Method, device, system and medium for treating cell sample

Abstract

Provided are a method, a device, a system and a medium for treating a cell sample. The method comprises allowing a cell sample to flow into a centrifuge container; centrifuging the cell sample in the centrifuge container to obtain an initial cell solution; allowing the initial cell solution to bind to an antigen and performing magnetic separation to separate out an enriched cell solution; activating the enriched cell solution by means of inputting a modulator to obtain an activated cell solution; performing genetic modification on the activated cell solution to obtain a target cell solution; and dispensing the target cell solution into multiple preparation bags. The method, device, system and medium for treating a cell sample realize a fully-enclosed whole process of cell sample treatment integrating cell sorting, cell magnetic sorting, cell activation, cell genetic modification and cell preparation, which meets the requirements for working in a sterile environment, and can avoid external contamination of a cell sample in the treatment process. In addition, high-efficiency, standardized and large-scale production and preparation of a target cell solution are realized.

Description

Cell sample processing method, device, system and medium

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on August 22, 2023, with application number 202311065110.1, and invention name “Cell Sample Processing Method, Device, System and Medium”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application relates to the field of bioprocessing technology, and in particular to a cell sample processing method, device, system and medium. Background Art

[0003] At present, cell therapy is a key development field in the international medical frontier. Cell sorting, cell magnetic sorting, cell activation, cell genetic modification, and cell preparation are key steps in cell therapy. The inventors realized that the existing cell sorting, cell magnetic sorting, cell activation, and cell genetic modification operations are all performed manually on equipment distributed in different areas. These operations need to be performed in a sterile environment, and each step requires high-level operation by professionals. The operation threshold is high, resulting in high site and labor costs, long cycles, and extremely low efficiency.

[0004] Application Contents

[0005] The embodiments of the present application provide a cell sample processing method, device, system, and medium to solve the problems of high cost, long cycle, and low efficiency in cell therapy in the prior art.

[0006] A cell sample processing method, comprising:

[0007] The cell sample is flowed into a centrifuge container;

[0008] Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid;

[0009] performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid;

[0010] activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid;

[0011] genetically modifying the activated cell fluid to obtain target cell fluid;

[0012] The target cell solution is divided into a plurality of preparation bags.

[0013] A cell modification processing device, comprising:

[0014] A sample inflow module, used for flowing the cell sample into the centrifuge container;

[0015] a cell separation module, configured to centrifuge the cell sample in the centrifugal container to obtain an initial cell fluid;

[0016] a cell enrichment module, configured to perform antigen binding and magnetic separation on the initial cell fluid to separate enriched cell fluid;

[0017] a cell activation module, configured to activate the enriched cell fluid by inputting a regulator to obtain activated cell fluid;

[0018] A genetic modification module, used for genetically modifying the activated cell fluid to obtain a target cell fluid;

[0019] The preparation packaging module is used to package the target cell fluid into multiple preparation bags.

[0020] A cell sample processing method system includes a centrifuge container and a controller connected to the centrifuge container, wherein the controller is used to perform the following cell sample processing method:

[0021] The cell sample is flowed into a centrifuge container;

[0022] Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid;

[0023] performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid;

[0024] activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid;

[0025] genetically modifying the activated cell fluid to obtain target cell fluid;

[0026] The target cell solution is divided into a plurality of preparation bags.

[0027] One or more readable storage media storing computer-readable instructions, wherein when the computer-readable instructions are executed by one or more processors, the one or more processors perform the following steps:

[0028] The cell sample is flowed into a centrifuge container;

[0029] Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid;

[0030] performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid;

[0031] activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid;

[0032] genetically modifying the activated cell fluid to obtain target cell fluid;

[0033] The target cell solution is divided into a plurality of preparation bags.

[0034] The above-mentioned cell sample processing method, device, system and medium achieve the following: flowing the cell sample into a centrifugal container; centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid; subjecting the initial cell fluid to antigen binding and magnetic separation to separate enriched cell fluid; activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid; genetically modifying the activated cell fluid to obtain a target cell fluid; and packaging the target cell fluid into multiple preparation bags.

[0035] The present application is by centrifuging the cell sample in a centrifugal container, and realizing antigen binding and magnetic separation after centrifugation in the centrifugal container, realizing the enrichment of cell fluid, and activating the enriched cells in the centrifugal container by a regulator, and then genetically modifying the activated cell fluid to obtain the target cell fluid, and finally preparing the target cell fluid from the centrifugal container into each preparation bag, realizing a fully enclosed cell sample processing process of integrating cell sorting, cell magnetic sorting, cell activation, cell genetic modification, and cell preparation, ensuring the requirement of working in a sterile environment, and being able to avoid external contamination of the cell sample during the treatment process, and improving cell culture quality. The above steps are completed in an integrated manner, avoiding the scattered settings between equipment, reducing the occupied space, reducing manual intervention, thereby reducing site cost and labor cost. And then reducing the cycle of cell preparation, realizing the automation and standardization of target cell fluid preparation process, and improving the efficiency of the cell preparation for preparing target cell fluid.

[0036] The details of one or more embodiments of the disclosure are set forth in the accompanying drawings and the description below, and other features and advantages of the disclosure will be apparent from the description, the drawings, and the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments of the present application. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0038] FIG1 is a flow chart of a cell sample processing method according to an embodiment of the present application;

[0039] FIG2 is a flow chart of step S20 of the cell sample processing method according to one embodiment of the present application;

[0040] FIG3 is a schematic diagram of a cell sample processing device according to an embodiment of the present application;

[0041] FIG4 is a schematic diagram of a cell sample processing method system according to an embodiment of the present application. DETAILED DESCRIPTION

[0042] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0043] In one embodiment, as shown in FIG1 , a cell sample processing method is provided, comprising the following steps:

[0044] S10. Flow the cell sample into a centrifuge container.

[0045] It is understandable that initially, the cell sample is contained in one or more cell sample liquid bags, and the cell sample in the cell sample liquid bag is transported to the centrifuge container along the input pipeline by a non-contact conveying device (preferably a peristaltic pump near the liquid inlet). The non-contact conveying device can be provided in the peristaltic pump near the liquid outlet, and by generating negative pressure in the centrifuge container, the cell sample in the cell sample liquid bag flows into the centrifuge container. The input pipeline connects the cell sample liquid bag and the centrifuge container, and the opening of the cell sample flowing into the input pipeline can also be controlled by a switch valve. The input pipeline can include a filter and a dripping funnel, etc.

[0046] Furthermore, the cell sample processing method provided in this embodiment is implemented through an integrated cell sample processing device, and the entire device is placed in a sterile environment; a centrifuge container is installed on this device, which is in a sealed sterile environment, and an input pipe is connected to the centrifuge container, the other end of which can be connected to a collection bag (the collection bag is used to store solutions such as cell fluid or other reagents). In this way, it can be ensured that the cell sample is input into the centrifuge container, and the regulator and the like are input into the centrifuge container in subsequent steps are in a sealed sterile environment, and the quality of the cell fluid or the reagent itself will not be damaged by environmental influences, thereby improving the quality of cell culture.

[0047] S20, centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid.

[0048] It is understood that the cell sample is a sample comprising one or more of T cells, T cell subsets, and / or T cell progenitors. The initial cell fluid refers to a PBMC fluid or a T cell fluid. A centrifuge container is a container for holding a cell sample, including but not limited to a centrifuge cup, piston centrifuge cylinder, centrifuge tank, or centrifuge bottle.

[0049] The PBMC fluid is a cell fluid containing peripheral blood mononuclear cells (PBMCs). PBMCs refer to cells with a single nucleus in peripheral blood, including lymphocytes, monocytes, dendritic cells (DCs), and a small number of other cells. Most PBMCs are lymphocytes, including B cells and T cells.

[0050] Specifically, the cell sample is flowed into the centrifuge container in the integrated device through the liquid inlet of the centrifuge container (open when in use and closed when finished), and an appropriate sucrose polymer (e.g., Ficoll solution) can be added at the same time. The centrifuge container is controlled to perform centrifugal treatment at a preset speed. Through density gradient centrifugation, the cell sample is stratified according to density and cell size, and the red blood cell layer (mainly composed of red blood cells), the liquid layer (mainly composed of the Ficoll liquid), the PBMC layer (mainly composed of mononuclear cells) and the supernatant layer (mainly composed of plasma and blood cells) are separated. Plate), extract the intermediate product cell fluid from the liquid outlet near or located at the PBMC layer. The rotation speed can be continuously adjusted during the process to fully extract the intermediate product cell fluid in the centrifugal container. In one embodiment, the rotation speed is continuously reduced to tilt the boundaries of each layer, that is, the vertical plane is tilted, so that the boundary of the PBMC layer can cover the liquid outlet. The liquid outlet is located below the centrifugal container, so as to fully extract the intermediate product cell fluid. The intermediate product cell fluid may include liquid from the separation layer, the PBMC layer and the supernatant layer. The intermediate product cell fluid is pumped into the centrifugal container by a peristaltic pump near the liquid outlet or a peristaltic pump near the liquid inlet. The cell fluid is transferred to the intermediate product bag through the transfer pipeline, and then the cleaning fluid is introduced to clean the centrifugal container and pipeline. The flow direction of the cleaning fluid can be cleaned according to the design requirements of the pipeline. A switch valve (or a control valve) is set in the pipeline to control the opening and closing of different positions in the pipeline path through the switch valve, so that the cleaning fluid cleans each pipeline path and flushes the unnecessary components in the centrifugal container into the waste liquid bag. The cleaning fluid can be a mixture of physiological saline and 0.5% HSA (human serum albumin). After cleaning, the intermediate product in the intermediate product bag is removed by a peristaltic pump. When the product cell fluid flows back into the centrifugal container, the peristaltic pump can be reversed for transmission (for example, the peristaltic pump rotates clockwise during extraction and counterclockwise during reflow). Then, the centrifugal container is controlled to rotate centrifugally to re-stratify the intermediate product cell fluid in the centrifugal container, and a cleaning fluid is input to combine the components of the separation layer and the supernatant layer with the cleaning fluid (hydrophilic combination) to form a layer, so that the liquid of the combined layer containing the above components can be extracted from the liquid outlet to the waste liquid bag, and the rotation speed of the centrifugal container is continuously adjusted to retain the liquid of the PBMC layer, and this liquid is determined to be the initial cell fluid.

[0051] In another embodiment, non-T cells can also be labeled, and then the rotation of the centrifugal container is controlled to stratify the labeled cell sample by centrifugal force, and then extracted into an intermediate product bag. The liquid in the intermediate product bag is then returned to the centrifugal container, and the liquid is washed and then extracted. During the extraction process, filtration or pipeline adsorption is added for the labeled non-T cells to obtain an initial cell fluid containing T cells, and finally the obtained initial cell fluid is returned to the centrifugal container.

[0052] The integrated device is a device that integrates cell sorting, cell magnetic sorting, cell activation, cell genetic modification, and cell preparation. The centrifugal container includes a liquid inlet and a liquid outlet. Both the liquid inlet and the liquid outlet can be used for liquid inlet and outlet, that is, both can flow in and out of liquid. In one embodiment, the liquid inlet of the centrifugal container is located above the container, and the liquid outlet of the centrifugal container is located below the container.

[0053] S30, performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid.

[0054] It can be understood that the enriched cell fluid refers to a cell fluid enriched with T cells.

[0055] Specifically, the antigen liquid and the magnetic beads are transported to a centrifugal container, wherein the antigen liquid is a liquid containing antigen molecules, for example: antigen binding molecules CD2, CD3, CD4, CD8, CD25, CD28, CD27, CD45RA, CD45RO, CD62L, CD95, CD127, CD137, α / βTCR, γ / δTCR, CCR7, PD-1 or Lag3, etc., and the magnetic beads can be micron-sized magnetic beads (magnetic beads with a particle size of 1 to 5 microns) or nanometer-sized magnetic beads (magnetic beads with a particle size of 5-100 nanometers). During the pipeline process of transporting the antigen liquid and the magnetic beads to the centrifugal container, the antigen liquid and the magnetic beads can be flowed into the same pipeline and then passed through the liquid inlet. The liquid flows into a centrifugal container, which can rotate back and forth at a specific speed to allow the initial cell fluid to bind to antigens. The antigen binding is to bind antigen molecules and magnetic beads to cells in the initial cell fluid, and the initial cell fluid after antigen binding is extracted from the centrifugal container through a peristaltic pump near the liquid outlet to a magnetic separation column near the liquid outlet. The pipeline between the liquid outlet and the magnetic separation column can pass through the peristaltic pump, and then magnetic separation is achieved through the magnetic separation column, that is, the initial cell fluid is subjected to immunomagnetic cell separation by magnetic bead sorting, that is, the affinity of specific antibodies or other molecules on the surface of magnetic beads with cell surface molecules is used to separate specific types of immunomagnetic cells bound to magnetic beads, thereby separating enriched cell fluid. The initial cell fluid can also be labeled by optical excitation, that is, the specific binding of fluorescent markers to cell surface molecules is used to extract the specific bound initial cell fluid from the centrifugal container, and then the specifically bound cells are transported through a specific channel according to the single cell channel. The specifically bound cells can be transported through a funnel-shaped channel to narrow the channel so that the specifically bound cells emit fluorescent signals under optical excitation for sorting, and the enriched cells are separated by lasers, detection channels and filters. The laser can excite the wavelength of the labeled cells, and then the wavelength is detected by the detector and filter, and then the wavelength presented by the required fluorescent pigment is selected, and the detected cells are harvested according to different labels, thereby harvesting the enriched cell fluid.

[0056] In one embodiment, magnetic beads and antigens can be combined with T cells to magnetically label the T cells. A peristaltic pump is then used to transfer the cell fluid containing the labeled T cells from the centrifuge container to an intermediate fluid bag. The intermediate fluid bag is then moved onto a variable magnetic device. An external magnetic field is applied to the variable magnetic device, thereby adsorbing the T cells bound to the magnetic beads and antigen in the fluid of the intermediate fluid bag. The variable magnetic device is then controlled to oscillate or vibrate using a swing parameter (which can be set to swing the variable magnetic device at different angles depending on the actual situation) or a vibration parameter to uniformly distribute the T cells bound to the magnetic beads and antigen adsorbed by the variable magnetic device. This allows all substances in the intermediate fluid bag, other than the adsorbed T cells bound to the magnetic beads and antigen, to be suspended in the fluid. A waste liquid draining operation is then performed to allow the fluid in the intermediate fluid bag, other than the adsorbed T cells bound to the magnetic beads and antigen, to flow into the waste liquid bag. The T cells bound to the magnetic beads and antigen in the intermediate fluid bag are then washed. Finally, a resuspension solution or buffer is injected into the intermediate fluid bag to separate the enriched cell fluid, which is then returned to the centrifuge container. Following subsequent cell culture, magnetic bead removal can be performed to obtain the final cells. A variable magnetic device can alter the strength of the magnetic field, or the magnetic separation column can be switched between magnetic and non-magnetic by moving the magnet.

[0057] S40, activating the enriched cell fluid by inputting a regulator to obtain activated cell fluid.

[0058] It can be understood that the activated cytosol refers to the cytosol after T cells are activated.

[0059] Specifically, the regulator is placed in a regulator liquid bag, and the regulator can be injected into the centrifugal container through an inlet and outlet of the centrifugal container by a peristaltic pump. The regulator is a solvent that can activate T cells. The activation of T cells by the regulator requires the participation of a dual-signal system. For example, in addition to the first signal provided by MHC / antigen peptide-TCR (T cell antigen receptor, T cell receptor), a second signal must be provided by a costimulatory molecule to activate T cells. The regulator stimulates the T cell receptor to receive the costimulatory signal, thereby activating the T cell and obtaining an activated cell fluid. The B7 protein on the APC (Antigen-Presenting Cell, antigen presenting cell) can provide the costimulatory signal necessary for activation and produce the following costimulatory effect after binding to the CD28 receptor of the enriched cell fluid containing T cells. The B7 protein binds to the CTLA-4 (cytotoxic T lymphocyte-associated protein 4) receptor to promote the deactivation of T cells.

[0060] S50, genetically modifying the activated cell fluid to obtain target cell fluid.

[0061] Understandably, the genetic modification can be obtained by a variety of methods, such as viral and non-viral vectors, mRNA (electroporation technology), genome editing tools, etc. Among them, in the viral vector method, the retroviral vector (RV) is a widely used viral vector, and the lentiviral vector (lentiviral vector) contains a cis-acting element central polypurine tract (cPPT), which enables it to effectively infect resting T cells. The genetic modification can transfer the structure of the CAR into the activated cell fluid (activated T cells) through genetic engineering, and finally obtain the target cell fluid, which is called a targeted CAR-T (Chimeric Antigen Receptor T-Cell Immunotherapy) cell. Among them, the activated cell fluid can be genetically modified by one of viral vectors, transposons, nanoparticles, liposomes and electroporation-mediated transfection.

[0062] S60, packaging the target cell solution into multiple preparation bags.

[0063] Specifically, after the target cell fluid, the target cell fluid is divided into a preparation by a peristaltic pump and then divided into multiple pre-set preparation bags. A quantitative tube can also be introduced to accurately divide the target cell fluid into multiple preparation bags, and the preparation bags are squeezed to empty them.

[0064] In one embodiment, during the process of packaging the target cell fluid into each of the preparation bags, the centrifugal container can be rotated left and right to mix the target cell fluid so that it will not precipitate, thereby ensuring the activity of the T cells, and then packaged into each preparation bag. The packaging process is to package the liquid in the centrifugal container into a corresponding volume of each preparation bag according to the preparation requirements of the preparation bag.

[0065] All pipelines in this application can be equipped with sensors for monitoring as needed, such as bubble sensors for monitoring the direction and flow rate of liquid in the pipeline, pressure sensors for monitoring the pressure in the pipeline, flow sensors for monitoring the flow rate of transmission, photoelectric sensors for detecting the volume of liquid in the centrifugal container, etc. All pipelines in this application can be equipped with switch valves for opening and closing the pipelines as needed, such as solenoid valves for opening and closing the input and output of each liquid bag.

[0066] In one embodiment, the target cell fluid in the centrifugal container is quantitatively prepared through a quantitative tube to distribute the quantitatively prepared target cell fluid into each preparation bag. The quantitative preparation process is to respectively provide a bubble sensor / liquid sensor on the inlet pipe and the outlet pipe of the quantitative tube, and monitor the time when the liquid enters the quantitative tube and flows out of the quantitative tube through the bubble sensor / liquid sensor, which are defined as the initial time and the final time. According to the fixed capacity of the quantitative tube and the time difference between the two times, the constant flow rate of the liquid flowing through the peristaltic pump with a constant speed during this period can be determined. The constant flow rate can be accurately distributed into each preparation bag because the working time for distributing each preparation bag is obtained by dividing the capacity of the preparation bag by the constant flow rate. The time of entering the preparation bag is sensed by the sensor provided on each preparation bag, thereby improving the accuracy of the distribution amount of each preparation bag. Before the bubble sensor / liquid sensor detects the moment when the liquid enters and flows out of the quantitative tube, the liquid entering the quantitative tube can be debubbled. Initially, the inlet of the quantitative tube can be located below the outlet of the quantitative tube, and the bubbles in the quantitative tube can be emptied by the principle of rising bubbles. After the quantitative tube is filled, the quantitative tube can be set so that the inlet of the quantitative tube is higher than the outlet of the quantitative tube, so that the quantitative tube remains filled with liquid.

[0067] In the present application, by centrifuging the cell sample in a centrifugal container and realizing antigen binding and magnetic separation after centrifugation in the centrifugal container, the enrichment of the cell fluid is realized, and the enriched cells are activated in the centrifugal container by a regulator, and then the activated cell fluid is genetically modified to obtain the target cell fluid, and finally the target cell fluid preparation is transferred from the centrifugal container to each preparation bag, realizing a fully enclosed and full-flow cell sample processing process integrating cell sorting, cell magnetic sorting, cell activation, cell genetic modification, and cell preparation, ensuring the requirements of working in a sterile environment, and being able to avoid external contamination of the cell sample during the processing. And then the automation and standardization of the target cell fluid preparation process are realized, reducing site costs and labor costs, and realizing the high-efficiency, standardized, and large-scale production and preparation of the target cell fluid.

[0068] In one embodiment, as shown in FIG2 , the step S20 , i.e., centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid, includes:

[0069] S201. Control the centrifugal container to rotate at a preset speed so that the cell sample in the centrifugal container can be centrifuged and separated; the cell sample includes at least one of T cells, T cell subsets, and T cell progenitor cells.

[0070] S202, separating the initial cell fluid from the cell sample after centrifugation and stratification.

[0071] It is understood that the preset rotation speed is the rotation speed of the centrifuge container set in advance. The initial cell fluid is PBMC fluid or T cell fluid. The cell sample includes at least one of T cells, T cell subsets, and T cell progenitor cells.

[0072] Specifically, after the cell sample flows into the centrifuge container, the centrifuge container is controlled to rotate at a preset speed. Due to the different cell densities and sizes, different cells are stratified into different cell layers through centrifugation. Then, the cell layer containing the PBMC fluid or T cell fluid is separated, that is, the initial cell fluid is separated from the cell sample after centrifugation and stratification. After the cell sample has completed stable stratification, the centrifuge container is controlled to stop rotating, thereby preventing cell damage in the cell sample during excessive centrifugation. In other words, in this embodiment, separation of the initial cell fluid is achieved through centrifugation and stratification at a preset speed, thereby improving the efficiency of separating the initial cell fluid.

[0073] In another embodiment, the step S20, i.e., centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid, includes:

[0074] S203 , labeling the cells in the cell sample to obtain labeled cells and unlabeled cells.

[0075] S204 , performing centrifugation to remove labels from all the labeled cells and all the unlabeled cells to obtain the initial cell fluid.

[0076] It is understood that non-marked cells refer to T cells, T cell subsets and / or T cell progenitor cells, while marked cells refer to other cells such as B cells.

[0077] Specifically, after the cell sample flows into the centrifugal container, the cells in the cell sample are labeled, that is, a preset labeling liquid is added to the centrifugal container, and the centrifugal container is controlled to rotate so that different labeled antigens in the preset labeling liquid label different cells in the cell sample. For example, B cells can be labeled by CD19, and suppressor cells can be removed by using CD25 and CD14 respectively, so that all cells in the cell sample are divided into labeled cells and unlabeled cells. Further, all labeled cells and all unlabeled cells are centrifuged to remove the label, that is, the centrifugal container is controlled to rotate and centrifuge so that the labeled cells and all unlabeled cells are separated into layers, that is, because the labeled cells are heavier than the unlabeled cells, the two types of cells are separated by centrifugation. Then, the unlabeled cells in the specific cell layer are separated, and a small amount of labeled cells are adsorbed in the pipeline by a magnet to obtain an initial cell fluid. Among them, it is also possible to distinguish by labeling T cells, and compared with labeling other cells, labeled T cells are easier to distinguish. It is only necessary to centrifuge the labeled T cells and adsorb the labeled T cells to obtain the initial cell fluid. By labeling cells in a cell sample, T cells and non-T cells in the cell sample can be distinguished. By centrifuging the labeled cell sample, T cell fluid can be separated and the initial cell fluid can be obtained.

[0078] In one embodiment, the step S30, i.e., performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid, includes:

[0079] S301. Add magnetic beads and antigens to a centrifugal container containing the initial cell fluid, so that the magnetic beads and antigens are combined with the initial cells in the initial cell fluid to obtain a mixed cell fluid.

[0080] It can be understood that the mixed cell fluid refers to the cell fluid after the magnetic beads and antigens are coupled to the initial cells.

[0081] Specifically, after the initial cell fluid is separated, magnetic beads and antigens are added to the centrifugal container containing the initial cell fluid. The antigen is directly or indirectly coupled to the surface of the magnetic beads to form a magnetic bead complex with affinity, so that the magnetic bead complex binds to the T cells in the initial cell fluid, that is, the centrifugal container is rotated to uniformly mix the initial cell fluid and the magnetic bead complex, so that the magnetic bead complex binds to the T cells, that is, the antigen is directly or indirectly coupled to the magnetic bead complex, specifically recognized by the T cell surface antibody, and combined with the T cell surface antibody, so that the magnetic bead complex is directly connected to the cell to form a magnetic marker, thereby obtaining a mixed cell fluid.

[0082] In one embodiment, micron-sized magnetic beads and antigens are added to a centrifugal container containing an initial cell fluid. The antigen is directly or indirectly coupled to the surface of the micron-sized magnetic beads to form a micron-sized magnetic bead complex with affinity, so that the micron-sized magnetic beads bind to non-T cells in the initial cell fluid. That is, the centrifugal container is rotated to uniformly mix the initial cell fluid and the micron-sized magnetic bead complex, and the micron-sized magnetic bead complex and the non-T cells in the initial cell fluid are magnetically labeled. That is, the antigen is directly or indirectly coupled to the micron-sized magnetic bead complex, which binds to the non-T cell surface antibody to form a magnetic label for the non-T cells, thereby obtaining a micron-sized mixed cell fluid. Among them, the micron-sized magnetic beads and antigens can also be combined with T cells to achieve the acquisition of the micron-sized mixed cell fluid, and the magnetic beads need to be removed later.

[0083] In another embodiment, after separating the initial cell fluid, nanoscale magnetic beads and antigens are added to the centrifugal container containing the initial cell fluid, and the antigen is directly or indirectly coupled to the surface of the nanoscale magnetic beads to form a nanoscale magnetic bead complex with affinity, so that the nanoscale magnetic beads are combined with the T cells in the initial cell fluid, that is, the centrifugal container is rotated to uniformly mix the initial cell fluid and the nanoscale magnetic bead complex, so that the nanoscale magnetic bead complex is combined with the T cells, that is, the nanoscale magnetic bead complex is directly or indirectly coupled to the antigen, specifically recognized by the T cell surface antibody, and combined with the T cell surface antibody, so that the nanoscale magnetic beads are directly connected to the T cells to form a magnetic label, thereby obtaining a nano-mixed cell fluid. Wherein, when using nano-magnetic beads and antigens for magnetic labeling, there is no need to separate the nano-magnetic beads subsequently. Understandably, nano-magnetic beads can also be used to magnetically label non-T cells to obtain a nano-mixed cell fluid.

[0084] S302, performing magnetic bead sorting on the mixed cell fluid to separate the enriched cell fluid.

[0085] It can be understood that the enriched cell fluid refers to a cell fluid enriched with T cells.

[0086] Specifically, a peristaltic pump is used to transfer the mixed cell fluid into a fluid bag on a variable magnetic device. The variable magnetic device then applies an external magnetic field to adsorb T cells bound to magnetic beads and antigens in the fluid bag. The variable magnetic device is controlled to swing by setting appropriate swing parameters to evenly distribute the T cells bound to magnetic beads and antigens adsorbed by the variable magnetic device, preventing large accumulation of adsorbed T cells bound to magnetic beads and antigens. This also allows all substances in the fluid bag, other than the adsorbed T cells bound to magnetic beads and antigens, to be suspended in the fluid. A waste liquid discharge operation is then performed, i.e., a waste liquid discharge line connected to the fluid bag is opened to allow the fluid in the fluid bag, other than the adsorbed T cells bound to magnetic beads and antigens, to flow into the waste liquid bag through the waste liquid discharge line. Furthermore, a cleaning line connected to the fluid bag is opened to perform a cleaning operation on the T cells bound to magnetic beads and antigens in the fluid bag. Finally, a resuspension solution or buffer is injected into the fluid bag to maintain the activity of the T cells bound to magnetic beads and antigens, and the magnetic field is canceled or removed, thereby separating the enriched cell fluid.

[0087] In another embodiment, after applying an external magnetic field by a variable magnetic device to adsorb the non-T cells bound to the magnetic beads in the liquid of the liquid bag, the liquid is reversely transferred by a peristaltic pump, that is, the liquid in the liquid bag except the adsorbed non-T cells bound to the magnetic beads and antigens is input into a centrifugal container through the peristaltic pump. Then, the magnetic field is canceled or removed, the waste liquid discharge pipeline connected to the waste liquid bag is opened, and the non-T cells bound to the magnetic beads and antigens are flowed into the waste liquid bag through the waste liquid discharge pipeline. In this embodiment, since the non-T cells bound to the magnetic beads and antigens are removed, the cell fluid input into the centrifugal container by the peristaltic pump is the enriched cell fluid. That is, in this embodiment, the efficient and stable separation and collection of T cell fluid by magnetic bead sorting is achieved, and the efficient and stable capture of T cell fluid is achieved.

[0088] In one embodiment, the step S40, i.e., activating the enriched cell fluid by injecting a regulator to obtain the activated cell fluid, includes:

[0089] S401. Input the regulator into the centrifugal container containing the enriched cell fluid.

[0090] S402. Activate the enriched cells in the enriched cell fluid using the co-stimulator in the regulator to obtain the activated cell fluid; the regulator is one or more of an agonist antibody, a cytokine, a recombinant co-stimulatory molecule, and a small drug inhibitor.

[0091] It is understood that the modulator is one or more of an agonistic antibody, a cytokine, a recombinant co-stimulatory molecule, and a small drug inhibitor.

[0092] Specifically, after obtaining the enriched cell fluid, the regulator can be injected into the enriched cell fluid of the centrifugal container through an inlet and outlet in the centrifugal container by a peristaltic pump, so as to activate the cells in the enriched cell fluid, that is, the enriched cells in the enriched cell fluid are activated by the costimulator in the regulator, that is, the antigen is combined with the antigen receptor on the surface of the T cell to generate a specific signal, i.e., the first activation signal. For example, TCR / CD3 and the specific MHC (major histocompatibility complex, major tissue compatibility complex) molecule antigen peptide complex on the surface of the antigen presenting cell (APC) combine to produce a specific antigen stimulation signal. Then, the costimulator in the regulator interacts with the receptor on the surface of the T cell to produce a nonspecific costimulatory signal, which is used to activate the cells in the enriched cell fluid to obtain the activated cell fluid.

[0093] In one embodiment, the step S402, i.e., activating the enriched cells in the enriched cell fluid by the costimulator in the regulator to obtain the activated cell fluid, includes:

[0094] S4021. Using the costimulator in the regulator, the enriched cells in the enriched cell fluid produce a costimulatory effect to obtain active cell fluid.

[0095] S4022. Sampling and testing the active cell fluid to obtain a test result.

[0096] S4023. When the detection result indicates that the enriched cells have been successfully activated, the active cell fluid is determined as the activated cell fluid.

[0097] Understandably, active cell fluid refers to T cells that produce a co-stimulatory effect. The test results are used to indicate whether the enriched cells have been successfully activated.

[0098] Specifically, after obtaining the enriched cell fluid, the co-stimulator in the regulator is used to make the enriched cells in the enriched cell fluid produce a co-stimulatory effect, that is, the co-stimulator is combined with the specific cell receptor on the surface of the enriched cell to produce a specific antigen stimulation signal, and the non-specific antigen stimulation signal is generated by the interaction of multiple pairs of co-stimulatory molecules and the corresponding receptors of T cells on the surface of the enriched cell, wherein CD28 is the most important co-stimulatory molecule, and the non-specific antigen stimulation signal can fully activate T cells, secrete cytokines and express cytokine receptors, thereby obtaining active cell fluid. Further, the active cell fluid is sampled and tested, that is, a small amount of liquid (for example, 3 ml, etc.) is discharged from the discharge port of the centrifugal container into the sampling tube, that is, the cells in the sampling tube are subjected to immunological testing, and the expression level of cell surface activation-related molecules in the sampling tube, such as CD69, CD25, etc., is detected to determine whether the cells are in an activated state. It is also possible to determine whether the cells are in an activated state by detecting cytokines secreted by the cells in the sampling tube, such as IL-2, IFN-γ, etc., so that the test results can be obtained. When the test results indicate that the enriched cells have been successfully activated, the active cell fluid is determined to be the activated cell fluid. This means that the active cell fluid has been obtained. By sampling and testing the active cell fluid, the activation of the enriched cells is ensured, providing a guarantee for subsequent genetic modification of T cells.

[0099] In one embodiment, step S401 includes:

[0100] The costimulator comprises at least one of an anti-CD3 antibody or fragment and an anti-CD28 antibody or fragment.

[0101] or / and

[0102] The co-stimulator comprises a nanomatrix comprising at least one of a matrix of mobile polymer chains and a matrix connected to the mobile polymer chains. Preferably, the co-stimulator is a biodegradable nanomatrix, and the nanomatrix comprises at least one of a matrix of mobile polymer chains and a matrix connected to the mobile polymer chains. The nanomatrix has a size of 1 nm to 500 nm. The same or different matrices of mobile polymer chains are connected to anti-CD3 and anti-CD28 antibodies or fragments. If the anti-CD3 and anti-CD28 antibodies or fragments are connected to different matrices of mobile polymer chains, the nanomatrix is ​​fine-tuned to stimulate T cells.

[0103] In one embodiment, the step S50, i.e., genetically modifying the activated cell fluid to obtain the target cell fluid, includes:

[0104] S501. Perform transduction treatment on the activated cell fluid to obtain the target cell fluid.

[0105] Specifically, retroviral vectors and lentiviral vectors are commonly used for viral transduction. Retroviruses integrate viral DNA (deoxyribonucleic acid, DeoxyriboNucleic Acid) into the cellular DNA in the activated cell fluid, producing stable transfection. Lentiviral vectors effectively integrate exogenous genes or exogenous RNA (ribonucleic acid, Ribonucleic Acid) into the chromosomes of cells in the activated cell fluid. Among them, the viral vector is added to the activated cell fluid in a centrifugal container through a pipeline so that the viral vector infects the cells in the activated cell fluid, that is, the viral surface envelope protein binds to the cell receptor and enters the cell. In the cell, the RNA of the viral vector is reverse transcribed into double-stranded DNA. Then, the double-stranded DNA forms an integration complex with the integrase, and the integration complex enters the cell nucleus through the nuclear pore. Then, the integration complex binds to a specific site of the T cell DNA to form a new complex. The integrase rearranges and connects the viral DNA sequence with the T cell DNA sequence, and the integrase is released from it to embed the viral genome into the DNA of the T cell, thereby obtaining the target cell fluid.

[0106] or

[0107] S502, performing transfection treatment on the activated cell fluid to obtain the target cell fluid.

[0108] It can be understood that cell transfection refers to the introduction of exogenous DNA or RNA substances into cells, such as transposons and transfection reagents.

[0109] Specifically, after obtaining the activated cell fluid, the activated cell fluid is subjected to transfection treatment, that is, the exogenous gene is inserted into the chromosome of the cell, that is, a transfection reagent can be added to the activated cell fluid in the centrifugal container through a pipeline, and the exogenous DNA or RNA is introduced into the cell nucleus in the cell using the transfection reagent. For example, DNA and calcium chloride are mixed in phosphate-buffered saline to form a calcium phosphate-DNA precipitate, and then it is dispersed on the cells by centrifugal rotation. Calcium phosphate promotes the binding of the condensed DNA in the co-precipitate to the cell surface, and the DNA enters the cell through endocytosis. Alternatively, cationic liposome-mediated transfection, that is, a positively charged head group and a negatively charged nucleic acid form a complex through electrostatic interaction, and enter the cell through endocytosis of the cell, so as to achieve subsequent genetic modification of the cells in the activated cell fluid, thereby obtaining the target cell fluid.

[0110] or

[0111] S503 , performing an electroporation operation on the activated cell fluid to obtain the target cell fluid.

[0112] Electroporation is a cell transfection method that generates transmembrane voltage by applying an external pulsed electric field, thereby forming recoverable micropores on the cell membrane surface and introducing exogenous substances into cells.

[0113] Specifically, after obtaining the activated cell fluid, the transfection substance is added to the activated cell fluid in the centrifugal container through the pipeline, and the activated cell fluid with the added transfection substance is transferred to the electroporation cell through a peristaltic pump, and an electric field is applied to the electroporation cell to produce transient holes in the cell membrane. Alternatively, the electroporation cell can be a microfluidic chip, which is equivalent to separating various flow channels in the electroporation cell so that the cell membrane can be hit by the electric field to form holes in each flow channel, so that the transfection substance carries exogenous DNA or RNA and combines with the cells in the activated cell fluid. Then, the electroporated cells are washed and transferred to a suitable liquid bag through a peristaltic pump to achieve genetic modification of the cells in the activated cell fluid, thereby obtaining the target cell fluid.

[0114] or

[0115] S504, performing mechanical transfection treatment on the activated cell fluid to obtain the target cell fluid.

[0116] Specifically, after obtaining the activated cell fluid, the activated cell fluid is subjected to mechanical transfection treatment, that is, the activated cell fluid is transferred to the extrusion device through a peristaltic pump, and the transfection material is transferred at the same time. The extrusion device mechanically squeezes the cells, and squeezes the transfection material into the cell nuclei of the cells in the activated cell fluid, promotes the formation of the transduction complex, and realizes genetic modification of the cells in the activated cell fluid, thereby obtaining the target cell fluid.

[0117] In one embodiment, the step S501, i.e., performing transduction treatment on the activated cell fluid to obtain the target cell fluid, includes:

[0118] The activated cell fluid is transduced by a transduction enhancer to obtain the target cell fluid; the transduction enhancer includes one or more of a polycationic agent, poloxamer, poloxamer, and adhesion molecules.

[0119] It is understood that transduction-enhancing agents include one or more of polycationic agents, poloxamers, and adhesion molecules. Polycationic agents are molecules with a positive charge, such as polybrene, protamine sulfate, poly-L-lysine, and peptides with a net positive charge. Adhesion molecules include fibronectin or modified fibronectin, or protein targeting domains, such as antibodies, antibody complexes, and other molecules.

[0120] Specifically, after obtaining the activated cell fluid, the activated cell fluid is transduced using a transduction enhancer, that is, the transduction enhancer contained in the fluid bag is introduced into a centrifugal container through a pipeline, and the centrifugal container is rotated so that the transduction enhancer is in full contact with the transduction substance and the cells, thereby using the transduction enhancer to enhance the interaction between the transduction substance and the cells in the activated cell fluid, promoting the binding of the transduction substance to the cells, enabling the transduction substance to bring genetic material into the activated cells, and achieving genetic modification of the cells in the activated cell fluid, thereby obtaining the target cell fluid. Among them, polycationic agents promote the formation of cell transduction complexes by interacting with the negative charges on the cell membrane. Poloxamers can be used to prepare liposomes and other nanoparticles, and liposomes and nanoparticles can be used to transduce various molecules into cells. Poloxamers can also promote the formation of cell transduction complexes by interacting with the cell membrane. Adhesion molecules can mediate the interaction between the transduction complex and the cell membrane, promoting the transduction complex to enter the cell. That is, in this embodiment, by using a transduction enhancer, the transduction efficiency of the activated cell fluid is improved, so that the transduction substance can enter the cell more easily, and the time of cell transduction is reduced.

[0121] It should be understood that the size of the serial numbers of the steps in the above embodiments does not mean the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of this application.

[0122] In one embodiment, a cell modification processing device is provided, which corresponds to the cell modification processing method in the above-mentioned embodiment. As shown in Figure 3, the cell modification processing device includes a sample inflow module 10, a cell separation module 20, a cell enrichment module 30, a cell activation module 40, a genetic modification module 50, and a preparation packaging module 60. The functional modules are described in detail as follows:

[0123] The sample inflow module 10 is used to flow the cell sample into the centrifugal container;

[0124] The cell separation module 20 is used to centrifuge the cell sample in the centrifugal container to obtain an initial cell fluid;

[0125] The cell enrichment module 30 is used to perform antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid;

[0126] A cell activation module 40 is configured to activate the enriched cell fluid by inputting a regulator to obtain activated cell fluid;

[0127] A genetic modification module 50 is used to genetically modify the activated cell fluid to obtain a target cell fluid;

[0128] The preparation packaging module 60 is used to package the target cell fluid into multiple preparation bags.

[0129] A cell sample processing method system comprises a centrifugal container and a controller connected to the centrifugal container, wherein the controller is used to execute the cell sample processing method.

[0130] Specific definitions of the cell sample processing method system, controller, and its various units and modules can be found in the definitions of the cell sample processing method above and will not be repeated here. Each module in the controller can be implemented in whole or in part via software, hardware, or a combination thereof. As can be understood, as shown in FIG4 , the controller includes a processor, memory, a network interface, and a database connected via a system bus. Each module of the controller can be embedded in or independent of the processor in hardware form, or stored in the memory in software form, so that the processor can call and execute the corresponding operations of each module. The processor is used to provide computing and control capabilities. The memory includes a readable storage medium and an internal memory. The readable storage medium stores an operating system, computer-readable instructions, and a database. The internal memory provides an environment for the operation of the operating system and computer-readable instructions in the readable storage medium. The database is used to store data used in the cell sample processing method described in the above embodiments. The network interface is used to communicate with an external terminal via a network connection. The computer-readable instructions, when executed by the processor, implement a cell sample processing method. The readable storage medium provided in this embodiment includes a non-volatile readable storage medium and a volatile readable storage medium.

[0131] In one embodiment, one or more readable storage media storing computer-readable instructions are provided. The readable storage media provided in this embodiment include non-volatile readable storage media and volatile readable storage media. The readable storage media store computer-readable instructions. When the computer-readable instructions are executed by one or more processors, the one or more processors implement the above-mentioned cell sample processing method.

[0132] Those skilled in the art will understand that all or part of the processes in the above-mentioned embodiment methods can be implemented by instructing related hardware through computer-readable instructions. The computer-readable instructions can be stored in a non-volatile readable storage medium or a volatile readable storage medium. When the computer-readable instructions are executed, they can include the processes of the embodiments of the above-mentioned methods. Among them, any reference to memory, storage, database or other media used in the embodiments provided in this application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM) or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), Synchronous Link DRAM (SLDRAM), Rambus direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and memory bus dynamic RAM (RDRAM), etc.

[0133] Those skilled in the art will clearly understand that for the sake of convenience and brevity of description, only the division of the above-mentioned functional units and modules is used as an example. In actual applications, the above-mentioned functions can be distributed and completed by different functional units and modules as needed, that is, the internal structure of the device can be divided into different functional units or modules to complete all or part of the functions described above.

[0134] The above-described embodiments are only used to illustrate the technical solutions of the present application, rather than to limit them. Although the present application has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments can still be modified, or some of the technical features thereof can be replaced by equivalents. These modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present application, and should all be included in the scope of protection of the present application.

Claims

1. A method for processing a cell sample, wherein: include: The cell sample is flowed into a centrifuge container; Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid; Performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid; Activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid; genetically modifying the activated cell fluid to obtain target cell fluid; The target cell solution is divided into a plurality of preparation bags.

2. The cell sample processing method according to claim 1, wherein: The step of centrifuging the cell sample in the centrifugal container to obtain an initial cell solution comprises: Controlling the centrifugal container to rotate at a preset speed so that the cell sample in the centrifugal container can be centrifuged and layered; the cell sample includes at least one of T cells, T cell subsets and T cell progenitor cells; The initial cell fluid is separated from the cell sample after centrifugation and stratification.

3. The cell sample processing method according to claim 1, wherein: The method of subjecting the initial cell fluid to antigen binding and magnetic separation to separate the enriched cell fluid comprises: Adding magnetic beads and antigens to a centrifugal container containing the initial cell fluid, so that the magnetic beads and antigens are combined with the initial cells in the initial cell fluid to obtain a mixed cell fluid; The mixed cell fluid is subjected to magnetic bead sorting to sort out the enriched cell fluid.

4. The cell sample processing method according to claim 1, wherein: The step of activating the enriched cell fluid by inputting a regulator to obtain the activated cell fluid comprises: injecting the regulator into a centrifuge container containing the enriched cell fluid; The enriched cells in the enriched cell fluid are activated by the co-stimulator in the regulator to obtain the activated cell fluid; the regulator is one or more of agonistic antibodies, cytokines, recombinant co-stimulatory molecules and small drug inhibitors.

5. The cell sample processing method according to claim 4, wherein: The step of activating the enriched cells in the enriched cell fluid by the co-stimulator in the regulator to obtain the activated cell fluid comprises: The co-stimulator in the regulator causes the enriched cells in the enriched cell fluid to produce a co-stimulatory effect, thereby obtaining active cell fluid; Sampling and testing the active cell fluid to obtain a test result; When the detection result indicates that the enriched cells have been successfully activated, the active cell fluid is determined as the activated cell fluid.

6. The cell sample processing method according to claim 5, wherein: The co-stimulator comprises at least one of an anti-CD3 antibody or fragment and an anti-CD28 antibody or fragment; or / and The co-stimulator includes a nanomatrix including at least one of a matrix of mobile polymer chains and a matrix attached to the mobile polymer chains.

7. The cell sample processing method according to claim 1, wherein: The step of genetically modifying the activated cell fluid to obtain the target cell fluid comprises: performing transduction treatment on the activated cell fluid to obtain the target cell fluid; or performing transfection treatment on the activated cell fluid to obtain the target cell fluid; or Performing electroporation on the activated cell fluid to obtain the target cell fluid; or The activated cell fluid is subjected to mechanical transfection treatment to obtain the target cell fluid.

8. The cell sample processing method according to claim 7, wherein: The step of performing transduction treatment on the activated cell fluid to obtain the target cell fluid comprises: The activated cell fluid is transduced by a transduction enhancer to obtain the target cell fluid; the transduction enhancer includes one or more of a polycationic agent, poloxamer, poloxamer, and an adhesion molecule.

9. A cell sample processing device, wherein: include: A sample inflow module, used for flowing the cell sample into a centrifugal container; A cell separation module, used for centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid; A cell enrichment module, used for performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid; A cell activation module, used to activate the enriched cell fluid by inputting a regulator to obtain an activated cell fluid; A genetic modification module, used for genetically modifying the activated cell fluid to obtain a target cell fluid; The preparation packaging module is used to package the target cell solution into multiple preparation bags.

10. The cell sample processing device according to claim 9, wherein: The step of centrifuging the cell sample in the centrifugal container to obtain an initial cell solution comprises: Controlling the centrifugal container to rotate at a preset speed so that the cell sample in the centrifugal container can be centrifuged and layered; the cell sample includes at least one of T cells, T cell subsets and T cell progenitor cells; The initial cell fluid is separated from the cell sample after centrifugation and stratification.

11. The cell sample processing device according to claim 9, wherein: The method of subjecting the initial cell fluid to antigen binding and magnetic separation to separate the enriched cell fluid comprises: Adding magnetic beads and antigens to a centrifugal container containing the initial cell fluid, so that the magnetic beads and antigens are combined with the initial cells in the initial cell fluid to obtain a mixed cell fluid; The mixed cell fluid is subjected to magnetic bead sorting to sort out the enriched cell fluid.

12. The cell sample processing device according to claim 9, wherein: The step of activating the enriched cell fluid by inputting a regulator to obtain the activated cell fluid comprises: injecting the regulator into a centrifuge container containing the enriched cell fluid; Activating the enriched cells in the enriched cell fluid by the co-stimulator in the regulator The activated cell fluid is obtained by treatment; the regulator is one or more of agonistic antibodies, cytokines, recombinant co-stimulatory molecules and small drug inhibitors.

13. The cell sample processing device according to claim 12, wherein: The step of activating the enriched cells in the enriched cell fluid by the co-stimulator in the regulator to obtain the activated cell fluid comprises: The co-stimulator in the regulator causes the enriched cells in the enriched cell fluid to produce a co-stimulatory effect, thereby obtaining active cell fluid; Sampling and testing the active cell fluid to obtain a test result; When the detection result indicates that the enriched cells have been successfully activated, the active cell fluid is determined as the activated cell fluid.

14. The cell sample processing device according to claim 13, wherein: The co-stimulator comprises at least one of an anti-CD3 antibody or fragment and an anti-CD28 antibody or fragment; or / and The co-stimulator includes a nanomatrix including at least one of a matrix of mobile polymer chains and a matrix attached to the mobile polymer chains.

15. The cell sample processing device according to claim 9, wherein: The step of genetically modifying the activated cell fluid to obtain the target cell fluid comprises: performing transduction treatment on the activated cell fluid to obtain the target cell fluid; or performing transfection treatment on the activated cell fluid to obtain the target cell fluid; or Performing electroporation on the activated cell fluid to obtain the target cell fluid; or The activated cell fluid is subjected to mechanical transfection treatment to obtain the target cell fluid.

16. The cell sample processing device according to claim 15, wherein: The step of performing transduction treatment on the activated cell fluid to obtain the target cell fluid comprises: The activated cell fluid is transduced by a transduction enhancer to obtain the target cell fluid; the transduction enhancer includes one or more of a polycationic agent, poloxamer, poloxamer, and an adhesion molecule.

17. A cell sample processing method system, wherein: The invention comprises a centrifugal container and a controller connected to the centrifugal container, wherein the controller is used to perform the following cell sample processing method: The cell sample is flowed into a centrifuge container; Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid; Performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid; Activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid; genetically modifying the activated cell fluid to obtain target cell fluid; The target cell solution is divided into a plurality of preparation bags.

18. One or more computer-readable storage media storing computer-readable instructions, wherein: When the computer readable instructions are executed by one or more processors, the one or more processors perform the following steps: The cell sample is flowed into a centrifuge container; Centrifuging the cell sample in the centrifugal container to obtain an initial cell fluid; Performing antigen binding and magnetic separation on the initial cell fluid to separate the enriched cell fluid; Activating the enriched cell fluid by inputting a regulator to obtain an activated cell fluid; genetically modifying the activated cell fluid to obtain target cell fluid; The target cell solution is divided into a plurality of preparation bags.

19. The computer-readable storage medium of claim 17, wherein: The step of activating the enriched cell fluid by inputting a regulator to obtain the activated cell fluid comprises: injecting the regulator into a centrifuge container containing the enriched cell fluid; The enriched cells in the enriched cell fluid are activated by the co-stimulator in the regulator to obtain the activated cell fluid; the regulator is one or more of agonistic antibodies, cytokines, recombinant co-stimulatory molecules and small drug inhibitors.

20. The computer-readable storage medium of claim 19, wherein: The step of activating the enriched cells in the enriched cell fluid by the co-stimulator in the regulator to obtain the activated cell fluid comprises: The co-stimulator in the regulator causes the enriched cells in the enriched cell fluid to produce a co-stimulatory effect, thereby obtaining active cell fluid; Sampling and testing the active cell fluid to obtain a test result; When the detection result indicates that the enriched cells have been successfully activated, the active cell fluid is determined as the activated cell fluid.

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