CN219930047U - Preparation system of chimeric antigen receptor cells - Google Patents
Preparation system of chimeric antigen receptor cells Download PDFInfo
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- CN219930047U CN219930047U CN202321315152.1U CN202321315152U CN219930047U CN 219930047 U CN219930047 U CN 219930047U CN 202321315152 U CN202321315152 U CN 202321315152U CN 219930047 U CN219930047 U CN 219930047U
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- 108010019670 Chimeric Antigen Receptors Proteins 0.000 title claims abstract description 19
- 210000003370 receptor cell Anatomy 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 210000004027 cell Anatomy 0.000 claims abstract description 88
- 238000000926 separation method Methods 0.000 claims abstract description 50
- 239000011324 bead Substances 0.000 claims abstract description 36
- 238000007885 magnetic separation Methods 0.000 claims abstract description 33
- 230000005389 magnetism Effects 0.000 claims description 3
- 239000007788 liquid Substances 0.000 description 15
- 239000012530 fluid Substances 0.000 description 13
- 210000001744 T-lymphocyte Anatomy 0.000 description 9
- 210000004369 blood Anatomy 0.000 description 7
- 239000008280 blood Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 238000002560 therapeutic procedure Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 102000009027 Albumins Human genes 0.000 description 4
- 108010088751 Albumins Proteins 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 230000005347 demagnetization Effects 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000002659 cell therapy Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 206010000830 Acute leukaemia Diseases 0.000 description 1
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model discloses a preparation system of chimeric antigen receptor cells, which comprises a separation device, a magnetic separation device and a culture device which are independently arranged, wherein the separation device, the magnetic separation device and the culture device are respectively provided with an input container and an output container, the output container of the separation device can be communicated with the input container of the magnetic separation device for transferring cells, the output container of the magnetic separation device can be communicated with the input container of the culture device for transferring cells, the output container of the culture device can be communicated with the input container of the separation device for transferring cells, and the output end of the culture device and/or the input end of the separation device are provided with magnetic attraction devices for adsorbing magnetic beads. The preparation system of the chimeric antigen receptor cell has the advantages of reducing the use of a set of disposable consumables, saving the single use cost, improving the equipment utilization rate and the like.
Description
Technical Field
The utility model relates to the technical field of cell separation, in particular to a preparation system of chimeric antigen receptor cells.
Background
Currently, for cancer cells, it is necessary to extract T cells from the patient's blood, then perform genetic modification to enhance their targeting and killing ability against cancer cells, and allow the T cells to be cultured and expanded in large quantities in vitro for input into the patient's body, to recognize cancer cells in vivo, and to destroy them.
The method is called CAR-T therapy, CAR therapy is first proposed by Gross equal to the last 80 years of the last century, and the advent of immune cell therapies such as LAK, TIL, CIK and the like lays a foundation for the research of CAR-T therapy. In addition to CAR-T therapies, DC-CIK, CTL, etc. have been the direction of research for immune cell therapies to date, but from the perspective of technical maturity and application prospects, the current academic and industrial focus is still CAR-T therapies. The CAR-T therapy has remarkable curative effect on acute leukemia and non-Hodgkin's lymphoma, has good targeting, killing and durability in vitro and clinical experiments, and has great application potential and development prospect.
The existing preparation process mainly comprises five steps of separation, culture, demagnetization beads and concentration, wherein white blood cells are firstly separated from blood, then T cells are separated from the white blood cells through the magnetic beads, then the T cells are added for culture and amplification, the magnetic beads in the culture and amplification are removed, and finally the liquid with the magnetic beads removed is concentrated and then can be returned to a human body. In the prior art, each step is carried out by adopting a separate device, wherein the two steps of sorting and demagnetizing beads are usually the same device, but each step uses a set of disposable consumable materials for preparing cells, so that the consumable materials are more, and the single use cost is high; in the prior art, the steps are integrated on one device, so that although only one set of consumable materials is needed, cell culture usually needs several weeks, and other steps only need a short time (about one hour), so that the utilization rate of corresponding functions of the device is low, multiple devices are needed to be purchased when multiple experiments or production is performed, and the cost of the device is high.
Disclosure of Invention
The utility model aims to solve the technical problems of overcoming the defects of the prior art and providing a chimeric antigen receptor cell preparation system which reduces the use of a set of disposable consumables, saves the single use cost and improves the utilization rate of equipment.
In order to solve the technical problems, the utility model adopts the following technical scheme:
the preparation system of chimeric antigen receptor cells comprises a separation device, a magnetic separation device and a culture device which are independently arranged, wherein the separation device, the magnetic separation device and the culture device are respectively provided with an input container and an output container, the output container of the separation device can be communicated with the input container of the magnetic separation device for transferring cells, the output container of the magnetic separation device can be communicated with the input container of the culture device for transferring cells, the output container of the culture device can be communicated with the input container of the separation device for transferring cells, and the output end of the culture device and/or the input end of the separation device are/is provided with a magnetic attraction device for adsorbing magnetic beads.
As a further improvement of the above technical scheme:
the separating apparatus comprises a rotary drive and a separating vessel arranged on the rotary drive.
The magnetic separation device comprises a magnet and a separation container arranged on the magnet.
The culture device comprises a culture chamber and a culture container arranged in the culture chamber.
Compared with the prior art, the utility model has the advantages that:
when the preparation system of the chimeric antigen receptor cells is used, blood enters the separation equipment through the input container of the separation equipment, cell fluid with target cells is separated from the blood through the separation equipment, and the cell fluid with the target cells separated is collected in the output container of the separation equipment; transferring the cell liquid with the target cells from an output container of the separation device to an input container of the magnetic separation device, coating the target cells in the cell liquid by magnetic beads in the magnetic separation device, separating the target cells by the magnetic separation device, and collecting the target cells in the output container of the magnetic separation device; transferring the target cells from an output container of the magnetic separation device to an input container of the culture device, and amplifying the target cells by the culture device to obtain a predetermined amount of target cell liquid; the target cell fluid adsorbs the magnetic beads through the magnetic attraction device, and the magnetic beads in the target cell fluid are removed; the target cell liquid from which the magnetic beads are removed is transferred to an input container of the separation apparatus, the target cells are separated by the separation apparatus, and the target cells are extracted into an output container of the separation apparatus. Firstly, the magnetic attraction device is arranged at the output end of the culture equipment and/or the input end of the separation equipment, so that the demagnetizing beads can be realized in the output process of the culture equipment and/or the input process of the separation equipment, the separation and demagnetizing beads are prevented from being carried out on the magnetic separation equipment, the use of one disposable consumable is reduced, and the single use cost is saved; second, separating equipment, magnetism sorting facilities and cultivate the independent setting of equipment, like this, the longer equipment of cultivateing of needs when cultivateing, other shorter equipment of needs can repetitious usage, has improved the utilization ratio of equipment, has practiced thrift the cost.
Drawings
FIG. 1 is a schematic diagram of the structure of a system for preparing chimeric antigen receptor cells of the present utility model.
The reference numerals in the drawings denote:
1. a separation device; 2. a magnetic sorting device; 3. a culture device.
Detailed Description
The utility model will be described in further detail with reference to the drawings and the specific examples.
As used in this disclosure and in the claims, the terms "a," "an," "the," and/or "the" are not specific to a singular, but may include a plurality, unless the context clearly dictates otherwise. The terms "first," "second," and the like, as used in this disclosure, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. Likewise, the word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect.
Embodiment one:
fig. 1 shows an embodiment of a chimeric antigen receptor cell preparation system of the present utility model, which comprises a separation device 1, a magnetic separation device 2 and a culture device 3 which are independently arranged, wherein the separation device 1, the magnetic separation device 2 and the culture device 3 are respectively provided with an input container and an output container, the output container of the separation device 1 can be communicated with the input container of the magnetic separation device 2 for transferring cells, the output container of the magnetic separation device 2 can be communicated with the input container of the culture device 3 for transferring cells, the output container of the culture device 3 can be communicated with the input container of the separation device 1 for transferring cells, and the output end of the culture device 3 and/or the input end of the separation device 1 are provided with magnetic attraction devices for adsorbing magnetic beads.
In use, blood enters the separation device 1 through the input container of the separation device 1, cell liquid with target cells is separated from the blood through the separation device 1, and the cell liquid with the target cells separated is collected in the output container of the separation device 1; transferring the cell liquid with the target cells from the output container of the separation device 1 to the input container of the magnetic separation device 2, coating the target cells in the cell liquid by magnetic beads in the magnetic separation device 2, separating the target cells by the magnetic separation device 2, and collecting the target cells in the output container of the magnetic separation device 2; transferring the target cells from the output container of the magnetic separation device 2 to the input container of the culture device 3, and amplifying the target cells by the culture device 3 to obtain a predetermined amount of target cell liquid; the target cell fluid adsorbs the magnetic beads through the magnetic attraction device, and the magnetic beads in the target cell fluid are removed; the target cell liquid from which the magnetic beads are removed is transferred to an input container of the separation apparatus 1, the target cells are separated by the separation apparatus 1, and the target cells are extracted into an output container of the separation apparatus 1.
Firstly, the magnetic attraction device is arranged at the output end of the culture equipment 3 and/or the input end of the separation equipment 1, so that the demagnetizing beads can be realized in the output process of the culture equipment 3 and/or the input process of the separation equipment 1, the separation and demagnetizing beads are prevented from being carried out on the magnetic separation equipment 2, the use of one disposable consumable is reduced, and the single use cost is saved; second, separating equipment 1, magnetism sorting equipment 2 and cultivate equipment 3 independent setting, like this, the longer equipment 3 of cultivateing that needs when cultivateing, other shorter equipment that needs can repetitious usage, has improved the utilization ratio of equipment, has practiced thrift the cost.
In this embodiment, the separating apparatus 1 comprises a rotary drive and a separating vessel arranged on the rotary drive.
In this embodiment, the magnetic sorting apparatus 2 includes a magnet and a sorting container provided on the magnet.
In this embodiment, the culture apparatus 3 includes a culture chamber and a culture container provided in the culture chamber.
The magnetic attraction device comprises a demagnetization container and a magnet component, wherein the cell fluid passes through the demagnetization container, and magnetic beads in the cell fluid are attracted by the magnet component.
Embodiment two:
a method for preparing chimeric antigen receptor cells, which is carried out by adopting the preparation system of chimeric antigen receptor cells in the embodiment I, comprises the following steps:
s1, separating: separating the cell fluid with the target cells from the blood by the separating device 1;
s2, sorting: coating target cells in the cell sap by magnetic beads, and separating the target cells by a magnetic separation device 2;
s3, culturing: amplifying target cells by a culture device 3 to obtain a predetermined amount of target cell sap;
s4, demagnetizing beads: the target cell fluid adsorbs the magnetic beads through the magnetic attraction device, and the magnetic beads in the target cell fluid are removed;
s5, concentrating preparation: the target cells are separated by the separation apparatus 1, and the target cells are extracted.
Firstly, the target cell sap adsorbs magnetic beads through a magnetic attraction device arranged at the output end of the culture equipment 3 and/or the input end of the separation equipment 1, so that the magnetic beads in the target cell sap are removed, the demagnetization beads are prevented from being carried out on the magnetic separation equipment 2, the use of one set of disposable consumables is reduced, and the single use cost is saved; secondly, the preparation method of the chimeric antigen receptor cells is carried out by adopting the separation equipment 1, the magnetic separation equipment 2 and the culture equipment 3 which are independently arranged, and the separation equipment 1 is used for multiple times when the culture equipment 3 with longer time is used for culture, so that the utilization rate of the equipment is improved, and the cost is saved.
In this example, after S2, chimeric antigen receptor is added to the selected target cells, so that the target cells can recognize the designated cells, and then S3 is performed. If the target cell is a T cell, the T cell recognizes the cancer cell after adding the chimeric antigen receptor to the T cell.
In this example, the beads carry an activating substance that causes the target cells to divide.
In this embodiment, in S5, albumin is added to the separation apparatus 1. The albumin realizes layering in the process of separating the target cell liquid, such as supernatant, target cells and albumin from top to bottom, and the albumin can be directly injected into human body and is harmless to human body.
The target cells were isolated by the separation apparatus 1 for cell concentration, and the cell culture liquid was discharged (2L of liquid was cultured and could not be directly injected into human body). The cell fluid with target cells (such as T cells) is led from the culture device 3 (such as a polyhedral culture bin) to an output container (such as a soft bag) of the culture device 3 through a pipeline, and then led from the output container to the separation device 1 (such as a centrifugal machine) through a pipeline, wherein a magnetic attraction device for adsorbing magnetic beads is arranged at the output end of the culture device 3 and/or the input end of the separation device 1 and is used for adsorbing the cells with the magnetic beads in the cell fluid (the magnetic attraction device adsorbs the magnetic beads, and the magnetic beads cannot be directly injected into a human body).
The target cells (T cells) obtained were divided into two parts, one for examination and one for preservation. Since it is not possible to detect whether the target cells obtained have magnetic beads, it is necessary to take out a sample for detecting whether the magnetic bead content is acceptable. And if the sample to be inspected is qualified, taking out the target cells from the liquid nitrogen freezing machine, gradually heating the target cells by adopting a cell recovery instrument, and carrying out clinical application after the activity of the target cells is recovered.
While the utility model has been described in terms of preferred embodiments, it is not intended to be limiting. Many possible variations and modifications of the disclosed technology can be made by anyone skilled in the art, or equivalent embodiments with equivalent variations can be made, without departing from the scope of the utility model. Therefore, any simple modification, equivalent variation and modification of the above embodiments according to the technical substance of the present utility model shall fall within the scope of the technical solution of the present utility model.
Claims (4)
1. A system for preparing chimeric antigen receptor cells, characterized in that: including separating equipment (1), magnetic separation equipment (2) and the cultivate equipment (3) that independently set up, separating equipment (1), magnetic separation equipment (2) and cultivate equipment (3) all are equipped with input container, output container, the output container of separating equipment (1) can be used for transporting the cell with the input container intercommunication of magnetic separation equipment (2), the output container of magnetic separation equipment (2) can be used for transporting the cell with the input container intercommunication of cultivateing equipment (3), the output container of cultivateing equipment (3) can be used for transporting the cell with the input container intercommunication of separating equipment (1), the output of cultivateing equipment (3) and/or the input of separating equipment (1) are equipped with the magnetism that is used for adsorbing the magnetic bead and inhale the device.
2. The system for preparing chimeric antigen receptor cells according to claim 1, wherein: the separating apparatus (1) comprises a rotary drive and a separating vessel arranged on the rotary drive.
3. The system for preparing chimeric antigen receptor cells according to claim 1, wherein: the magnetic separation device (2) comprises a magnet and a separation container arranged on the magnet.
4. A preparation system of chimeric antigen receptor cells according to any one of claims 1 to 3, characterized in that: the culture device (3) comprises a culture chamber and a culture container arranged in the culture chamber.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321315152.1U CN219930047U (en) | 2023-05-26 | 2023-05-26 | Preparation system of chimeric antigen receptor cells |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321315152.1U CN219930047U (en) | 2023-05-26 | 2023-05-26 | Preparation system of chimeric antigen receptor cells |
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| Publication Number | Publication Date |
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| CN219930047U true CN219930047U (en) | 2023-10-31 |
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| CN202321315152.1U Active CN219930047U (en) | 2023-05-26 | 2023-05-26 | Preparation system of chimeric antigen receptor cells |
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2023
- 2023-05-26 CN CN202321315152.1U patent/CN219930047U/en active Active
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