70 results about "Single cell suspension" patented technology

The embodiment of the invention relates to the technical field of drug detection, in particular to an anti-CD38 antibody drug receptor occupation detection method and system, a peripheral blood sample is used for preparing a single-cell suspension, the single-cell suspension is divided into a TEST tube and an FMO tube, a cell surface antibody and an RO detection antibody are added into the TEST tube, only the cell surface antibody is added into the FMO tube, and only the RO detection antibody is added into the FMO tube. The TEST tube and the FMO tube are subjected to sample detection analysis, so that the average fluorescence intensity of RO detection antibodies of the TEST tube and the FMO tube is obtained. And performing RO detection analysis by adopting a dissociation method, taking the tubes which are not combined with the medicine as a total receptor group, taking the tubes which are combined with the medicine as a dissociation receptor group, and calculating an RO detection result according to the average fluorescence intensity of the TEST tube and the FMO tube.

The present disclosure relates to a method of generating pancreatic lineage cells, e.g. islet-like cell aggregates comprising pancreatic beta cells, comprising the steps of providing a single cell suspension of a population of endocrine progenitor cells (EP), allowing the EP cells in the single cell suspension to form 3D structures, and culturing the cells under conditions allowing differentiation into pancreatic mono-hormonal beta cells. The present disclosure also relates to islet-like cell aggregates obtainable by said method and their medical use.

This invention relates to a method for detecting colorectal cancer tumor cell-associated microorganisms, specifically a method for eliminating intestinal fecal microbial contamination and detecting colorectal cancer tumor-associated microorganisms, belonging to the field of biotechnology. This invention initially eliminates fecal microbial contamination from the sample structure by coarsely shearing and enzymatically digesting the intestinal folds and villi; then, it uses a 40μm cell sieve to filter out incompletely digested tissue fragments, further eliminating intestinal fecal bacteria contamination, and collecting a single-cell suspension; subsequently, the collected single-cell suspension is repeatedly washed with 300g centrifugation to remove as many intercellular or loosely bound "transient bacteria" as possible, retaining intracellular and firmly colonized microorganisms. Ultimately, this eliminates the severe contamination from the large and complex fecal microorganisms, allowing the detection of tumor cell-associated microorganisms, which are already present in low biomass. This method is suitable for detecting microorganisms in gastrointestinal tissues that are in direct contact with the environment.

The invention belongs to the cross technical field of veterinary medicine and biomedicine, and relates to a construction and evaluation method of a dog spontaneous breast cancer organoid model, which specifically comprises the following steps: (1) obtaining dog spontaneous breast cancer tissues through operation or biopsy, separating tumor tissues under sterile conditions, and carrying out fine enzymolysis digestion treatment to obtain dog spontaneous breast cancer tissues; obtaining a single-cell suspension; and (2) inoculating the single-cell suspension into a three-dimensional culture system, carrying out three-dimensional culture by adopting a culture medium containing EGF, FGF and B27 in combination with a growth factor, promoting the growth and amplification of the breast cancer organs, and carrying out timely passage according to the growth condition so as to maintain the activity of the model. The model constructed by the invention completely retains genetic heterogeneity and pathological microenvironment of primary tumors, does not need gene editing, is simple and convenient to operate and low in cost, is particularly suitable for individual drug sensitivity detection of pet dogs, and provides an efficient and high-simulation experimental platform for dog breast cancer mechanism research and clinical precise medication.

The invention discloses a non-tumorigenic MDCK cell strain, a screening method and application. The non-tumorigenic MDCK cell strain is MDCK-CA027 (the preservation number is CCTCC (China Center for Type Culture Collection) NO: C2023329). The tumor formation rate of the cell strain is 0% when the cell strain is observed for at least 16 weeks in a nude mouse in-vivo tumor formation test, and the cell strain has good sensitivity to various influenza viruses (such as H1N1, H3N2, BY and BV). The invention also provides a method for screening the cell strain. The method comprises the following steps: preparing a single-cell suspension, carrying out limited dilution to obtain monoclone, carrying out enlarged culture, carrying out virus sensitivity detection, carrying out in-vitro tumorigenicity related detection (clone formation test / soft agar test), carrying out nude mouse in-vivo tumorigenicity test and the like. The non-tumorigenic MDCK cell strain is high in safety, can efficiently support influenza virus replication, can be safely and effectively applied to influenza vaccine production and influenza virus amplification, overcomes the tumorigenic risk of traditional MDCK cells, and has important scientific significance and application prospects.

This invention provides a method and apparatus for splitting single-cell mixed sample sequencing data, relating to the field of biotechnology. The splitting method includes: capturing and sequencing single-cell suspensions using a single-cell platform; then performing reference genome alignment, cell identification, and gene expression level quantification on the sequencing data using Cellranger; splitting the cell data identified in step a into two groups of cell data for different sexes based on SNP locus information from the 1000 Genomes Project; and distinguishing the two groups of cell data from male or female samples based on the proportion of sex-specific genes expressed in the two groups of cell data. This splitting method eliminates the need for additional experimental operations such as protein labeling and genome sequencing, and can provide accurate and reliable data splitting even when individual SNP information is unavailable.

The invention belongs to the field of immunotoxicology, and particularly relates to a method for immunotyping bone marrow cells by flow cytometry. The flow cytometry immune typing method for bone marrow cells comprises the following steps: collecting macaca fascicularis bone marrow fluid, and preparing the macaca fascicularis bone marrow fluid into single-cell suspension; dyeing the obtained cells by using an antibody combination containing a plurality of fluorescent labels; then, a flow cytometer is used for collecting cell fluorescence signals, a standardized gate setting strategy is executed, and based on collected data, the percentage of each cell subset in CD45 + living white blood cells is calculated to complete immune typing. According to the flow cytometry immune typing method for the bone marrow cells, the problems of low efficiency, low repeatability, low sensitivity and poor specificity of related technologies are solved, and more accurate and reliable data support is provided for early discovery, risk assessment and mechanism research of drug immunotoxicity.

The application discloses a kind of mesenchymal stem cell ball fractional dissociation digestive juice, its preparation method and application, it is related to cell preparation and three-dimensional cell culture technical field, specifically at to provide C liquid, D liquid and E liquid three kinds of gradient concentration working solution.When dissociating, the mesenchymal stem cell ball to be dissociated is determined its average particle size;According to the average particle size, select C liquid, D liquid or E liquid as digestive juice as described above;The selected digestive juice is mixed with the mesenchymal stem cell ball, and digestion reaction is carried out under oscillation condition, and single cell suspension is obtained.The application establishes the hierarchical matching rule of mesenchymal stem cell ball particle size and specific digestive juice concentration, and optimizes oscillation digestion condition, so as to maintain high activity rate and complete phenotype characteristics while efficiently dissociating, realizes the standardization of process and the controllable of dissociation quality, provides stable and reliable solution for the large-scale preparation and quality control of mesenchymal stem cell ball.

The invention discloses a cell crude sample sex determination primer suitable for chickens, ducks or pigeons and a PCR detection method, and belongs to the technical field of biological detection. A pair of gender-specific universal primers for various poultry such as chicken, duck and pigeon is designed for the first time, a cell crude sample is adopted as a PCR template, stable amplification can be realized without DNA extraction, the operation process is greatly simplified, the detection time is shortened, and the detection cost is reduced. Meanwhile, TrypLE enzyme digestion is very mild to cells, and the method is also suitable for preparation of single-cell suspensions related to single-cell omics. The method is suitable for the fields of early sex identification of poultry, breeding poultry breeding, molecular breeding and the like, and has wide application prospects.

The invention belongs to the technical field of biology, and particularly relates to a method for quantitatively detecting the infection rate of lentivirus on target cells. Comprising the following steps: infecting target cells with a lentivirus-containing sample to be detected; preparing a single-cell suspension from the target cells integrated with the lentivirus, uniformly dividing the single-cell suspension into two parallel samples which are identical in number and state, and inoculating the parallel samples to obtain antibiotic treated cells and antibiotic-free treated cells; adding antibiotics into the medicine sieve group for screening, counting to obtain positive infected cells Ne, and counting the control group without adding the medicines at three time points, namely when the dichotomy operation is executed, when the antibiotics start to screen and after the antibiotics are screened, so as to obtain the total cell numbers No.2, No.1 and No.3; and calculating the infection rate result of the lentivirus on the target cells based on the total cell number. According to the method, the sampling time node of the control group is optimized, the cell number of the control group is selected as the denominator of the infection rate calculation when the medicine sieve group samples start to be added, and the accuracy of the experimental result is further improved.

The invention discloses a single-cell suspension purification method capable of maintaining representativeness of a cell population and a kit, and belongs to the technical field of single-cell separation. The method comprises the following steps that a cell sample to be separated and purified is resuspended in a centrifugal container through a reagent B, then a reagent A is added and evenly mixed, the adding amount of the reagent A and the reagent B is adjusted so that the density of the suspension can be 1.08-1.15 g / ml, the reagent A is an aqueous solution containing iodixanol, and the reagent B is a DMEM culture medium containing 1-10% of FBS; and slowly laying PBS on the upper layer of the suspension, centrifuging, collecting living cells of an interface layer, washing the obtained living cells, and re-suspending. According to the scheme, the single-cell suspension for single-cell library construction can be purified under the conditions of low equipment requirement, low cost and mild conditions, the cell population representativeness of a sample can be reserved to the maximum extent while dead cells are removed, and no adverse effect is generated on the single-cell library construction process.

The application provides a solid tumor targeting index detection method and a reagent combination thereof, including detection of a solid tumor targeting therapy molecular marker and detection of a solid tumor microenvironment, and the evaluation method comprises the following steps: collecting a fresh tumor tissue sample; preparing a tumor tissue sample to be detected into a single cell suspension; establishing a flow cytometry on-machine model; detecting the solid tumor targeting therapy molecular marker by using flow cytometry; and detecting the solid tumor microenvironment by using flow cytometry. The application has the advantages of less required samples, short detection time, and the ability to provide information about various factors that are helpful for prognosis and treatment, including quantification of tumor infiltrating lymphocytes, the proportion of tumor cells with positive expression of PD-L1, EGFR and ALK, and the proportion of tumor cells with co-expression.

The invention relates to the technical field of molecular immunology and biology, in particular to a sicca syndrome model mouse B cell immune repertoire sequencing and subpopulation analysis method. The method comprises the following steps: S1, sample preparation: preparing a single-cell suspension from a target autoimmune disease animal model and a preset tissue of a control animal, and carrying out B cell enrichment treatment on the single-cell suspension to obtain a B cell sample; s2, multi-dimensional data parallel analysis: analyzing the B cell sample, and performing parallel construction of the multi-dimensional data; and S3, integrated correlation analysis: carrying out correlation analysis on the immune group library data, the cell subset data and the signal channel data obtained in the step S2 to establish a correlation relationship among specific B cell cloning characteristics, specific B cell subset changes and specific signal channel activity, and carrying out multilevel analysis and generating a B cell immune response mechanism of the autoimmune disease. The efficiency and accuracy of disease mechanism research and drug action mechanism evaluation are remarkably improved.

The invention relates to the technical field of biomedicine, in particular to a method for identifying fibroblast subsets in colorectal cancer tissue based on single cell sequencing, and provides a method for identifying fibroblast subsets in colorectal cancer tissue based on single cell sequencing. The method comprises the following steps: S1, extracting colorectal cancer tissues: obtaining tumor tissues removed from the colorectal cancer tissues, and collecting a suspension to prepare a single-cell suspension; s2, single cell sequencing analysis: constructing a high-throughput sequencing library; s3, data analysis: processing the sequencing data by using a CellRanger software pipeline, and mapping readings to a genome and a transcriptome; and carrying out data dimension reduction and visualization. S4, identification of the tumor-associated fibroblast subpopulation: after the total cell type in the colorectal cancer tumor tissue is identified, annotating the cells; and identifying differentially expressed genes by using a Seurt software package, and carrying out function enrichment analysis, so as to identify various cell types.

The invention belongs to the technical field of single-cell suspension preparation, and discloses a single-cell suspension preparation device convenient to take, the single-cell suspension preparation device comprises a rotating shaft and blades distributed on the surface of the rotating shaft, the rotating shaft is connected with a power source outputting rotation action, the single-cell suspension preparation device further comprises a cylinder and a cleaning mechanism, and the rotating shaft is installed in the cylinder and only has rotation freedom; one end of the barrel is detachably connected with a filter cover provided with a liquid outlet, the filter cover is provided with a filter basket, the filter basket intercepts solid substances moving towards the liquid outlet, and the liquid outlet is provided with a valve; the cleaning mechanism comprises a liquid return pipeline detachably connected with the valve, the liquid return pipeline is connected with a water pump and a first flow adjusting valve in series, the end, opposite to the end connected with the valve, of the liquid return pipeline is detachably connected with a spray head, and the spray head is installed on the cylinder wall of the cylinder and stretches into the cylinder. The liquid return pipeline is connected with a liquid supply pipeline beside the line position between the water pump and the first flow adjusting valve, the end of the liquid supply pipeline is connected with a cleaning liquid supply end, and a second flow adjusting valve is arranged on the line of the liquid supply pipeline.

The application discloses a single-cell immunoblotting method based on quenchable aptamer probes and relates to the technical field of single-cell protein detection. The method comprises the following steps: S1, a single-cell suspension sample is settled in a microwell array on a composite hydrogel chip; S2, the composite hydrogel chip is placed in an electrophoresis tank, then a lysis solution is slowly poured into the tank, and incubation lysis is carried out; S3, after cell lysis, an electric field is applied to both ends of the chip, and proteins enter a gel coating layer on the surface of the chip under the action of the electric field and start electrophoretic separation; S4, after gel electrophoresis is completed, the chip is placed under an ultraviolet lamp for exposure, and the gel is taken out after exposure is completed; S5, the gel is placed in an aptamer probe solution which is targeted to target proteins, and is soaked and dyed, and the fluorescence signal intensity of the target protein molecules is measured under a laser confocal fluorescence microscope. The application can improve the detection sensitivity of target proteins and has fast imaging speed.

The invention discloses a preparation method and application of a prostate cancer tissue single-cell suspension. The preparation method of the prostate cancer tissue single-cell suspension comprises the following steps: S1, tissue pretreatment: putting a prostate cancer tissue sample into a low-temperature pretreatment buffer solution, and cutting the prostate cancer tissue sample into tissue fragments; s2, circular enzymolysis: mixing the tissue fragments with a composite enzymatic hydrolysate, and performing digestion for multiple rounds under an oscillation condition; after each round of digestion, collecting supernate into a stop solution containing serum, repeatedly digesting the remaining tissues for multiple times, and combining the supernate to obtain a cell crude suspension; wherein the composite enzymatic hydrolysate contains collagenase, protease, nuclease, an androgen receptor stimulant and / or a ROCK inhibitor; and S3, sequentially carrying out separation and purification on the cell crude suspension through filtration and density gradient centrifugation to obtain the prostate cancer tissue single-cell suspension.

The application provides a preparation method and application of a retinal nerve vascular unit in-vitro 3D printing model, and relates to the field of biomedical technology, and comprises the following steps: preparing a composite hydrogel precursor solution by using GelMA, HAMA and LAP; selecting RECs, RGCs and RMCs in an exponential growth phase, respectively resuspending after trypsin digestion, mixing the three single-cell suspensions to obtain a cell mixture; mixing the composite hydrogel precursor solution and the cell mixture to obtain a biological ink; printing and packaging the biological ink according to preset parameters to form a preliminary 3D-RNVU model; and culturing the preliminary 3D-RNVU model again. The 3D-RNVU model constructed in the application realizes the reproduction of the physical and chemical properties of ECM and the neural-glial-vascular three-way interaction in a three-dimensional structure, and provides an in-vitro platform with high physiological correlation for mechanism research and drug screening of diseases such as diabetic retinopathy.

The embodiment of the invention relates to a method for controlling a tissue dissociation system and the tissue dissociation system. The method comprises the following steps: at a control device, controlling a tissue dissociation instrument to dissociate a tissue sample in a dissociation tube, and providing the dissociated tissue sample to a liquid workstation through a mechanical arm; treating the dissociated tissue sample by a liquid workstation, and sub-packaging the obtained cell sample into a cell counting sample plate; a mechanical arm provides the cell counting sample plate to a cell culture box, and the cell culture box performs incubation operation on cells in the cell counting sample plate; the cell counting sample plate incubated by the cell incubator is transferred to an image analysis system by the automatic guided vehicle; the image analysis system is used for carrying out cell counting on the cell counting sample plate and sending a cell counting result to the liquid workstation; and based on the cell counting result and the predetermined density value, controlling the liquid workstation to prepare the single-cell suspension with the predetermined density. In this way, tissue dissociation automation is achieved.

The invention discloses a method and a kit for rapidly screening gene editing positive monoclone based on qPCR (quantitative polymerase chain reaction), and belongs to the field of gene engineering. The method comprises the following steps: preparing a monoclonal sample: preparing a single-cell suspension from cells subjected to gene editing, and carrying out single-cell sorting and culture until monoclone is formed; collecting monoclonal cells, and directly cracking by using a cracking solution to prepare a qPCR template; qPCR detection: carrying out qPCR amplification and detection on the sample by using a specific primer and a probe designed aiming at a target gene editing event; and result judgment: judging whether the monoclone is gene editing positive clone or not by analyzing the Ct value of the qPCR. According to the present invention, the specific primers and the probe of the target gene editing site are designed, and the qPCR is combined to directly detect the genome variation of the monoclonal cell so as to achieve the early rapid screening;

The invention discloses a reagent composition capable of being used for flow cytometry detection of erythrocyte line cell differentiation and application of the reagent composition. The reagent composition disclosed by the invention is used for being added into a flow type tube in a single cell suspension state of a sample to be detected, and comprises an anti-CD71 antibody, an anti-CD117 antibody, an anti-CD34 antibody, an anti-CD105 antibody, an anti-CD36 antibody and an anti-CD45 antibody. The reagent composition disclosed by the invention can be applied to flow cytometry for detecting the cell differentiation condition of the red blood cell line, screening and / or diagnosing and follow-up visit, so that high-sensitivity and specific diagnosis is realized, follow-up visit after treatment can be realized, a standardized scheme is formed, the efficiency and the accuracy are improved, and convenience is provided for realizing automation and artificial intelligence in the future.

This invention discloses a method for assessing the sensitivity of anticancer drugs based on patient-derived tumor tissue, belonging to the field of biomedical technology. The method includes: obtaining fresh tumor tissue from a patient and preparing it into tissue fragments of approximately 1 mm³; culturing the tissue fragments in vitro in a three-dimensional gel matrix; adding the anticancer drug to be tested to the culture system; digesting the tissue fragments after culture to obtain a single-cell suspension; and assessing drug sensitivity by detecting cell viability using flow cytometry. This invention can maximally preserve tumor heterogeneity and microenvironment, has a short operation cycle, and can quickly and accurately guide personalized clinical medication, especially suitable for solid tumors such as brain tumors.

The invention relates to the technical field of biomedical detection, in particular to a multiple myeloma circulating plasma cell flow cytometry method and system, a peripheral blood sample is used for preparing a single-cell suspension, the single-cell suspension is extracted to prepare a to-be-detected tube (TEST tube), the TEST tube is subjected to non-specific site sealing, and the TEST tube is subjected to non-specific site sealing to obtain the multiple myeloma circulating plasma cell flow cytometry method and the multiple myeloma circulating plasma cell flow cytometry system. The method comprises the following steps: taking a TEST tube as a substrate, adding a multi-fluorescent antibody panel containing nano antibodies aiming at BCMA and GPRC5D to perform cell staining operation, performing flow cytometry analysis on the TEST tube to obtain flow cytometry data, and performing gate analysis on the flow cytometry data according to a preset detection analysis strategy to obtain a fluorescence detection result. The number and phenotypic characteristics of circulating plasma cell (CPC) populations are identified and counted, so that circulating plasma cell detection information is generated.

This invention provides a method to improve age-related decline in ovarian function by supplementing quiescent oocytes within primordial follicles. [Solution] The present invention provides a method for inducing mature oocytes, comprising the steps of: preparing a single-cell suspension from the ovary of a mammal other than a human; separating the single-cell suspension into oocytes and ovarian somatic cells (supporting cells) within a primordial follicle using a flow cytometer; re-aggregating the oocytes and ovarian somatic cells within the separated primordial follicle; and maturing the re-aggregated primordial follicle in vitro.

This invention discloses a method for the isolation, culture, and identification of chicken sexual gonadal cells, belonging to the field of biotechnology. The invention selects chicken embryos hatched to 18.5 days old, isolates the gonads after PCR sex identification, and uses 0.25% trypsin-EDTA digestion followed by filtration through a 70μm filter to obtain a single-cell suspension with a survival rate >90% and free from PGC contamination. A sex-specific culture system is established, allowing cells to be stably passaged and maintain a sex-specific phenotype. Through morphological, qRT-PCR, immunofluorescence, and Western blot multidimensional identification, accurate identification of male testicular Sertoli cells and female ovarian granulosa cells is achieved. This method is simple to operate, has good reproducibility, and produces high cell viability and purity. It can provide a stable gonadal microenvironment for the in vitro directed differentiation of chicken primordial germ cells, significantly improving gamete induction efficiency. It is suitable for research on avian reproductive and developmental mechanisms, transgenic breeding, and germplasm resource preservation.

The invention discloses a preparation method and application of a biological agent for promoting tendon aging repair, relates to the field of treatment of tendon injury caused by aging, and aims to solve the problems of insufficient cell regeneration capacity and increase of tendon injury caused by tendon aging. The method comprises the following steps: 1, immersing activated graphene oxide aerogel into a fisetin solution, standing, and fixing to obtain fisetin-loaded graphene oxide aerogel; 2, putting the fisetin-loaded graphene oxide aerogel into a culture device, adding the single-cell suspension into the culture device, and carrying out cell culture; and 3, collecting cell supernatant, performing low-intensity pulse ultrasonic treatment, performing ultrafiltration concentration, and adding normal saline to prepare the biological preparation for promoting tendon aging repair. The biological preparation prepared by the invention can slow down tendon aging, improve tendon injury and promote tendon generation.

1. The name of the design product: electronic device with single cell suspension preparation instrument operation program graphical user interface. 2. The use of the design product: an electronic device for single cell suspension preparation instrument. 3. The design points of the design product: the graphical user interface of the single cell suspension preparation instrument operation program. The hardware design part and the rest are designed as usual. 4. The picture or photo that best indicates the design points: interface change state diagram 2. 5. The use of the graphical user interface: the interactive interface of the single cell suspension preparation instrument during operation. 6. The change state of the graphical user interface: the main view shows the start-up interface; interface change state diagram 1 is the graphical user interface after the start-up is completed; interface change state diagram 2 is the graphical user interface after clicking the "Protocols" option on the left side of interface change state diagram 1; interface change state diagram 3 is the graphical user interface after clicking the "New" option in the upper right corner of interface change state diagram 2; interface change state diagram 4 is the graphical user interface after clicking the left option among the three options in the upper right corner of interface change state diagram 3.

The invention provides a preparation method and application of a retinal nerve and blood vessel unit in-vitro 3D printing model, and relates to the technical field of biomedicines.The preparation method comprises the steps that GelMA, HAMA and LAP are used for preparing a composite hydrogel precursor solution; the method comprises the following steps: selecting REC, RGC and RMC in an exponential growth phase, digesting the REC, RGC and RMC with trypsin respectively, resuspending, and mixing three single-cell suspensions to obtain a cell mixture; mixing the composite hydrogel precursor solution with the cell mixture to obtain biological ink; the bio-ink is printed and packaged according to preset parameters, and a preliminary 3D-RNVU model is formed; and the preliminary 3D-RNVU model is cultured again. According to the constructed 3D-RNVU model, the physical and chemical characteristics of ECM and nerve-colloid-blood vessel ternary interaction are represented in a three-dimensional structure, and an in-vitro platform with high physiological correlation is provided for mechanism research and drug screening of diseases such as diabetic retinopathy.

The invention provides application of polyethylene glycol in reducing diffusion of cell contents in a single-cell suspension and improving single-cell capture efficiency. When the single-cell suspension containing polyethylene glycol is used for capturing single-cell nucleic acid, diffusion of contents of single cells, such as nucleic acid such as genome DNA or transcriptome RNA, can be effectively reduced, so that the contents of the single cells are more gathered, and the cell flux and capturing efficiency are effectively improved.

The invention discloses a construction and evaluation method of an Rb1 and Trp53 (p53) double-gene mutation mouse spontaneous lung cancer organoid model, and the construction method comprises the following steps: separating lung cancer tissues under a sterile condition, and carrying out fine enzymolysis digestion treatment to obtain a single-cell suspension; inoculating the single-cell suspension into a three-dimensional culture system, carrying out three-dimensional culture by adopting a culture medium containing EGF, FGF and B27 in combination with growth factors, promoting the growth and amplification of lung cancer organs, and carrying out timely passage according to the growth condition to maintain the activity of the model; the model constructed by the invention completely retains genetic heterogeneity and pathological microenvironment of the primary tumor, in-vitro gene editing is not needed, the operation is simple and convenient, the cost is low, and an efficient and high-simulation experimental platform is provided for lung cancer mechanism research and individualized drug screening.