CN110713926A

CN110713926A - Continuous reactor for cell culture

Info

Publication number: CN110713926A
Application number: CN201911091337.7A
Authority: CN
Inventors: 孙杨; 聂简琪; 刘俊凯; 白仲虎
Original assignee: Shenzhen Yilin Biology Co Ltd
Current assignee: Shenzhen Yilin Biology Co Ltd
Priority date: 2019-11-10
Filing date: 2019-11-10
Publication date: 2020-01-21

Abstract

The invention provides a continuous reactor for cell culture, which can solve the technical problem that cells with low cell concentration cannot be effectively cultured in a large-volume cell reactor, so that the number of reactors increased due to passage requirements is reduced, the manufacturing cost and the passage time cost are reduced, and meanwhile, the risk of contamination can be avoided. A continuous reactor for cell culture comprises a tank body and a stirrer, and is characterized in that the lower part of an inner cavity of the tank body is of a tapered structure which is gradually reduced from top to bottom, and stirring blades are respectively arranged on the lower part and the upper part of the inner cavity of the stirrer. When the initial culture volume is small, the cell is introduced into the conical structure part of the inner cavity of the tank body in cooperation with the culture medium, compared with the liquid level in the conical structure part of the traditional tank body, the liquid level is higher, the stirring blades can be soaked in the culture medium, and then the culture medium is effectively stirred and controlled. When the culture needs to be enlarged, the culture volume can be increased only by supplementing the culture medium into the tank body.

Description

Continuous reactor for cell culture

Technical Field

The invention relates to the technical field of cell culture, in particular to a continuous reactor for cell culture.

Background

The cellular immunity treatment method is to collect the human body's own immune cells, to make targeted modification, to make the number reach a certain scale through in vitro culture, then to be transfused back to the human body to kill the pathogens, cancer cells and mutated cells in the blood and tissues, to break the immune tolerance, to activate and enhance the body's immunity.

The genetically modified cell culture device for clinical cellular immunization can realize modification, screening and culture of cells, but target cells collected from a human body at first cannot be directly introduced into a reactor of the cell culture device for culture under the condition of small collection amount, and the reasons are as follows: (1) in the in vitro cell culture, the cell inoculation number has influence on the growth of the cells, the target cells are collected less, and the culture medium is added less. (2) Considering the number of cells required to be finally reached, the required volume requirement of a tank body of the corresponding cell reactor is large and is about 1-2L, (3) a thin layer can be formed on the bottom of the tank body after a small amount of target cells are matched with a culture medium with a corresponding proportion and introduced, blades of a stirrer are arranged at intervals with the bottom wall of the tank body, the thin layer of the culture medium cannot be stirred, the pH, the temperature and the DO need to be monitored in the whole culture process, and corresponding probes also have requirements on the liquid level. Therefore, a small-scale reactor and a corresponding delivery pump and pipeline are additionally arranged on the cell culture device to realize the amplification of target cells, or the amplification culture is firstly carried out outside the cell culture device and then is introduced into the reactor, the former method increases the manufacturing cost of the whole cell culture device, and the latter method increases the risk of contamination.

Disclosure of Invention

The invention provides a continuous reactor for cell culture, which can solve the technical problem that a small amount of cells cannot be effectively cultured in a large-volume cell reactor, further reduce the number of reactors and the manufacturing cost, and simultaneously avoid the risk of contamination without the external culture operation of a cell culture device.

The technical scheme is that the continuous reactor for cell culture comprises a tank body and a stirrer, and is characterized in that the lower part of an inner cavity of the tank body is of a tapered structure which is gradually reduced from top to bottom, and stirring blades are respectively arranged on the lower part and the upper part of the inner cavity of the stirrer.

Further, the lower part of the inner cavity is of a truncated cone structure which is tapered from top to bottom.

Further, the stirrer is provided with the stirring blades at the lower end of the truncated cone structure of the inner cavity.

Furthermore, the bottom wall of the tank body is provided with an opening, a sleeve part extends upwards from the opening and is communicated with the opening, a connecting support is arranged on the outer side of the sleeve part, the upper end of the connecting support is fixedly connected with a stirring shaft extending into the sleeve part through a connecting piece, the stirring blade is installed on the connecting support, and a driving unit of the stirrer is arranged on the outer side of the lower part of the tank body and is in transmission connection with the stirring shaft.

Furthermore, the connecting piece comprises a cover-shaped part arranged above the sleeve part and an inserting part extending into the sleeve part, and the upper end of the stirring shaft extends into the inserting part and is connected with the upper end of the stirring shaft through a pin.

Further, the stirring shaft is rotatably supported in the sleeve portion through a bearing.

Furthermore, the lateral wall of the tank body is provided with a vent hole.

Furthermore, the lateral wall of the tank body is provided with a sampling port, a liquid filling port and an electrode socket.

Furthermore, an opening at the upper part of the tank body is provided with an end cover.

According to the reactor, as the lower part of the inner cavity of the tank body is of the tapered structure which is gradually reduced from top to bottom, the target cells with the same volume are matched with the culture medium and are led into the tapered structure part of the inner cavity of the tank body, compared with the target cells led into the inner cavity of the cylindrical structure of the traditional tank body, the liquid level in the tapered structure part is higher, the stirring blades can be soaked in the culture medium, the corresponding probes are also soaked in the culture medium, and further effective stirring and control are carried out on the culture medium, so that the culture of a small amount of target cells in the large-volume reactor is realized; when the culture needs to be enlarged, the culture medium is only needed to be supplemented into the tank body. This scheme need not additionally to prepare small-size reactor and supporting corresponding delivery pump, pipeline, has reduced manufacturing cost, need not the outside cultivation operation of cell culture device simultaneously, avoids the microbiological contamination risk.

Drawings

FIG. 1 is a schematic sectional view of the entire reactor of the present invention.

Fig. 2 is a schematic cross-sectional view of the can body of the present invention.

Fig. 3 is a partially enlarged schematic structural view of a portion a in fig. 1.

Detailed Description

As shown in fig. 1 ~ and fig. 3, a continuous reactor for cell culture comprises a tank 1 and a stirrer, wherein an inner cavity 13 of the tank comprises an upper part 13a and a lower part 13b, the lower part 13b is a truncated cone structure which is tapered from top to bottom, the stirrer is provided with a lower stirring blade 7b at the lower part 13b of the inner cavity, in particular the lower end of the truncated cone structure, and an upper stirring blade 7a at the upper part of the inner cavity.

The diapire of jar body 1 is equipped with opening 11, upwards extend and with the sleeve part 12 of opening intercommunication from opening, the outside cover of sleeve part 12 is equipped with linking bridge 6, linking bridge 6's upper end is through connecting piece 4 and the (mixing) shaft 3 fixed connection who stretches into in the sleeve part 12, it is specific, connecting piece 4 is including setting up in the lid form portion of sleeve part 12 top and stretching into the grafting portion in the sleeve part 12, the upper end of (mixing) shaft 3 stretches into in the grafting portion and connects through round pin 5, the (mixing) shaft 3 rotates through the bearing and supports in sleeve part 12, lower part stirring vane 7b installs on linking bridge 6, the drive unit of agitator sets up in the lower part outside of jar body 1 and is connected with the transmission of (mixing) shaft 3. When the motor part of the stirrer needs to be installed at the lower side (outside, not shown) of the tank body 1 due to the limitation of the installation space, the gap between the stirring shaft 3 and the tank body 1, which may enter the culture medium, is located at the top of the sleeve instead of the bottom wall through the arrangement of the sleeve part 12, and the sleeve can be arranged higher than the liquid level, so that the entering of the culture medium can be avoided, the risk of contamination is reduced, and meanwhile, the complexity of the sealing structure can be simplified.

The upper opening of the tank body 1 is buckled with an end cover 2, the end cover 2 is provided with a liquid filling port, the side wall of the tank body 1 is provided with a vent hole 16, a sampling port, a liquid filling port 15 and an electrode socket 14, and the electrode socket 14 is used for installing a detection electrode 8, such as a pH electrode, a DO electrode and the like.

Claims

1. A continuous reactor for cell culture comprises a tank body and a stirrer, and is characterized in that the lower part of an inner cavity of the tank body is of a tapered structure which is gradually reduced from top to bottom, and stirring blades are respectively arranged on the lower part and the upper part of the inner cavity of the stirrer.

2. A continuous reactor for cell culture according to claim 1, characterized in that: the lower part of the inner cavity is of a truncated cone structure which is gradually reduced from top to bottom.

3. A continuous reactor for cell culture according to claim 1, characterized in that: the stirrer is provided with the stirring blades at the lower end of the truncated cone structure of the inner cavity.

4. A continuous reactor for cell culture according to claim 1, characterized in that: the stirring device comprises a tank body, a stirring shaft and a stirring blade, wherein the bottom wall of the tank body is provided with an opening, a sleeve part extends upwards from the opening and is communicated with the opening, the outer side of the sleeve part is provided with a connecting support, the upper end of the connecting support is fixedly connected with the stirring shaft extending into the sleeve part through a connecting piece, the stirring blade is installed on the connecting support, and a driving unit of the stirrer is arranged on the outer side of the lower part of the tank body and is in transmission connection with the stirring shaft.

5. A continuous reactor for cell culture according to claim 4, characterized in that: the connecting piece comprises a cover-shaped part arranged above the sleeve part and an inserting part extending into the sleeve part, and the upper end of the stirring shaft extends into the inserting part and is connected with the upper end of the stirring shaft through a pin.

6. A continuous reactor for cell culture according to claim 4, characterized in that: the stirring shaft is rotatably supported in the sleeve portion through a bearing.

7. A continuous reactor for cell culture according to claim 1, characterized in that: the lateral wall of the tank body is provided with a vent hole.

8. A continuous reactor for cell culture according to claim 1, characterized in that: the lateral wall of the tank body is provided with a sampling port, a liquid filling port and an electrode socket.

9. A continuous reactor for cell culture according to claim 1, characterized in that: an opening at the upper part of the tank body is provided with an end cover.