CN211005428U - Closed cell enlarging culture device - Google Patents
Closed cell enlarging culture device Download PDFInfo
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- CN211005428U CN211005428U CN201920575777.9U CN201920575777U CN211005428U CN 211005428 U CN211005428 U CN 211005428U CN 201920575777 U CN201920575777 U CN 201920575777U CN 211005428 U CN211005428 U CN 211005428U
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Abstract
The utility model discloses a closed cell expanding culture device, which comprises a primary culture tank, a digestion tank, a secondary culture tank, a first conveying pipeline and a second conveying pipeline, wherein a liquid outlet at the bottom of the primary culture tank is communicated with a liquid inlet at the top of the digestion tank through the first conveying pipeline, and a liquid outlet at the bottom of the digestion tank is communicated with a liquid inlet at the top of the secondary culture tank through the second conveying pipeline; the digestion tank comprises a tank body, a stirring mechanism and a filtering mechanism, a waste liquid discharge pipeline and a microcarrier recovery pipeline are arranged at the bottom of the tank body in a penetrating mode, a top cover is arranged at the top of the tank body, a cleaning liquid pipeline, a digestion liquid pipeline, a termination liquid pipeline and a blowing pipeline are arranged on the top cover in a penetrating mode, the stirring mechanism is arranged in the tank body, and the filtering mechanism is horizontally arranged between the stirring mechanism and the bottom of the tank body. The device has realized that high efficiency carries out multistage amplification culture, and the high efficiency is digested, is separated the cell from the microcarrier surface, has shortened cell digestion cycle greatly, retrieves reuse to the microcarrier.
Description
Technical Field
The utility model relates to a cell culture technical field, concretely relates to airtight type cell enlargement culture apparatus.
Background
Microcarrier culture technology (microcarrier culture technology): fine particles are used as cell carriers, and the fine particles are suspended in a culture solution by stirring to ensure that cells are propagated to form a monolayer on the surfaces of the carriers. In the process of the digestion and amplification operation, in order to remove components which affect the culture on the microcarrier and the adherent cells before the digestion and amplification, the cleaning needs to be repeated for many times, then the cells on the microcarrier are digested by digestive juice, and the cells are taken as seed cells after falling off from the microcarrier, thereby realizing the amplification transfer of the microcarrier and the adherent cells into the large-volume bioreactor.
In a common cell digestion amplification system, the operation process of adopting digestive juice to carry out digestion and transfer is complicated, each cleaning process needs to be carried out again after the microcarrier is slowly settled under the action of self gravity and then the upper culture solution is removed, the whole cleaning process is time-consuming, and the cell loss rate is high; in the digestion process, the existing device only recovers cells falling off from the microcarriers, conveys the cells to a large-volume bioreactor for further amplification culture, directly discharges the microcarriers as waste liquid along with subsequent cleaning liquid, and does not consider the recovery and reuse of the microcarriers.
SUMMERY OF THE UTILITY MODEL
In order to solve the technical problem that exists among the above-mentioned background art, the utility model aims to provide an airtight type cell enlargement culture apparatus has realized that high efficiency carries out multistage enlargement culture, has improved production efficiency, and the efficient is digested, is separated the cell from the microcarrier surface, has shortened cell digestion cycle greatly to can retrieve reuse to the microcarrier.
In order to achieve the above purpose, the utility model adopts the following technical scheme:
the utility model provides a closed cell expanding culture device, which comprises a primary culture tank, a digestion tank, a secondary culture tank, a first conveying pipeline and a second conveying pipeline, wherein a liquid outlet at the bottom of the primary culture tank is communicated with a liquid inlet at the top of the digestion tank through the first conveying pipeline, and a liquid outlet at the bottom of the digestion tank is communicated with a liquid inlet at the top of the secondary culture tank through the second conveying pipeline;
the digestion tank comprises a tank body, a stirring mechanism and a filtering mechanism, wherein a waste liquid discharge pipeline and a microcarrier recovery pipeline are arranged at the bottom of the tank body in a penetrating manner, a top cover is arranged at the top of the tank body, a cleaning liquid pipeline, a digestion liquid pipeline, a termination liquid pipeline and a blowing pipeline are arranged on the top cover in a penetrating manner, the stirring mechanism is arranged in the tank body, and the filtering mechanism is horizontally arranged between the stirring mechanism and the bottom of the tank body;
the filtering mechanism comprises an annular fixed plate, a screen frame, a screen, a vibrator and a damping spring, wherein the outer side of the annular fixed plate is fixed on the inner side wall of the tank body, a circle of annular groove is arranged on the inner side of the annular fixed plate, part of the screen frame is clamped in the annular groove, the screen frame can move up and down in the annular groove, the screen is fixed on the inner side of the screen frame, the aperture of the screen is larger than the diameter of cells and smaller than the diameter of microcarriers, the vibrator is positioned between the bottom in the annular groove and the screen frame, the vibrator is fixedly connected with the bottom of the screen frame, the damping spring is arranged between the top in the annular groove and the screen frame, one end of the damping spring is fixed on the top in the annular groove, the other end of the spring is fixed on the top of the, the screen is provided with a central hole, and the central hole is communicated with the microcarrier recovery pipeline through a hose.
The improvement is that the screen is conical, the bottom of the screen faces the microcarrier recovery pipeline, the central hole is formed in the bottom of the screen, and the conical screen is convenient for recovering and utilizing microcarriers after cells are separated through the central hole and the microcarrier recovery pipeline.
The improvement lies in that, rabbling mechanism includes motor, (mixing) shaft and stirring paddle leaf, the motor is fixed to be set up on the top cap top, (mixing) shaft one end is run through the top cap with the motor links to each other, the (mixing) shaft other end extends to being close to in screen cloth department, the (mixing) shaft is close to screen cloth department is provided with multiunit stirring paddle leaf, stirring that can be abundant in washing and digestion process on the one hand makes washing liquid and digestive juice distribute evenly, improves the cleaning efficiency to realized cell homogeneous digestion, on the other hand is close to in the stirring paddle leaf of screen cloth department can effectually with the microcarrier piece or the cell piece dispersion of deposiing on the screen cloth, the cell digestion is complete, the separation effect is good, the recycle ratio of microcarrier is high.
The further improvement is that the stirring blade is provided with a through hole so as to reduce the damage to cells.
The further improvement is that a primary stirrer is arranged in the primary culture tank, a secondary stirrer is arranged in the secondary culture tank, and the microcarrier is suspended in the culture solution through stirring, so that the cells can be conveniently subcultured and differentiated on the surface of the microcarrier.
The improved structure is characterized in that a jacket is arranged on the outer side of the tank body, a water inlet is formed in the lower portion of the jacket, and a water outlet is formed in the upper portion of the jacket, so that the temperature of liquid in the tank body can be quickly and accurately adjusted, and the liquid in the tank body can be effectively kept warm.
The further improvement is that valves are arranged on the first conveying pipeline, the second conveying pipeline, the waste liquid discharge pipeline, the microcarrier recovery pipeline, the cleaning liquid pipeline, the digestion liquid pipeline, the termination liquid pipeline and the blow-beating pipeline.
Compared with the prior art, the utility model discloses following beneficial effect has:
the utility model discloses a be provided with filtering mechanism in the digestion tank, the screen cloth among the filtering mechanism can be effectual withhold the microcarrier that has the cell on the surface in the cleaning process, the quick emission of washing liquid has been realized, whole cleaning process is high-efficient quick, and the cell loss rate is low, can digest, separate the cell from the microcarrier surface high-efficiently quick in the digestion process, the cell after the separation directly gets into secondary culture tank through the second conveying pipeline and carries out the enlargement culture, the microcarrier can get into the recovery pipeline through the centre bore on the screen cloth and retrieve reuse; the screen cloth is at the annular groove internal stability up-and-down motion of annular fixed plate inboard under electromagnetic shaker and damping spring's effect among the filtering mechanism, the effectual screen cloth department of having avoided appears blocking phenomenon to can drive the microcarrier lump or the upwards vibrations of cell lump of deposit on the screen cloth, disperse under stirring paddle leaf's effect, the cell digestion is complete, the separation effect is good, and can concentrate the microcarrier of holding back on the screen cloth to central hole department fast, the recycle rate of microcarrier is high.
The utility model provides an airtight type cell enlarges culture apparatus has realized that high efficiency carries out multistage enlargement culture, has improved production efficiency, and the efficient is digested, is separated the cell from the microcarrier surface, has shortened cell digestion cycle greatly to can retrieve reuse to the microcarrier.
Drawings
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
FIG. 1 is a view showing the overall structure of the sealed cell culture apparatus of the present invention;
FIG. 2 is a view showing an internal structure of the sealed cell culture apparatus of the present invention;
FIG. 3 is a view showing the internal structure of the digestion tank of the present invention;
FIG. 4 is an enlarged view of a portion of FIG. 3 at A;
FIG. 5 is a perspective view of the digestion tank of the present invention;
FIG. 6 is another structural view of the filter mechanism of the present invention;
wherein the specific reference numerals are as follows: the device comprises a primary culture tank 1, a primary stirrer 2, a digestion tank 3, a tank body 4, a top cover 5, a cleaning liquid pipeline 6, a digestion liquid pipeline 7, a termination liquid pipeline 8, a blow-beating pipeline 9, a standby port 10, a waste liquid discharge pipeline 11, a microcarrier recovery pipeline 12, a motor 13, a stirring shaft 14, a stirring blade 15, an annular fixing plate 16, an annular groove 17, a screen frame 18, a screen 19, a central hole 20, a vibrator 21, a damping spring 22, a jacket 23, a water inlet 24, a water outlet 25, a secondary culture tank 26, a secondary stirrer 27, a first conveying pipeline 28 and a second conveying pipeline 29.
Detailed Description
The embodiment of the utility model discloses an airtight type cell enlargement culture apparatus, as shown in figure 1 and figure 2, including primary culture jar 1, digestion tank 3, secondary culture jar 26, first pipeline 28 and second pipeline 29, the leakage fluid dram of primary culture jar 1 bottom communicates through the inlet at first pipeline 28 and digestion tank 3 top, and the leakage fluid dram of digestion tank 3 bottom passes through the inlet at second pipeline 29 and secondary culture jar 26 top and communicates. Be provided with primary agitator 2 in the primary culture tank 1, primary agitator 2 one end is fixed at the top of primary culture tank 1, is provided with secondary agitator 27 in the secondary culture tank 26, and secondary agitator 27 one end is fixed at the top of secondary culture tank 26, makes the microcarrier suspension in the culture solution through the stirring, is convenient for the cell at the microcarrier surface subculture differentiation. As shown in fig. 3, the digestion tank 3 comprises a tank body 4, a stirring mechanism and a filtering mechanism, a waste liquid discharge pipeline 11 and a microcarrier recovery pipeline 12 are arranged at the bottom of the tank body 4 in a penetrating manner, a top cover 5 is arranged at the top of the tank body 4, a cleaning liquid pipeline 6, a digestion liquid pipeline 7, a termination liquid pipeline 8 and a blowing pipeline 9 are arranged on the top cover 5 in a penetrating manner, and as shown in fig. 5, a plurality of standby ports 10 are further formed in the top cover 5 and used for installing a temperature sensor, a carbon dioxide sensor, a dissolved oxygen sensor, a pH sensor and the like. The stirring mechanism is arranged in the tank body 4, and the filtering mechanism is horizontally arranged between the stirring mechanism and the bottom of the tank body 4. Valves are arranged on the first conveying pipeline 28, the second conveying pipeline 29, the waste liquid discharge pipeline 11, the microcarrier recovery pipeline 12, the cleaning liquid pipeline 6, the digestive juice pipeline 7, the termination liquid pipeline 8 and the blow-beating pipeline 9.
As shown in fig. 4, the filtering mechanism includes an annular fixing plate 16, a screen frame 18, a screen 19, a vibrator 21 and a damping spring 22, the outer side of the annular fixing plate 16 is fixed on the inner side wall of the tank 4, the inner side of the annular fixing plate 16 is provided with a ring of annular grooves 17, part of the screen frame 18 is clamped in the annular grooves 17, the screen frame 18 can move up and down in the annular grooves 17, the screen 19 is fixed on the inner side of the screen frame 18, the aperture of the screen 19 is larger than the diameter of cells and smaller than the diameter of microcarriers, the vibrator 21 is located between the inner bottom of the annular grooves 17 and the screen frame 18, the vibrator 21 is fixedly connected with the bottom of the screen frame 18, the damping spring 22 is arranged between the inner top of the annular grooves 17 and the screen frame 18, one end of the damping spring 22 is fixed on the inner top of the, the central bore 20 communicates with the microcarrier recovery line 12 via a hose. In order to realize the effect of the screen 19 moving up and down smoothly, the number of the vibrators 21 is even, and the vibrators 21 are symmetrically arranged between the bottom of the annular groove 17 and the screen frame 18. The number of the damper springs 22 is the same as that of the vibrators 21, and the positions of the damper springs 22 vertically correspond to those of the vibrators 21. The screen 19 moves up and down in the annular groove 17 on the inner side of the annular fixing plate 16 under the action of the vibrator 21 and the damping spring 22, the blocking phenomenon at the screen 19 is effectively avoided, the microcarrier block masses or cell block masses deposited on the screen 19 can be driven to vibrate upwards, the microcarrier block masses or cell block masses are dispersed under the action of the stirring paddle blades 15, the cell digestion is complete, the separation effect is good, microcarriers intercepted on the screen 19 can be quickly concentrated at the central hole 20, and the recovery and utilization rate of the microcarriers is high.
Rabbling mechanism includes motor 13, (mixing) shaft 14 and stirring paddle leaf 15, motor 13 is fixed to be set up on top cover 5 top, 14 one end of (mixing) shaft runs through top cover 5 and links to each other with motor 13, (mixing) shaft 14 other end extends to and is close to 19 departments of screen cloth, (mixing) shaft 14 is close to 19 departments of screen cloth and is provided with multiunit stirring paddle leaf 15, stirring that can be abundant in washing and digestion process on the one hand, make washing liquid and digestion liquid distribute evenly, the cleaning efficiency is improved, and cell homogeneous digestion has been realized, on the other hand is close to 15 can effectually disperse the microcarrier piece or the cell piece that deposit on screen cloth 19 of stirring paddle leaf that 19 departments of screen cloth, the cell digestion is complete, the separation effect is good, the recycle. The stirring paddle 15 is provided with a through hole, and the edge of the stirring paddle 15 is streamline, so that the damage to cells is further reduced.
As shown in FIG. 6, in order to facilitate the recovery and use of the microcarriers after the detachment of the cells through the central hole 20 and the microcarrier recovery line 12, the screen 19 is tapered such that the bottom of the tapered screen 19 faces the microcarrier recovery line 12 and the central hole 20 is provided at the bottom of the tapered screen 19.
Wherein, the outer side of the tank body 4 is provided with a jacket 23, the lower part of the jacket 23 is provided with a water inlet 24, and the upper part of the jacket 23 is provided with a water outlet 25, so that the temperature of the liquid in the tank body 4 can be quickly and accurately adjusted, and the liquid in the tank body 4 can be effectively insulated.
When in use:
the cells are cultured in the primary culture tank 1 in advance for 3-5 days, when the cells grow into a compact monolayer on the microcarriers, the microcarriers with the cells and the culture medium are conveyed into the digestion tank 3 through the first conveying pipeline 28, the waste liquid discharge pipeline 11 at the bottom of the digestion tank 3 is opened, the microcarriers with the cells are retained on the screen mesh 19, and the culture medium is discharged in time through the waste liquid discharge pipeline 11.
Adding cleaning solution into the tank 4 through the cleaning solution pipeline 6, opening the water inlet 24 and the water outlet 25 on the jacket 23 at the outer side of the tank 4, maintaining the temperature at 37 ℃, keeping the amount of the cleaning solution in the tank 4 to exceed the height of the screen 19, starting the motor 13 to stir at a low speed for cleaning, stopping stirring after stirring for a certain time, discharging the cleaning solution through the waste liquid discharge pipeline 11, opening the vibrator 21 if a blockage phenomenon occurs in the process of discharging the cleaning solution, or opening the vibrator 21 all the time in the cleaning process to avoid the blockage phenomenon, and repeating the above operations for 1-3 times until the color of the culture medium does not exist in the discharged cleaning solution,
adding digestive juice into the tank body 4 through a digestive juice pipeline 7, controlling the temperature at 37 ℃, carrying out digestion by low-speed stirring, simultaneously opening a blowing pipeline 9 and a vibrator 21 to achieve the effect of quickly digesting cells, adding a stopping solution into the tank body 4 through a stopping solution pipeline 8 after the cells are completely digested, uniformly mixing, introducing the cells which are uniformly in a suspension state into a secondary culture tank 26 through a second conveying pipeline 29, carrying out amplification culture, collecting microcarriers intercepted on a screen 19 through a central hole 20 and a microcarrier recovery pipeline 12, and enabling the microcarriers to enter the primary culture tank 1 or the secondary culture tank 26 for recycling.
It is right to have used specific individual example above the utility model discloses expound, only be used for helping to understand the utility model discloses, not be used for the restriction the utility model discloses. To the technical field of the utility model technical personnel, the foundation the utility model discloses an idea can also be made a plurality of simple deductions, warp or replacement.
Claims (7)
1. The closed cell enlarged culture device is characterized by comprising a primary culture tank, a digestion tank, a secondary culture tank, a first conveying pipeline and a second conveying pipeline, wherein a liquid outlet at the bottom of the primary culture tank is communicated with a liquid inlet at the top of the digestion tank through the first conveying pipeline, and a liquid outlet at the bottom of the digestion tank is communicated with a liquid inlet at the top of the secondary culture tank through the second conveying pipeline;
the digestion tank comprises a tank body, a stirring mechanism and a filtering mechanism, wherein a waste liquid discharge pipeline and a microcarrier recovery pipeline are arranged at the bottom of the tank body in a penetrating manner, a top cover is arranged at the top of the tank body, a cleaning liquid pipeline, a digestion liquid pipeline, a termination liquid pipeline and a blowing pipeline are arranged on the top cover in a penetrating manner, the stirring mechanism is arranged in the tank body, and the filtering mechanism is horizontally arranged between the stirring mechanism and the bottom of the tank body;
the filtering mechanism comprises an annular fixed plate, a screen frame, a screen, a vibrator and a damping spring, wherein the outer side of the annular fixed plate is fixed on the inner side wall of the tank body, a circle of annular groove is arranged on the inner side of the annular fixed plate, part of the screen frame is clamped in the annular groove, the screen frame can move up and down in the annular groove, the screen is fixed on the inner side of the screen frame, the aperture of the screen is larger than the diameter of cells and smaller than the diameter of microcarriers, the vibrator is positioned between the bottom in the annular groove and the screen frame, the vibrator is fixedly connected with the bottom of the screen frame, the damping spring is arranged between the top in the annular groove and the screen frame, one end of the damping spring is fixed on the top in the annular groove, the other end of the spring is fixed on the top of the, the screen is provided with a central hole, and the central hole is communicated with the microcarrier recovery pipeline through a hose.
2. The hermetically sealed cell culture apparatus of claim 1, wherein the mesh is tapered, a bottom of the mesh faces the microcarrier recovery conduit, and the central hole is provided in the bottom of the mesh.
3. The sealed cell culture apparatus according to claim 1, wherein the stirring mechanism comprises a motor, a stirring shaft and stirring blades, the motor is fixedly disposed on the top of the top cap, one end of the stirring shaft penetrates the top cap and is connected to the motor, the other end of the stirring shaft extends to a position close to the screen, and a plurality of groups of stirring blades are disposed on the position close to the screen.
4. The hermetically sealed cell culture apparatus according to claim 3, wherein the stirring blade has a through hole formed therethrough.
5. The sealed cell culture apparatus according to claim 1, wherein a primary stirrer is provided in the primary culture tank, and a secondary stirrer is provided in the secondary culture tank.
6. The sealed cell culture apparatus according to claim 1, wherein a jacket is provided outside the tank, a water inlet is provided at a lower portion of the jacket, and a water outlet is provided at an upper portion of the jacket.
7. The hermetically sealed cell culture apparatus according to claim 1, wherein valves are provided in the first transfer line, the second transfer line, the waste liquid discharge line, the microcarrier recovery line, the cleaning liquid line, the digestion liquid line, the stop liquid line and the blow-open line.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920575777.9U CN211005428U (en) | 2019-04-25 | 2019-04-25 | Closed cell enlarging culture device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920575777.9U CN211005428U (en) | 2019-04-25 | 2019-04-25 | Closed cell enlarging culture device |
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| CN211005428U true CN211005428U (en) | 2020-07-14 |
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| CN201920575777.9U Active CN211005428U (en) | 2019-04-25 | 2019-04-25 | Closed cell enlarging culture device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112941518A (en) * | 2020-12-03 | 2021-06-11 | 深圳众意远诚环保科技有限公司 | Acid etching waste liquid recycling device |
-
2019
- 2019-04-25 CN CN201920575777.9U patent/CN211005428U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112941518A (en) * | 2020-12-03 | 2021-06-11 | 深圳众意远诚环保科技有限公司 | Acid etching waste liquid recycling device |
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