in the bioreactor is stirred by adopting the stirring wheel with the blades circumferentially arranged on the peripheral wall of the hub at intervals, so that the uniformity of a flow field and the turning quantity of cells can be improved under the condition of reducing the shearing force of the stirring wheel on the cells, the hub is provided with the holes, the blade group is arranged in the hub, so that the culture solution in the hub can be uniformly mixed with the culture solution outside the hub, and the bioreactor with the stirring wheel in a larger working volume still has a better stirring effect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to the structures shown in these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a velocity cloud of a bioreactor (bioreactor working volume 500 ml) stirred with an ear paddle;
FIG. 2 is a velocity cloud of a bioreactor stirred with an ear paddle (bioreactor working volume 5L) in the background;
FIG. 3 is a velocity cloud of a bioreactor (bioreactor working volume 10L) with a paddle stirring in the background;
FIG. 4 is a schematic view of the structure of the stirring wheel in one embodiment;
FIG. 5 is a schematic view of the structure of FIG. 4 at another view angle;
FIG. 6 is a schematic view of the structure of the stirring wheel in a cross-sectional view;
FIG. 7 is a schematic view of the structure of the bioreactor at a cross-sectional view;
FIG. 8 is a schematic view of the agitator wheel in another cross-sectional view;
FIG. 9 is a graph of shear force distribution for a hub without openings;
FIG. 10 is a graph of shear force profile for a hub with an opening;
FIG. 11 is a graph of shear force profile for a hub having two openings;
FIG. 12 is a graph of shear force distribution for a hub with four openings;
FIG. 13 is a velocity cloud of a bioreactor with a working volume of 5L in elevation;
FIG. 14 is a velocity cloud of a bioreactor with a working volume of 10L in elevation;
FIG. 15 is a velocity cloud of a bioreactor with a working volume of 5L in plan view;
FIG. 16 is a velocity cloud of a bioreactor with a working volume of 10L in plan view;
FIG. 17 is a velocity cloud of a bioreactor with a working volume of 5L in bottom view;
FIG. 18 is a velocity cloud of a bioreactor with a working volume of 10L in bottom view;
FIG. 19 is a velocity cloud of the bioreactor in front view with two openings symmetrically distributed;
FIG. 20 is a velocity cloud of the bioreactor in plan view with two openings symmetrically distributed;
FIG. 21 is a velocity cloud of the bioreactor in bottom view with two openings symmetrically distributed;
FIG. 22 is a velocity cloud of the bioreactor in elevation with two openings in offset distribution;
FIG. 23 is a velocity cloud of the bioreactor in plan view with two openings offset;
FIG. 24 is a velocity cloud of the bioreactor in bottom view with two openings in offset distribution;
FIG. 25 is a velocity cloud of the bioreactor in elevation view at a blade count of 4;
FIG. 26 is a velocity cloud of the bioreactor in elevation view at a blade count of 6;
FIG. 27 is a velocity cloud of the bioreactor in elevation at 8 blades;
FIG. 28 is a velocity cloud of the bioreactor in elevation view at a blade count of 12;
FIG. 29 is a velocity cloud of the bioreactor in elevation view with a blade count of 16;
FIG. 30 is a velocity cloud of the bioreactor at 20 blades in elevation;
FIG. 31 is a velocity cloud of the bioreactor in plan view with a number of blades of 4;
FIG. 32 is a velocity cloud of the bioreactor in plan view with a number of blades of 6;
FIG. 33 is a velocity cloud of the bioreactor in plan view with a blade count of 8;
FIG. 34 is a velocity cloud of the bioreactor in plan view with a number of blades of 12;
FIG. 35 is a velocity cloud of the bioreactor in plan view with a number of blades of 16;
FIG. 36 is a velocity cloud of the bioreactor in plan view with a blade count of 20.
Reference numerals illustrate:
the achievement of the objects, functional features and advantages of the present invention will be further described with reference to the accompanying drawings, in conjunction with the embodiments.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that, if directional indications (such as up, down, left, right, front, and rear … …) are included in the embodiments of the present invention, the directional indications are merely used to explain the relative positional relationship, movement conditions, etc. between the components in a specific posture (as shown in the drawings), and if the specific posture is changed, the directional indications are correspondingly changed.
In addition, if there is a description of "first", "second", etc. in the embodiments of the present invention, the description of "first", "second", etc. is for descriptive purposes only and is not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In addition, if the meaning of "and/or" is presented throughout this document, it is intended to include three schemes in parallel, taking "a and/or B" as an example, including a scheme, or B scheme, or a scheme where a and B meet simultaneously. In addition, the technical solutions of the embodiments may be combined with each other, but it is necessary to base that the technical solutions can be realized by those skilled in the art, and when the technical solutions are contradictory or cannot be realized, the combination of the technical solutions should be considered to be absent and not within the scope of protection claimed in the present invention.
The domesticated adherent animal cells or suspension cells have the technological process of continuous agglomeration and growth without destroying agglomeration in the long-period culture process. In the whole culture period, the stirring rotation speed needs to be adjusted in real time according to the cell aggregation characteristic, and meanwhile, in the adjustment process, the requirement of extremely low shearing force needs to be met in the whole process, so that the activity of cells is not influenced due to overlarge shearing force, and the cell aggregation and the final proliferation effect are damaged.
At present, a traditional lifting type stirring paddle is used for culturing carrier-free suspension cells or cell balls, the stirring paddle is composed of a plurality of smaller elephant ear paddles, and a better flow field can be provided only under a relatively large stirring rotation speed, so that the requirement of cell suspension culture is met. The higher rotation speed can cause the exponential amplification of the shearing force, and the extremely low shearing force requirement of the suspension cell ball culture can not be met, so that the cell culture proliferation effect can be poor, and the culture expectation can not be reached.
Referring to fig. 1 to 3, fig. 1 to 3 are flow field velocity simulation diagrams of a bioreactor having a lifting paddle. The flow rate and the color correspond to the color areas in the simulated image and the right bioreactor, and generally, the area close to the stirring paddle is lighter in color, representing the higher flow rate, the area far away from the stirring paddle is darker, and representing the lower flow rate.
FIG. 1 is a flow field velocity simulation diagram of a bioreactor with a working volume of 500ml, and it can be seen that the flow velocity around the stirring paddle is higher, the flow field velocity is uniformly distributed, and the flow field uniformity is better. Fig. 2 and 3 are flow field velocity simulation diagrams of the bioreactor with working volumes of 5L and 10L, respectively, and it is obvious that the flow velocity around the stirring paddle is lower, the flow field velocity distribution is uneven, and the mixing degree of the flow field is poor compared with fig. 1. It follows that such lifting paddles are not suitable for bioreactors of large working volume.
In view of this, the present invention proposes a stirring wheel 10.
In the embodiment of the present invention, as shown in fig. 4 to 8, the stirring wheel 10 includes a hub 100, a plurality of blades 200 and a blade set 300, wherein the hub 100 is in a hollow cylindrical shape, two opposite ends of the hub 100 are provided with openings 110, and at least two openings 120 are arranged on the hub 100 at intervals in the circumferential direction; the plurality of blades 200 are respectively connected to the hub 100, and the plurality of blades 200 are distributed on the outer peripheral wall of the hub 100 at intervals; the blade group 300 is mounted inside the hub 100, and the blade group 300 is used to flow the fluid inside the hub 100 in the axial direction of the hub 100.
Specifically, the agitation wheel 10 in this embodiment is used in a bioreactor to agitate the culture solution and cells in the bioreactor so that the adherent animal cells or the suspension cells can obtain the desired gas and culture solution. The stirring wheel 10 of the invention generally sets the rotating speed between 5rpm and 50rpm when culturing adherent animal cells or suspension cells, can ensure that various culture solutions in the bioreactor are fully mixed at the rotating speed, and can meet the basic turnover amount required by the cells, and the average shearing force is between 0.03Pa and 0.2Pa when the stirring wheel 10 is in the rotating speed range so as to meet the requirement of the adherent animal cells or suspension cells on low shearing force.
The hub 100 is formed in a hollow cylindrical shape, 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, the plurality of blades 200 extending in the axial direction of the hub 100, and in consideration of the fact that the blades 200 on the hub 100 are required to have sufficient stirring force, the number of the blades 200 cannot be set too small, and the number of the blades 200 is at least set to be greater than or equal to four, and thus the radial length of the hub 100 cannot be set too small to satisfy the requirement of having sufficient positions to mount the plurality of blades 200. The shape of the blade 200 may be a straight plate, an arc plate, or a special shape, which is not particularly limited herein.
In order to make the culture solution inside the hub 100 fully mix and turn, the stirring wheel 10 further includes a blade set 300, where the blade set 300 is disposed inside the hub 100, and the blade set 300 can make the culture solution inside the hub 100 flow in the axial direction of the hub 100, where, in order to make the flow effect of the culture solution inside the hub 100 better, the blade set 300 may be provided with a plurality of blades, where the plurality of blades may be circumferentially disposed inside the hub 100, and the plurality of blades may also be axially disposed both along the inner circumference of the hub 100 and along the inner circumference of the hub 100, where the arrangement of the blades inside the blade set 300 is not specifically limited.
Further, the flow of the culture solution inside the hub 100 is balanced with the flow of the culture solution outside the hub 100, so that the culture solution in the two parts is more uniformly mixed, and at least two openings 120 are formed on the inner wall of the hub 100. It should be noted that, referring to fig. 9 to 12, the shear force distribution diagram of the bioreactor in fig. 9 to 12 is that the original shear force changes from low to high (i.e. from bottom to top in the drawing) from blue to yellow and then from red, after the gray scale treatment is performed on the simulated image, the shear force changes from low to high, and changes from dark to light and then from dark, wherein the shear force and the color correspond to the color areas in the simulated image and the bioreactor on the right, and generally, the area near the stirring paddle has lighter color, represents higher shear force, the area far from the stirring paddle is deeper, and represents lower shear force.
Wherein, fig. 9 is a comparison scheme of not providing an opening 120 on the hub 100, fig. 10 is a comparison scheme of providing one opening 120 on the hub 100, and fig. 11 is a scheme of providing two openings 120 on the hub 100 in this embodiment, fig. 12 is a scheme of providing two openings 120 on the hub 100, it is obvious in fig. 9 that the inner area of the hub 100 is very dark, which represents the very low shearing force inside the hub 100, and the inner area of the hub 100 is partially dark, which represents the uneven distribution of shearing force inside the hub 100, so that the two comparison schemes of fig. 9 and 10 may cause poor turning of adherent cells or suspension cells inside the hub 100. Whereas the color of the interior of hub 100 in fig. 11 is much brighter than fig. 9 and 10, indicating higher shear forces within hub 100. In view of this, at least two openings 120 should be formed in the hub 100.
Referring to fig. 13 to 18, where fig. 13, 15 and 17 are flow rate cloud diagrams of the present embodiment under a working volume of 5L, fig. 14, 16 and 18 are flow rate cloud diagrams of the present embodiment under a working volume of 10L, it should be noted that the left side of the simulation diagram is a numerical change of flow rate, the original flow rate is changed from low to high (i.e. from bottom to top in the diagram) from blue to yellow to red, after the simulation diagram is subjected to gray scale treatment, the flow rate is changed from low to high to dark to light to dark, wherein the flow rate color corresponds to the color region in the bioreactor on the right side, in general, the color of the region close to the stirring wheel 10 and the blade 200 is lighter, which represents the flow rate is higher, the region far from the stirring wheel 10 is darker, which represents the flow rate is lower, and the flow rate inside the stirring wheel 10 is smaller than the flow rate outside the stirring wheel 10. If a dark region appears in the middle of a light region around the agitator wheel 10, this represents a light region where the flow rate is higher than the periphery. The description applies equally to the flow rate cloud in the following examples, and is therefore only described once here.
It can be seen that comparing fig. 13 and 14 with fig. 2 and 3, the lifting type stirring paddle of the present embodiment has good flow field uniformity under a large working volume, so that the stirring wheel of the present embodiment is better selected under a large working volume.
Further, in combination with the simulation data provided in the table below, the rotation speed of the lifting type stirring paddle was 60rpm, the stirring paddle was divided into an upper layer stirring paddle and a lower layer stirring paddle, the average shear rate around the upper layer stirring paddle was 12.396, the average shear rate around the lower layer stirring paddle was 12.388, and there was a point around the upper layer stirring paddle and around the lower layer stirring paddle having the maximum shear force, where the maximum shear rate was 224.7, and the average shear rate of the other areas in the tank body except the upper layer stirring paddle and the lower layer stirring paddle was 3.137.
The flow field velocity simulation diagram of the bioreactor in fig. 2 is that the working volume is 5L, and it can be seen that the flow field velocity of the stirring paddle bioreactor is higher and the flow field uniformity is better. In combination with the simulation data provided in the table below, the rotational speed of the lifting paddles was 60rpm, the paddles were divided into upper and lower paddles, the average shear rate around the upper paddle was 12.289, the average shear rate around the lower paddle was 14.210, and there was a point around the upper and lower paddles with the maximum shear force, where there was 631.4, and the average shear rate of the other areas in the tank except the upper and lower paddles was 2.128. In the table, the average shear force around the plurality of blades of the stirring wheel of this example was 4.098, the average shear force in the other region was 2.781, and the shear force at the maximum point was 99.41. It can be derived that the average shearing force of the stirring wheel 10 around the plurality of blades 200 of the stirring wheel 10 is smaller than the average shearing force of the lifting type stirring paddle around the stirring paddle, and the shearing force of the maximum point of the stirring wheel 10 is much smaller than the shearing force of the maximum point of the lifting type stirring paddle, and therefore, the low shearing force of the stirring wheel 10 of the present embodiment is more suitable for the culture environment required by the adherent cells or the suspension cells.
According to the technical scheme, the culture solution in the bioreactor is stirred by adopting the stirring wheel 10 with the blades 200 arranged on the peripheral wall of the hub 100 at intervals, so that the uniformity of a flow field and the turning quantity of cells can be improved under the condition of reducing the shearing force of the stirring wheel 10 on the cells, the hub 100 is provided with the open holes 120, the blade group 300 is arranged in the hub 100, the culture solution in the hub 100 can be uniformly mixed with the culture solution outside the hub 100, and the bioreactor with the stirring wheel 10 in the large working volume still has a good stirring effect.
In an embodiment, referring to fig. 5 to 7, a plurality of the blades 200 extend in an axial direction of the hub 100, the blades 200 include a straight plate section 210 and a bent section 220 connected to each other, the bent section 220 is located at an end of the blades 200, and the bent section 220 is bent in a circumferential direction of the hub 100.
It is considered that if the entire blade 200 is formed in a straight plate shape, the cells are directly stirred, and the adherent cells cannot be cultured well. Further, the blade 200 includes a straight plate section 210 and a bending section 220 connected, wherein the bending section 220 is located at an end of the blade 200, and the bending section 220 is bent along a circumferential direction of the hub 100. The provision of the straight plate section 210 can expand the stirring range of the blade 200 and sufficiently stir the culture solution in the bioreactor. In the process of rotating the stirring wheel 10, the bending section 220 is easy to set so that the stirring wheel 10 drives a part of adherent cells or suspension cells to rotate together, and the culture cells can be fully and uniformly mixed with the culture solution to absorb nutrient substances and required gases in the culture solution. And the bending section 220 is bent along the circumferential direction of the hub 100, so that the bending direction of the bending section 220 is consistent with the rotation track of the stirring wheel 10, and the cultured cells are easier to drive to rotate together.
Further, referring to fig. 4 to 8, the opening 120 is located between any two adjacent blades 200.
It should be noted that, in order to facilitate opening the openings 120, the openings 120 are opened between any two adjacent blades 200, and corresponding to each other, other openings 120 are opened between any two adjacent blades 200, where two or more openings 120 may be located between the same two adjacent blades 200 or between different two adjacent blades 200, and in consideration of that the openings 120 are circumferentially spaced, it is a preferred embodiment to open the openings 120 between different two adjacent blades 200.
In an embodiment, please refer to fig. 5, 19, 20 and 21, wherein fig. 19 to 21 are flow field velocity profiles when the openings 120 are symmetrically arranged, wherein fig. 19 is a velocity cloud for a side cross-sectional view, fig. 20 is a velocity cloud for an upper side view, and fig. 21 is a velocity cloud for a lower side view. The number of the openings 120 is two, and the two openings 120 are symmetrically distributed on the hub 100.
Specifically, when two openings 120 are formed, the culture fluid flowing into the hub 100 from one opening 120 can flow out from the other opening 120, that is, the culture fluid in the hub 100 can flow not only in the axial direction but also in the radial direction of the hub 100, which enhances the uniformity of the culture fluid in the flow channel in the hub 100 and the culture fluid outside the hub 100, and also makes the flow field velocity distribution in the hub 100 more uniform, and it can be seen from fig. 19 to 21 that the flow velocity distribution in the hub 100 and the flow velocity distribution outside the hub 100 are more uniform.
In an embodiment, please refer to fig. 8, 22, 23 and 24, wherein fig. 23 to 24 are flow field velocity profiles when the pairs of apertures 120 are arranged, wherein fig. 22 is a velocity cloud for a side cross-sectional view, fig. 23 is a velocity cloud for an upper side view, and fig. 24 is a velocity cloud for a lower side view. The number of the openings 120 is two, and the two openings 120 are distributed on the hub 100 in a staggered manner.
Specifically, when two openings 120 are formed, the culture fluid flowing into the hub 100 from one opening 120 can flow out from the other opening 120, that is, the culture fluid in the hub 100 can flow not only in the axial direction but also in the radial direction of the hub 100, which enhances the uniformity of the culture fluid in the flow channel in the hub 100 and the culture fluid outside the hub 100, and also makes the flow field velocity distribution in the hub 100 more uniform, and it can be seen from fig. 19 to 21 that the flow velocity distribution in the hub 100 and the flow velocity distribution outside the hub 100 are more uniform.
In one embodiment, referring to fig. 8, the included angle formed by the two openings 120 and the axis of the hub 100 is greater than or equal to 90 °.
Considering that, when the included angle formed by the connection line between the two openings 120 and the axle center of the hub 100 is smaller than 90 °, the culture solution is not easy to flow out of one opening 120 through the other opening 120 after flowing into the hub 100, which may cause the slowing of the flow velocity in the hub 100 or uneven flow velocity, and further cause poor or uneven turning amount, so that the culture of adherent cells or suspension cells is difficult to achieve the expected effect.
In one embodiment, referring to fig. 5-6, the opening 120 extends in the axial direction of the hub 100.
In order to further improve the culture fluid communication between the outside of the hub 100 and the 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, so that the culture fluid interaction effect between the opening 120 and the hub 100 is improved.
Further, referring to fig. 5, the length of the opening 120 is set to L, and the axial length of the hub 100 is set to H, L/H satisfies the range 1/2 to 1.
Specifically, the length of the openings 120 is such that the ratio of the axial length of the hub 100 to the axial length of the hub 100 is at least 1/2
1, i.e., the length of the opening 120 may be greater than or equal to 1/2 of the axial length of the hub 100, and the length of the opening 120 may be less than the axial length of the hub 100, in which range the culture fluid outside the hub 100 and inside the hub 100 may be allowed to flow through and interact, so that nutrients of each component in the culture fluid may flow through each other.
In an embodiment, referring to fig. 9, the opening 120 includes a plurality of liquid passing ports, and the liquid passing ports are spaced apart in an axial direction of the hub 100.
It should be noted that, the present invention is not limited to the above embodiment, the plurality of openings 120 are circumferentially spaced apart, and further, the openings 120 may include a plurality of liquid passing openings, and the plurality of liquid passing openings are circumferentially spaced apart and arranged in the axial direction of the hub 100, and the plurality of pipes are inserted in consideration of different depth positions in the bioreactor, so that the main components of the nutrient substances entering each liquid passing opening can be controlled, so that the inflow components of different liquid passing openings are different.
In an embodiment, referring to fig. 4, the bending direction of the bending sections 220 is consistent with the rotation direction of the hub 100.
In the above embodiment, the bending sections 220 are limited to bend circumferentially, in order to further limit the bending directions of the bending sections 220 in the circumferential bending direction, the bending directions of the bending sections 220 are consistent with the rotation direction of the hub 100, when the hub 100 rotates clockwise, the bending sections 220 bend clockwise, when the multi-hub 100 rotates anticlockwise, the bending sections 220 rotate anticlockwise, so that the bending directions of the bending sections 220 are consistent, and the bending directions of the bending sections 220 are the same as the rotation directions, so that the stirring wheel 10 is easier to drive the adherent cells or suspension cells in the bending sections to rotate together during stirring.
In one embodiment, referring to fig. 5 and 6, the blade 200 has a length that corresponds to the axial length of the hub 100.
In order to make the blade 200 have a larger rotation range, the length of the blade 200 is enlarged as much as possible, but if the length of the blade 200 exceeds the axial length of the hub 100, the portion of the blade 200 exceeding the hub 100 is not connected to the hub 100, which may cause unstable rotation of the stirring wheel 10, so that the length of the blade 200 is consistent with the axial length of the hub 100, the length of the blade 200 may be the same as the axial length of the hub 100, the length of the blade 200 may be slightly greater than the axial length of the hub 100, or the length of the blade 200 may be slightly less than the axial length of the hub 100.
In one embodiment, referring to fig. 25 to 36, the number of the blades 200 is set to 4 to 20.
Wherein fig. 25 to 30 are velocity cloud diagrams of the bioreactor in which the number of bioreactor blades 200 is gradually increased in a front view, and fig. 31 to 36 are velocity cloud diagrams of the bioreactor in which the number of bioreactor blades 200 is gradually increased in a top view.
It is apparent from the front view and the top view that when the number of the blades 200 is 4 and 6, the flow velocity distribution around the blades 200 is significantly less uniform than when the number of the blades 200 is 8 to 20. And when the number of the blades 200 is 12, the uniformity of the flow field as a whole is the best.
With continued reference to the following table, shear force data in the bioreactor for the number of blades 200 of 4, 6, 8, 12, 16, 20. When the number of blades 200 is 4, the average shearing force around the blades 200 is 3.998, the average shearing force in other areas is 1.534, and the maximum shearing force is 40.940. When the number of the blades 200 is 6, the average shearing force around the blades 200 is 4.276, the average shearing force in other areas is 1.634, and the maximum shearing force is 22.938. When the number of blades 200 is 8, the average shearing force around the blades 200 is 4.416, the average shearing force in other areas is 1.734, and the maximum shearing force is 33.042. When the number of blades 200 is 12, the average shearing force around the blades 200 is 4.622, the average shearing force in other areas is 1.707, and the maximum shearing force is 29.678. When the number of blades 200 is 16, the average shearing force around the blades 200 is 4.721, the average shearing force in other areas is 1.611, and the maximum shearing force is 45.478. When the number of the blades 200 is 20, the average shearing force around the blades 200 is 4.510, the average shearing force in other areas is 1.635, and the maximum shearing force is 51.610.
From the above table and data, when the number of the blades 200 is 4 or 6, the average shearing force around the blades 200 is too small to achieve a good effect of turning the cells, and when the number of the blades 200 is 16 or 20, the maximum shearing force is too large, which may inhibit the growth and proliferation of adherent animal cells or suspension cells. When the number of the blades 200 is 12, the shearing force around the blades 200 is suitable, and the maximum shearing force is also small, so that the number of the blades 200 is set to 12, which is the best embodiment of the present embodiment.
Referring to the influence of the number of blades 200 on the flow field velocity distribution in the bioreactor and the effect of the number of blades 200 on the shearing force, it can be obtained that the preferred embodiment of the present embodiment is when the number of blades 200 is set to 12, and of course, the basic requirement of culturing adherent cells or suspension cells can be satisfied when the number of blades 200 is set to other numbers in the range of 4 to 20.
In one embodiment, referring to fig. 25 to 36, the number of the blades 200 is set to 8 to 16.
The number of blades 200 is set to 8 to 16.
Further, referring to the above analysis and tables, culture of adherent animal cells and suspension cells requires as low a shear force as possible and a good uniformity of flow rate, and thus, setting the number of blades 200 to 8 to 16 is a more preferable example of the above embodiment.
In an embodiment, referring to fig. 6, the blade set 300 includes an upper blade set 310 and a lower blade set 320, and the upper blade set 310 and the lower blade set 320 are spaced apart in the axial direction of the hub 100.
In order to further increase the flow rate of the culture medium and the amount of the cells in the blade group 300, the blade group 300 is divided into an upper blade group 310 and a lower blade group 320, and the culture medium in the hub 100 is agitated by the upper blade group 310 and the lower blade group 320, respectively, so that the amount of the cells in the hub 100 is increased, considering that the hub 100 has a certain length in the axial direction thereof. The upper layer blade set 310 and the lower layer blade set 320 are arranged at intervals in the axial direction of the hub 100, and the upper layer blade set 310 and the lower layer blade set 320 are close to the two openings 110 of the hub 100 as much as possible, so that the upper layer blade set 310 and the lower layer blade set 320 stir and turn the culture solution in the hub 100 as much as possible.
Further, referring to fig. 6, 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, where 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.
Specifically, in the present embodiment, the upper blade group 310 includes two first blades 311, which is not limited to the present embodiment, and of course, the upper blade group 310 may also include three first blades 311, and the upper blade group 310 may also include four first blades 311 or even more first blades 311. Wherein, two first paddles 311 are spaced apart in the circumferential direction of the inner circumferential wall of the hub 100 to improve the stirring ability of the upper paddle set 310. Correspondingly, in the present embodiment, the lower blade set 320 includes two second blades 321, which is not limited to the present embodiment, and of course, the lower blade set 320 may also include three second blades 321, and the lower blade set 320 may also include four second blades 321 or even more second blades 321. Wherein, two second paddles 321 are spaced apart in the circumferential direction of the inner circumferential wall of the hub 100 to improve the stirring ability of the lower paddle set 320.
In addition, the upper blade group 310 and the lower blade group 320 are also connected to the inner peripheral wall of the hub 100, and thus the upper blade group 310 and the plurality of blades 200 outside the hub 100 can rotate together, which can be achieved by only one driving device, and the driving method of the stirring wheel 10 is simplified.
The invention also provides a bioreactor, which comprises a stirring wheel 10, wherein the specific structure of the stirring wheel 10 refers to the embodiment, and as the bioreactor adopts all the technical schemes of all the embodiments, the bioreactor at least has all the beneficial effects brought by the technical schemes of the embodiments, and the description is omitted herein.
The foregoing description is only of the optional embodiments of the present invention, and is not intended to limit the scope of the invention, and all the equivalent structural changes made by the description of the present invention and the accompanying drawings or the direct/indirect application in other related technical fields are included in the scope of the invention.
Claims (15)
1. A stirring wheel (10) for a bioreactor, characterized by comprising:
the hub (100) is arranged in a hollow cylindrical shape, openings (110) are formed at two opposite ends of the hub (100), and at least two holes (120) are formed in the hub (100) at intervals in the circumferential direction of the hub;
a plurality of blades (200), the plurality of blades (200) being respectively connected to the hub (100), the plurality of blades (200) being spaced apart on an outer peripheral wall of the hub (100);
-a blade set (300), the blade set (300) being mounted inside the hub (100), the blade set (300) being for flowing a fluid inside the hub (100) in an axial direction of the hub (100).
2. The stirring wheel (10) according to claim 1, characterized in that a plurality of said blades (200) extend in the axial direction of said hub (100), said blades (200) comprising connected straight plate sections (210) and bent sections (220), said bent sections (220) being located at the ends of said blades (200), a plurality of said bent sections (220) being bent in the same direction along the circumference of said hub (100).
3. The stirring wheel (10) according to claim 2, characterized in that the aperture (120) is located between any two adjacent blades (200).
4. A mixer wheel (10) according to claim 3, wherein the number of openings (120) is two, two openings (120) being symmetrically distributed on the hub (100).
5. A mixer wheel (10) according to claim 3, wherein the number of openings (120) is two, and wherein two openings (120) are offset in the hub (100).
6. The stirring wheel (10) according to claim 5, characterized in that the two openings (120) form an angle with the axis of the hub (100) of greater than or equal to 90 °.
7. A stirring wheel (10) according to claim 3, characterized in that the aperture (120) extends in the axial direction of the hub (100).
8. A stirring wheel (10) according to claim 3, characterized in that the length of the aperture (120) is set to L and the axial length of the hub (100) is set to H, L/H satisfying the range 1/2 to 1.
9. A mixer wheel (10) according to claim 3, wherein the aperture (120) comprises a plurality of through openings, which are spaced apart in the axial direction of the hub (100).
10. The stirring wheel (10) according to claim 2, characterized in that the length of the blades (200) coincides with the axial length of the hub (100).
11. The stirring wheel (10) according to claim 2, characterized in that the number of blades (200) is set to 4 to 20.
12. The stirring wheel (10) according to claim 11, characterized in that the number of blades (200) is set to 8 to 16.
13. The stirring wheel (10) according to claim 2, wherein the blade set (300) comprises an upper blade set (310) and a lower blade set (320), the upper blade set (310) and the lower blade set (320) being spaced apart in an axial direction of the hub (100).
14. The stirring wheel (10) according to claim 13, wherein the upper blade group (310) comprises two first blades (311), the first blades (311) being connected to the inner circumferential wall of the hub (100), the two first blades (311) being spaced apart in the circumferential direction of the inner circumferential wall of the hub (100); the lower blade group (320) comprises two second blades (321), the second blades (321) are connected to the inner peripheral wall of the hub (100), and the two second blades (321) are distributed at intervals in the circumferential direction of the inner peripheral wall of the hub (100).
15. Bioreactor, characterized in that it comprises a stirring wheel (10) according to any one of claims 1 to 14.
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| CN117431144B (en) | 2024-06-21 |
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