CN211005422U - CAR-T cell transfection device and CAR-T product preparation system - Google Patents
CAR-T cell transfection device and CAR-T product preparation system Download PDFInfo
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- CN211005422U CN211005422U CN201921442897.8U CN201921442897U CN211005422U CN 211005422 U CN211005422 U CN 211005422U CN 201921442897 U CN201921442897 U CN 201921442897U CN 211005422 U CN211005422 U CN 211005422U
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Abstract
Aspects of the present disclosure relate to a CAR-T cell transfection device and CAR-T product preparation system. A CAR-T cell transfection device comprising: the culture container and the centrifuge are hermetically connected with the culture container through a centrifugal connecting pipe; further comprising: a sorting sample connecting pipe, a virus connecting pipe, a waste liquid connecting pipe and a culture medium connecting pipe which are respectively connected with the culture container in a sealing way through sealing parts. A CAR-T product preparation system comprises the CAR-T cell transfection device. The technical scheme provided by the embodiment of the disclosure can have the following beneficial effects: the device is internally communicated in a closed manner, so that the pollution risk is reduced, the cost is reduced, and the utilization rate and the production efficiency of equipment are improved.
Description
Technical Field
The present invention relates generally to the field of medical drug manufacturing, and more particularly to CAR-T cell transfection devices and CAR-T product manufacturing systems.
Background
At present, because the living environment and food sanitation conditions are not well protected and controlled, the incidence rate of cancers is increasing, the diseases become the most important disease affecting the health of people, and the cure rate is low, so that the survival of human beings is seriously threatened. With the development of medical science, the treatment of cancer is more and more important in the world, and cancer treatment methods, medicines and equipment are also greatly developed. One of the most accepted effective methods for treating cancer in the world is targeted therapy, which is a therapeutic approach at the cellular molecular level to the well-defined oncogenic site. The corresponding therapeutic drugs enter the body and specifically select carcinogenic sites to combine to take effect, so that tumor cells are specifically killed without affecting normal tissue cells around the tumor, and the molecular targeted therapy is also called biological missile. CAR-T Cell therapy, known as Chimeric Antigen Receptor T-Cell Immunotherapy. The novel precise targeted therapy for treating the tumor has a good effect on clinical tumor treatment by optimizing and improving in recent years, and is a novel tumor immunotherapy method which has a very promising effect, can be used for precisely, quickly and efficiently treating the cancer and is possible to cure the cancer.
The conventional CAR-T preparation devices have poor sealing effect, thereby causing higher pollution risk. CAR-T preparation facilities automation platform as an organic whole in the existing market, but the cost is expensive, and whole production is concentrated on an equipment and is accomplished, and in long-time cell culture process, other functions of equipment can not be used, has caused the problem that equipment utilization is low, the input-output ratio is low.
SUMMERY OF THE UTILITY MODEL
In order to solve the above-mentioned problems occurring in the prior art, the present invention provides a CAR-T cell transfection device, comprising:
a culture vessel for transfection of the sorted CAR-T cells;
the centrifuge is hermetically connected with the culture container through a centrifugal connecting pipe;
further comprising: a sorted sample connecting pipe, a virus connecting pipe, a waste liquid connecting pipe and a culture medium connecting pipe which are respectively connected with the culture container in a sealing way;
the other ends of the sample connecting pipe, the virus connecting pipe, the waste liquid connecting pipe and the culture medium connecting pipe, which are connected with the centrifuge, are sealed through sealing parts.
In one example, the method further comprises:
and the waste liquid container is hermetically connected with the centrifuge through a waste liquid connecting pipe.
In one example, the waste container is a flexible container.
In one example, the culture container is a flexible container.
In one example, the method further comprises:
a culture chamber, wherein the culture container is arranged in the culture chamber, the culture chamber can be filled with CO2, the temperature of the culture chamber can be adjusted, and a vibrator is arranged in the culture chamber.
In one example, the culture container is provided with a first air filter.
In one example, the centrifuge is provided with a second air filter.
In one example, the second air filter is provided with an air pressure detection device.
In one example, the method further comprises:
and an auxiliary reagent connecting tube hermetically connected to the culture container, and the other end of the auxiliary reagent connecting tube, which is opposite to the end connected to the culture container, is sealed by a sealing member.
A CAR-T product preparation system comprising:
the CAR-T cell transfection device described above.
The technical scheme provided by the embodiment of the disclosure can have the following beneficial effects: the device is internally communicated in a closed manner, so that the pollution risk is reduced, the cost is reduced, and the utilization rate and the production efficiency of equipment are improved.
Drawings
The above and other objects, features and advantages of embodiments of the present invention will become readily apparent from the following detailed description read in conjunction with the accompanying drawings. Several embodiments of the present invention are illustrated by way of example, and not by way of limitation, in the figures of the accompanying drawings and in which:
figure 1 shows a schematic structural view of a CAR-T cell transfection device as set forth in embodiments of the present invention.
In the drawings, the same or corresponding reference numerals indicate the same or corresponding parts.
Detailed Description
The principles and spirit of the present invention will be described with reference to a number of exemplary embodiments. It should be understood that these embodiments are given solely for the purpose of enabling those skilled in the art to better understand and thereby implement the present invention, and are not intended to limit the scope of the invention in any way.
It should be noted that although the terms "first", "second", etc. are used herein to describe various modules, steps, data, etc. of embodiments of the present invention, the terms "first", "second", etc. are used merely to distinguish between various modules, steps, data, etc. and do not indicate a particular order or degree of importance. Indeed, the terms "first," "second," and the like are fully interchangeable.
Figure 1 shows a schematic structural view of a CAR-T cell transfection device as set forth in embodiments of the present invention. As shown in figure 1, a CAR-T cell transfection device comprising: culture vessel 1 for transfection of sorted CAR-T cells; a centrifuge 2 hermetically connected to the culture container 1 through a centrifugal connection pipe 21; further comprising: a sorted sample connecting pipe 3, a virus connecting pipe 4, a waste liquid connecting pipe 71 and a culture medium connecting pipe 6 which are hermetically connected with the culture container 1 respectively; the other ends of the sample connection tube 3, the virus connection tube 4, the waste liquid connection tube 71 and the medium connection tube 6, which are connected to the centrifuge 1, are sealed by a sealing member 8.
The above structure constitutes a modular device for the cell transfection process stage of the CAR-T product production system. It should be noted that the above-mentioned structure must be assembled and formed in a sterile environment. The method comprises the steps of firstly, hermetically connecting a centrifuge and a culture container through a centrifugal connecting pipe in an aseptic environment, hermetically connecting a sorted sample connecting pipe, a virus connecting pipe, an auxiliary reagent connecting pipe, a waste liquid connecting pipe and a culture medium connecting pipe with the culture container respectively, and then sealing the other ends of the connecting pipes through sealing parts. Constitute the overall structure of the CAR-T cell transfection device. The sealing means may be a band, etc., and in this embodiment, a medical adhesive is used for sealing. The medical adhesive member may be an adhesive tape, an adhesive, or the like made of a medical adhesive material commonly used in the medical field.
In use, the sorted sample prepared in the previous step is input into the culture container through the sample connecting pipe, and O is mixed in a specified ratio2And CO2In the environment of (1), the culture container is shaken under the condition of specified temperature, and after the culture is carried out for specified time, whether the sample is suitable for virus transfection is detected, if the transfection condition is met, a specified amount of virus vector is input into the culture container through the virus connecting pipe, and the sample in the culture container is subjected to gene modification.
Genetic modification is one of the gene therapy mechanisms. The gene modification mainly refers to modifying a DNA sequence by a biochemical method, introducing a target gene segment into a host cell, or deleting a specific gene segment from a genome, so as to achieve the effect of changing the host cell genotype or enhancing the original genotype. Genetic modification is widely applied to various fields of human life, for example, in medicine, a genetic modification method can be used for inhibiting virus replication in certain virus host cells so as to achieve the aim of treatment.
During or after the culture, if necessary, the cultured cells in the culture container can be transferred to a centrifuge through a centrifugal connecting pipe for centrifugation, the supernatant can be discharged through a waste liquid connecting pipe, then fresh culture medium is transferred to the centrifuge through a culture liquid connecting pipe for suspending the cells, and then the prepared liquid is transferred to the culture container for continuous culture. Finally, after the culture for a specified time, obtaining a cell transfection object, disconnecting the culture container from each connecting pipe in an aseptic environment, sealing the opening for sample storage, or hermetically connecting with the next procedure for the preparation process of the next procedure.
Through above-mentioned CAR-T cell transfection device, the inside airtight intercommunication of device has reduced the pollution risk, and simple structure, used each container and pipeline all can adopt daily medical, for example plastic infusion bag, glass container, transfer line etc. the cost is reduced, and the transfection operation can independently be gone on, goes on at the transfection technology, does not influence the operation of other process, has improved the utilization ratio and the production efficiency of equipment.
In one example, the method further comprises: the waste liquid container 7 is hermetically connected to the centrifuge 1 through a waste liquid connection pipe 71.
In this embodiment, in order to facilitate the use of the above-mentioned device, a waste liquid container hermetically connected to the culture container through a waste liquid connection pipe may be provided in the device, so that the waste liquid can be discharged to the waste liquid container at any time, thereby avoiding the pollution of the waste liquid to the surrounding environment.
In one example, the waste liquid container 7 is a flexible container.
In order to reduce the cost and facilitate the installation and use, in this embodiment, the waste liquid container is a flexible container, such as a medical infusion bag.
In one example, the culture container 1 is a flexible container.
In order to reduce the cost and facilitate the installation and use, in this embodiment, the culture container is a flexible container, such as a medical infusion bag. Besides, other containers of the device can be selected from one of plastic bags, glass bottles, metal containers and the like according to specific situations.
In one example, the apparatus further comprises: a culture chamber (not shown) in which the culture vessel 1 is installed and which can be filled with CO2And the temperature is adjustable, and a shaker (not shown) is provided in the culture chamber.
In order to facilitate the transfection culture process of the cultured sample, a special culture chamber can be provided, the culture container is placed in the culture chamber, and the culture chamber is filled with CO2Can be prepared by charging CO2The container(s) is (are) connected to the culture chamber, or made as one body, or can be used for preparing CO2The device and the culture are communicated with each other by conventional technical means, and the like, and are not described in detail herein. When in use, the culture chamber can adjust the CO filled in2And O2In such a ratio that CO2Plays a role of culture room gasThe function of body partial pressure, thus meet the air pressure condition of the cultured sample; the temperature in the culture chamber can be adjusted and kept constant. After the air pressure and the temperature are adjusted, the oscillator is started to oscillate the culture container. After a specified time, the operation is stopped.
In one example, the culture container 1 is provided with a first air filter 11.
In order to circulate the fluid in the closed apparatus, it is necessary to control the input and output of the gas in the closed apparatus, and therefore, it is necessary to provide a gas passage in the culture container, and an air filter is provided in the air passage in order to ensure the purity of the gas introduced into the culture container.
In one example, the centrifuge 2 is provided with a second air filter 22.
In order to circulate a fluid in a closed device, it is necessary to control the input and output of the gas in the closed device, and therefore, a gas passage needs to be provided in the centrifuge, and an air filter needs to be provided in the air passage in order to ensure the purity of the gas entering the centrifuge.
In one example, the second air filter 22 is provided with an air pressure detection device (not shown).
In order to monitor the pressure of the gas input into the centrifuge, a gas pressure monitor, such as a gas pressure gauge, is disposed on the gas channel of the centrifuge, but the gas pressure gauge is heavy and cannot realize automatic monitoring.
In one example, the method further comprises: the auxiliary reagent connecting tube 5 is hermetically connected to the culture container 1, and the other end of the auxiliary reagent connecting tube 5 connected to the culture container 1 is sealed by a seal 8.
When the test sample is suitable for virus transfection, an auxiliary reagent is input through the auxiliary reagent connecting pipe according to the preparation condition while inputting a specified amount of the virus vector into the culture container through the virus connecting pipe, and the sample in the culture container is subjected to gene modification. Effectively improves the effect of carrying out gene modification on the sample.
A CAR-T product preparation system comprising: the CAR-T cell transfection device described above.
The CAR-T cell transfection device is applied to a CAR-T product preparation system and used as a modular device of a cell transfection process stage of the CAR-T product preparation system, when the CAR-T cell transfection device is used, the whole module is conveniently connected with devices of adjacent process stages, and the production efficiency is effectively improved. And the whole module device is connected with other equipment as a whole module after being hermetically connected in a sterile environment, so that the pollution in the CAR-T product preparation process is effectively prevented. The various containers, connecting pipelines and other components can adopt conventional low-cost components, and compared with the existing integrated automatic equipment, the cost of the whole device is greatly reduced. Through the utility model discloses a CAR-T cell transfection device is carrying out the in-process of CAR-T cell transfection operation, does not influence the preparation of other products etc. and the utility model discloses a CAR-T cell transfection device repeatedly usable has improved the utilization ratio of device effectively.
Although operations are depicted in the drawings in a particular order, this should not be understood as requiring that such operations be performed in the particular order shown or in serial order, or that all illustrated operations be performed, to achieve desirable results. In certain environments, multitasking and parallel processing may be advantageous.
The methods and apparatus of the present invention can be accomplished using standard programming techniques, with rule-based logic or other logic to accomplish the various method steps. It should also be noted that the words "means" and "module," as used herein and in the claims, is intended to encompass implementations using one or more lines of software code, and/or hardware implementations, and/or equipment for receiving inputs.
Any of the steps, operations, or procedures described herein may be performed or implemented using one or more hardware or software modules, alone or in combination with other devices. In one embodiment, the software modules are implemented using a computer program product comprising a computer readable medium containing computer program code, which is executable by a computer processor for performing any or all of the described steps, operations, or procedures.
The foregoing description of the implementation of the invention has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form disclosed, and modifications and variations are possible in light of the above teachings or may be acquired from practice of the invention. The embodiments were chosen and described in order to explain the principles of the invention and its practical application to enable one skilled in the art to utilize the invention in various embodiments and with various modifications as are suited to the particular use contemplated.
Claims (10)
1. A CAR-T cell transfection device comprising:
a culture vessel for transfection of the sorted CAR-T cells;
the centrifuge is hermetically connected with the culture container through a centrifugal connecting pipe;
further comprising: a sorted sample connecting pipe, a virus connecting pipe, a waste liquid connecting pipe and a culture medium connecting pipe which are respectively connected with the culture container in a sealing way;
the sample connecting pipe, the virus connecting pipe, the waste liquid connecting pipe and the culture medium connecting pipe are sealed by a sealing element at the other end connected with the centrifuge.
2. The CAR-T cell transfection device of claim 1, further comprising:
and the waste liquid container is hermetically connected with the centrifuge through the waste liquid connecting pipe.
3. The CAR-T cell transfection device of claim 2,
the waste liquid container is a flexible container.
4. The CAR-T cell transfection device of claim 3,
the culture container is a flexible container.
5. The CAR-T cell transfection device of claim 4, further comprising:
a culture chamber in which the culture vessel is disposed, the culture chamber being chargeable with CO2And the temperature is adjustable, and a shaker is arranged in the culture chamber.
6. The CAR-T cell transfection device of claim 5,
the culture vessel is provided with a first air filter.
7. The CAR-T cell transfection device of claim 1,
the centrifuge is provided with a second air filter.
8. The CAR-T cell transfection device of claim 7,
the second air filter is provided with an air pressure detection device.
9. The CAR-T cell transfection device of claim 1, further comprising:
and an auxiliary reagent connecting tube hermetically connected to the culture container and sealing the other end of the auxiliary reagent connecting tube connected to the culture container by a sealing member.
10. A CAR-T product manufacturing system, comprising:
the CAR-T cell transfection device of any one of claims 1-9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921442897.8U CN211005422U (en) | 2019-08-30 | 2019-08-30 | CAR-T cell transfection device and CAR-T product preparation system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921442897.8U CN211005422U (en) | 2019-08-30 | 2019-08-30 | CAR-T cell transfection device and CAR-T product preparation system |
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| Publication Number | Publication Date |
|---|---|
| CN211005422U true CN211005422U (en) | 2020-07-14 |
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| CN201921442897.8U Active CN211005422U (en) | 2019-08-30 | 2019-08-30 | CAR-T cell transfection device and CAR-T product preparation system |
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| CN (1) | CN211005422U (en) |
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