CN213447165U - Integrated rotary stirring filter for cell culture - Google Patents

Integrated rotary stirring filter for cell culture Download PDF

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Publication number
CN213447165U
CN213447165U CN202022278644.0U CN202022278644U CN213447165U CN 213447165 U CN213447165 U CN 213447165U CN 202022278644 U CN202022278644 U CN 202022278644U CN 213447165 U CN213447165 U CN 213447165U
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fixed seat
stirring shaft
stirring
air outlet
frame
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CN202022278644.0U
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Chinese (zh)
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王宪其
王洁筠
李启照
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NANJING JINRI LIGHT INDUSTRY TECHNOLOGY DEVELOPMENT CO LTD
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NANJING JINRI LIGHT INDUSTRY TECHNOLOGY DEVELOPMENT CO LTD
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Abstract

The utility model provides an integrated rotary stirring filter for cell culture, which has low shearing effect, prevents cell damage, promotes cell growth, has less foam and good mixing performance, comprises a stirring shaft and a stirring paddle, and also comprises a cage-shaped gas exchanger consisting of a frame-type cylinder, an upper flange and a lower flange which are connected with both ends of the frame-type cylinder, and a screen fixed at the periphery of the frame-type cylinder; the mesh size of the screen is 10um-75 um; the inner bottom of the cage-type gas exchanger is provided with an air outlet pipe, air outlet holes are uniformly distributed on the air outlet pipe, and the air outlet pipe is communicated with a communicating pipe which extends upwards through the upper flange plate; the stirring paddle comprises a fixed seat and a circular tube which is connected to the periphery of the fixed seat and is used as a paddle; the upper end of the fixed seat is connected with a stirring shaft, and the lower end of the fixed seat is connected with an upper flange plate; an air inlet channel is arranged in the stirring shaft along the axis of the stirring shaft, and the air inlet channel is communicated with the communicating pipe through a transition channel passing through the fixed seat.

Description

Integrated rotary stirring filter for cell culture
Technical Field
The utility model relates to a be applied to bioreactor and cell culture's rotatory stirring filter.
Background
Filters used in the field of sterile culture such as magnetic suspension process perfusion equipment are generally mounted on another shaft, extend into tanks (biological reaction tanks, cell culture tanks and the like), and the shaft is fixed. Thus, the dissolved oxygen in the tank cannot be fully and uniformly dissolved. Once the stirring function is started, a large amount of bubbles are generated, the cells are crushed by the bubbles, and the cells are lost to a certain extent in the cell culture process.
The animal cells are not protected by cell walls and are therefore very sensitive to shearing, and are easily damaged by direct mechanical agitation, which is obviously unsuitable for the culture of animal cells in conventional stirred reactors for microorganisms.
Disclosure of Invention
The utility model provides a have the low shear effect, prevent cell damage, can promote cell growth, the foam is few, and the rotatory stirring filter of integral type for cell culture that mixing performance is good is applicable to the cell and realizes high density culture in extensive bioreactor.
In order to achieve the above purpose, the integrated rotary stirring filter for cell culture of the utility model comprises a stirring shaft, a stirring paddle, a cage-type gas exchanger consisting of a frame-type cylinder, an upper flange and a lower flange connected with both ends of the frame-type cylinder, and a screen fixed on the periphery of the frame-type cylinder; the mesh size of the screen is 10um-75 um; the inner bottom of the cage-type gas exchanger is provided with an air outlet pipe, air outlet holes are uniformly distributed on the air outlet pipe, and the air outlet pipe is communicated with a communicating pipe which extends upwards through the upper flange plate; the stirring paddle comprises a fixed seat and a circular tube which is connected to the periphery of the fixed seat and is used as a paddle; the upper end of the fixed seat is connected with a stirring shaft, and the lower end of the fixed seat is connected with an upper flange plate; an air inlet channel is arranged in the stirring shaft along the axis of the stirring shaft, and the air inlet channel is communicated with the communicating pipe through a transition channel passing through the fixed seat.
As a further improvement of the integrated rotary stirring filter, the upper part of the fixed seat is connected with a defoamer surrounding the stirring shaft, the defoamer main body is of a frame structure, and the periphery of the frame structure is coated with a wire mesh; the mesh size of the screen is 100-120 um; the round pipe has many, and each round pipe is linked together through opening the runner in the fixing base inside, and the blast pipe that communicates with the runner passes the fixing base and stretches into inside the defoamer.
As a further improvement of the integrated rotary stirring filter, the frame structure of the defoamer main body comprises a top flange, a bottom flange and a vertical plate which is connected between the top flange and the bottom flange and forms an included angle of 30-60 degrees with the stirring shaft.
As a further improvement of the integrated rotary stirring filter, the circular tube extends in the radial direction of the stirring shaft, and the end face of the circular tube far away from the axis of the stirring shaft forms an included angle of 30-60 degrees with the axis of the circular tube.
The utility model has the advantages that:
1. solving the problems of nutrient supply and mixing; the air inlet channel arranged at the central position of the stirring shaft is communicated with an external air source (the prior art). The air inlet channel is communicated with an air outlet pipe through a transition channel and a communicating pipe, air outlet holes are uniformly distributed on the air outlet pipe, and the air outlet pipe is fixed at the bottom of a cage-shaped gas exchange cavity in the cage-shaped gas exchanger. The gas inlet channel, the gas outlet pipe and the like are four gas inlet pipes: oxygen can be introduced to provide oxygen for cell growth; air can be introduced to improve the dissolved oxygen; nitrogen can be introduced to drive away the redundant oxygen; carbon dioxide may be introduced to adjust the pH required for cell culture. The air supply adopts an annular air distributor-an air outlet pipe, bubbles can be generated along the outer wall of the air outlet pipe and in a cage type air exchange cavity, the outer layer of the cage type air exchange cavity is a screen, and the aperture of the screen is 10 mu m level.
The stirred reactor through which it passes is also frequently accompanied by bubbling during operation. For providing required oxygen for cell growth, the characteristic of the gentle mixing of needs during cell culture to animal cell to the very sensitive characteristics of tympanic bulla shearing, the utility model discloses in, the inlet channel of (mixing) shaft center department, outlet duct etc. also can regard as the tympanic bulla pipe tympanic bulla in the screen cloth, the cell in the screen cloth outside not with bubble direct contact. When the cage-shaped gas exchange cavity with the screen rotates along with the stirring shaft, bubbles are separated by the screen and are not in direct contact with cells outside the screen, so that the mixing effect can be ensured, the shearing force is as small as possible, and the requirement on the growth of animal cells can be met. The outer surface of the cage-shaped gas exchange cavity is provided with a stainless steel wire mesh or a teflon mesh.
2. The cells are uniformly distributed in the tank; the stirring paddle has a great influence on the growth of the cells, and the main consideration is how to reduce the influence of the shearing force on the cells and the need of providing a stable and uniform circulation flow for the culture of the microcarriers. The utility model discloses in adopt a pipe formula paddle, the stirring rake includes that three pipes and fixing base constitute. The circular tube type blades rotate together when the stirring shaft rotates. Three round pipes are evenly welded on the fixing seat and communicated with each other through circulation in the fixing seat. The length of the circular tube is determined by the volume, the end face of the three circular tube openings (overflow openings) far away from the stirring shaft is a 45-degree oblique cut (the area of the overflow opening is increased), the three overflow openings can enable negative pressure to be formed in the hollow circular tube along with the rotation of the stirring shaft, and the culture medium in the tank can move (flow) regularly through the three circular tubes in communication. Continued movement (flow) maintains the cells evenly distributed inside the canister.
3. Eliminating bubbles; a foam breaker is arranged on the fixed seat. The defoamer is in a frame type, and the outer layer of the frame is covered with a wire mesh. When the paddle rotates along with the stirring shaft, bubbles generated by stirring are sent into the defoamer through the exhaust pipe communicated with the flow channel, and the bubbles or foams are cut and damaged through the wire mesh at the periphery of the defoamer.
The experiment proves that the utility model discloses a rotatory stirring filter of integral type is used for the cultivation of insect cell, and final cultivation density reaches 6 x 106One cell per mL, the survival rate is more than 98%.
Drawings
FIG. 1 is a front view of an integral rotary agitator filter;
FIG. 2 is a sectional view A-A of the holder of FIG. 1;
fig. 3 is a bottom view of the outlet pipe of fig. 1.
In the figure, a stirring shaft 1, a stirring paddle 2, a cage-type gas exchanger 3, a defoamer 4 and an air inlet part 5 are arranged;
the device comprises a fixed seat 21, a circular tube 22, a circular tube end surface 221, a flow channel 23 and an exhaust pipe 24;
a frame-type cylinder 33, an upper flange 31, a lower flange 32, a screen 34 and a lower end plate 35;
a frame structure 41, a screen 42, a top flange 411, a bottom flange 412, an upright plate 413;
an air inlet passage 51, a transition passage 52, a communicating pipe 53 and an air outlet pipe 54.
Detailed Description
Referring to fig. 1, the integrated rotary stirring filter for cell culture comprises a stirring shaft 1, a stirring paddle 2, a cage-type gas exchanger 3, a defoamer 4, and an air inlet part 5.
The stirring paddle 2 comprises a fixed seat 21 and three circular tubes 22 which are uniformly connected to the periphery of the fixed seat in the circumferential direction and serve as blades. Each round pipe is communicated through a flow passage 23 formed in the fixed seat, and an exhaust pipe 24 vertically communicated with the flow passage 23 penetrates through the fixed seat 21 upwards and extends into the defoamer 4. The circular tube 22 extends in the radial direction of the stirring shaft, and the end surface 221 of the circular tube far away from the axis of the stirring shaft forms an included angle of 30 degrees with the axis of the circular tube, so that the contact area is increased. Of course, the fixing base is rotated along the direction away from the end surface 221 of the circular tube (counterclockwise for fig. 2) by the driving of the stirring shaft.
The cage gas exchanger 3 is composed of a frame-type cylinder 33, an upper flange 31 and a lower flange 32 which are hermetically connected to both ends of the frame-type cylinder, a screen 34 fixed to the periphery of the frame-type cylinder, and a lower end plate 35. The mesh size of the screen 34 is 10um-25 um. The frame-type cylinder is composed of a plurality of plates and round rods. The central position of the lower flange plate 32 is provided with a large hole, the aperture of which must be larger than the outer diameter of the annular air outlet pipe 54, so that the air outlet pipe 54 can be conveniently placed into the frame-type cylinder 33. The air outlet pipe 54 is arranged at the bottom of the cage-type gas exchanger 3, so that the air can be well conveyed to a lower point in a sterile manner and is uniformly dispersed into the material along with the slow mixing of the stirring shaft. After the upper flange and the lower flange are welded and formed with the frame-type cylinder 33, all welding seams are polished and then a layer of stainless steel screen 34 is covered on the outer surface of the polished upper flange and the polished lower flange, or other materials are adopted according to the process. And a lower end plate 35 is connected to the lower part of the lower flange plate through screws in a sealing manner.
The defoamer 4 is composed of a main body frame structure 41 and a wire mesh 42 coated on the periphery of the frame structure, wherein the frame structure 41 comprises a top flange 411, a bottom flange 412 and a vertical plate 413 which is connected between the top flange and the bottom flange and forms an included angle of 30-60 degrees with the stirring shaft. The vertical plates 413 are welded with the top flange and the bottom flange into a whole, and the distance between the vertical plates is not more than 10 mm. The vertical plate can be better and can cut up bubbles more quickly. The mesh size of the wire mesh 42 is 100-120 um. The center of the top flange 411 is provided with threads to connect with the stirring shaft 1.
The air inlet part 5 comprises an air inlet channel 51 arranged in the stirring shaft along the axis of the stirring shaft, a transition channel 52 arranged in the fixed seat, a communicating pipe 53 and an air outlet pipe 54. The air inlet channel 51, the transition channel 52, the communicating pipe 53 and the air outlet pipe 54 are communicated in sequence, and the air enters the cage-type gas exchanger 3 aseptically.
The upper end of the fixed base 21 is connected with the bottom flange 412 of the frame structure 41, and the lower end is connected with the upper flange 31 of the cage-type gas exchanger 3 through bolts. The main frame structure 41 is connected to the stirring shaft 1. The lower end of the stirring shaft 1 is butted with the fixed seat 21, an air inlet channel 51 is arranged along the axial line of the stirring shaft, and the air inlet channel is communicated with the upper end of a communicating pipe 53 which passes through the upper flange 31 through a transition channel 52 arranged in the fixed seat. The lower end of the communicating pipe 53 is communicated with an air outlet pipe 54 at the bottom in the cage-type gas exchanger, and air outlet holes are uniformly distributed on the air outlet pipe, see fig. 3.
The utility model discloses a rotatory stirring filter of integral type for cell culture has mainly solved and has been used for the high-density high expression of genetic engineering, accords with the bioreactor of GMP standard and the culture tank that some biological cell cultures used etc.. The integrated rotary stirring filter can feed back process detection and control parameters to a control information processor, provides convenience for basic biological process detection and analysis, can promote cell growth and prevent cell damage, can support cells to realize high-density culture in a large-scale bioreactor, and is favorable for foam control.

Claims (4)

1. A rotatory stirring filter of integral type for cell culture, including (mixing) shaft, stirring rake, characterized by: the cage-type gas exchanger consists of a frame-type cylinder, an upper flange plate and a lower flange plate which are connected with two ends of the frame-type cylinder, and a screen fixed on the periphery of the frame-type cylinder; the mesh size of the screen is 10um-75 um; the inner bottom of the cage-type gas exchanger is provided with an air outlet pipe, air outlet holes are uniformly distributed on the air outlet pipe, and the air outlet pipe is communicated with a communicating pipe which extends upwards through the upper flange plate; the stirring paddle comprises a fixed seat and a circular tube which is connected to the periphery of the fixed seat and is used as a paddle; the upper end of the fixed seat is connected with a stirring shaft, and the lower end of the fixed seat is connected with an upper flange plate; an air inlet channel is arranged in the stirring shaft along the axis of the stirring shaft, and the air inlet channel is communicated with the communicating pipe through a transition channel passing through the fixed seat.
2. The integrated rotary agitator filter of claim 1, wherein: the upper part of the fixed seat is connected with a defoamer surrounding the stirring shaft, the defoamer main body is of a frame structure, and the periphery of the frame structure is coated with a wire mesh; the mesh size of the screen is 100-120 um; the round pipe has many, and each round pipe is linked together through opening the runner in the fixing base inside, and the blast pipe that communicates with the runner passes the fixing base and stretches into inside the defoamer.
3. The integrated rotary agitator filter of claim 2, wherein: the frame structure of the defoamer main body comprises a top flange, a bottom flange and a vertical plate which is connected between the top flange and the bottom flange and forms an included angle of 30-60 degrees with the stirring shaft.
4. The integrated rotary agitator filter of claim 1, wherein: the circular tube extends in the radial direction of the stirring shaft, and the end surface of the circular tube far away from the axis of the stirring shaft forms an included angle of 30-60 degrees with the axis of the circular tube.
CN202022278644.0U 2020-10-13 2020-10-13 Integrated rotary stirring filter for cell culture Active CN213447165U (en)

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Application Number Priority Date Filing Date Title
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112143651A (en) * 2020-10-13 2020-12-29 南京金日轻工科技发展有限公司 A rotatory stirring filter of integral type for cell culture

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112143651A (en) * 2020-10-13 2020-12-29 南京金日轻工科技发展有限公司 A rotatory stirring filter of integral type for cell culture
CN112143651B (en) * 2020-10-13 2024-04-19 南京金日轻工科技发展有限公司 Integrated rotary stirring filter for cell culture

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