A device for a precise tumor treatment method based on CTC circulating tumor cells

Abstract

The present invention provides a device that can be used for a precise tumor treatment method based on CTC circulating tumor cells. It includes the following steps: a CTC circulating tumor cell separation device, which is used to separate CTC circulating tumor cells from the added peripheral blood; a CTC circulating tumor cell detection device, which is used to detect the separated CTC circulating tumor cells; a closable CTC circulating tumor cell culture device, which is used to culture and proliferate the separated CTC circulating tumor cells in vitro; a drug sensitivity detection device, which is used to perform drug sensitivity experiments on the cultured CTC circulating tumor cells. The present invention uses the patient's CTC circulating tumor cells for in vitro drug sensitivity testing, which has a personalized and precise treatment effect for the patient; the method of the present invention for parallel drug sensitivity testing can optimize a clinical medication regimen each time, so that tumor cells are always facing the attack of different types of drugs, thereby weakening the ability of the tumor to adapt to the drug, and avoiding the drug resistance problem caused by a single clinical drug.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to a device that can be used for a precise tumor treatment method based on CTC circulating tumor cells. Background Art

[0002] Cancer is a major disease in today's world. Its high mortality rate and complexity of treatment have prompted countries around the world to spend a lot of manpower and material resources on research, but so far no breakthrough has been made. It is called the king of all diseases by American physician Siddhartha Mukherjee.

[0003] The formation of tumors is a complex process involving multiple factors, which makes its research more complicated. The treatment of tumors is based on the understanding of tumors, and has gone through surgery, radiotherapy, conventional chemotherapy, targeted drug therapy, and the current hot research method, immune cell therapy.

[0004] Surgery, as the name implies, is the removal of tumors visible to the naked eye. The difficulty of this method is that the relationship between resection and tumor is uncertain. Due to the complexity of tumors, especially in the middle and late stages, distant tissues of the patient may have metastasized and infiltrated, so surgery can only remove part of it and it is difficult to remove it completely. Take brain tumors as an example. Because it is difficult to distinguish and remove tumors smaller than 1mm with the naked eye, and highly malignant tumor cells such as gliomas have already spread in normal brain tissues, the tumor will recur soon after surgery. Secondly, even if the tumor is removed cleanly, if the tumor is not the primary tumor, then primary tumors in other parts of the body will develop. Therefore, surgery is currently only recommended as a method for the treatment of early tumors.

[0005] Radiotherapy is the use of high-power rays to destroy tumor tissue. The difficulty of this method is how to accurately capture tumor tissue and perform high-precision radiotherapy on it. Even if the range is very accurate, due to the three-dimensional structure of human tissue, the problem of normal tissue destruction in the depth direction will occur when the human body is irradiated with rays in a straight line. The minimum degree of damage is to not damage the normal human tissue behind the tumor tissue. Therefore, radiotherapy is currently only used as a means to alleviate the symptoms of patients and cannot fundamentally prevent the development of tumors.

[0006] Conventional chemotherapy, because it is also toxic to normal cells, will cause serious side effects to patients, causing a significant decline in their physical condition, and making them unfit for further chemotherapy. This method is a phased treatment measure and cannot be used for sustained treatment.

[0007] Targeted drug therapy mainly intervenes in some specific gene mutations in tumor cells to achieve the purpose of inhibiting and killing tumor cells. The advantage of targeted drugs is that they greatly reduce the toxic side effects of conventional chemotherapy, but there are two difficulties. One is that the tumor cells of many cancer patients do not have gene mutations that can be intervened by targeted drugs. Currently, only 5% of patients can be treated with targeted drugs. Even for this part of patients, another difficulty in targeted drug treatment is drug resistance. Due to the complexity of the pathogenesis of tumors, one or several targets cannot control the tumor at all. Therefore, targeted drugs can only delay the progression of tumors, but cannot fundamentally control tumors.

[0008] Tumor immunotherapy is the application of immunological principles and methods to improve the immunogenicity of tumor cells and their sensitivity to effector cell killing, stimulate and enhance the body's anti-tumor immune response, and infuse immune cells and effector molecules into the host to cooperate with the body's immune system to kill tumors and inhibit tumor growth. At present, some tumor immunotherapy drugs have been approved by the US FDA and have entered clinical application. Existing immunotherapy includes checkpoint therapy and ACT (adoptive cell transfer) cell transfusion therapy. In 2011, the first approved immune checkpoint drug was Bristol-Myers Squibb's CTLA4 inhibitory monoclonal antibody lpilimumab (Yervoy). When doctors and medical scientists have new anti-tumor drugs, it is natural to think of the issue of combined medication. With the emergence of the second-generation immune checkpoint inhibitors, which target PD-1 / PD-L1, the FDA has accelerated the approval of a large number of immune checkpoint drugs, such as pembrolizumab for the treatment of malignant melanoma, nirolumab for the treatment of melanoma and lung cancer, and so on. The combined use of PD-1 and CTLA4 drugs has greatly improved the objective response rate of patients with malignant melanoma. Because the tumor microenvironment is significantly suppressive, it is a better choice to first contact the immunosuppressive environment with immune checkpoints.

[0009] Combination immunotherapy treatments include:

[0010] (1) Using immune checkpoints as the backbone of the basic selection. For example, using CTLA4 and PD-1 inhibitors at the same time, because although CTLA4 and PD1 are both expressed on T cells, their inhibitory mechanisms are different. CTLA4 competes with CD28 for the CD80 / 86 signaling pathway, thereby activating T cells, while PD-1 is expressed on activated lymphocytes.

[0011] (2) Combined with chemotherapy: Chemotherapy is used first to reduce the tumor burden, followed by immunotherapy.

[0012] (3) Combined with BRAF inhibitors.

[0013] (4) Combined with VEGF-targeted drugs.

[0014] In short, the goal of immunotherapy is to achieve a long-term stable therapeutic effect. However, due to the complexity of the tumor formation mechanism, the above applications are still in clinical research, and their effectiveness on patients remains to be observed.

[0015] In summary, due to the complexity of tumors, traditional treatment methods can only play a very limited role. So far, tumors are a disease whose pathogenesis is not clear, treatment methods are extremely limited, and the course of the disease is changing.

[0016] Different tumors have different characteristics due to different primary sites; the same tumor can be very different between different patients; in the same patient, the phenotype and gene mutation of tumor cells are different in primary lesions and metastatic lesions, or even in different locations of the same tumor tissue. Therefore, in the field of tumor treatment, there is a lack of a precise treatment method that integrates various sub-fields and takes into account the systemic considerations of patients, treatments, and research. Summary of the invention

[0017] In view of the problem that drug resistance is easily generated during the current tumor treatment process, the present invention provides a device that can be used for a precise tumor treatment method based on CTC circulating tumor cells.

[0018] The first aspect of the present invention is to provide a device for a precise tumor treatment method based on CTC circulating tumor cells, comprising:

[0019] - a housing, wherein the housing is provided with a hole for adding the peripheral blood drawn from the patient;

[0020] - A CTC circulating tumor cell separation device, used to separate CTC circulating tumor cells from the added peripheral blood; preferably, the CTC circulating tumor cell separation device is located in the housing;

[0021] - A CTC circulating tumor cell detection device, used to detect the separated CTC circulating tumor cells; preferably, the CTC circulating tumor cell detection device is located in the housing;

[0022] - A closable CTC circulating tumor cell culture device, used for culturing and proliferating the separated CTC circulating tumor cells in vitro; wherein the CTC circulating tumor cell culture device may be located inside the housing or outside the housing;

[0023] - A drug sensitivity detection device, used for performing drug sensitivity test on cultured CTC circulating tumor cells; preferably, the drug sensitivity detection device can be located inside the shell or outside the shell.

[0024] In a preferred embodiment of the present invention, the inner cavity of the housing is provided with a first container for receiving the peripheral blood test sample. The first container may be one or more.

[0025] More preferably, a first container holder is provided in the housing for placing the first container. More preferably, the first container holder can rotate around its rotation axis; the first container holder is provided with a plurality of first container placement slots, and one or more first container placement slots are arranged around the rotation axis.

[0026] More preferably, a first container taking-out member is provided in the housing, for taking out the first container after receiving the peripheral blood test sample from the first container placement slot, preferably, the first container can be taken out by grabbing, pushing or the like.

[0027] In a preferred embodiment of the present invention, the apparatus comprises a first container conveying device for conveying the first container to the CTC circulating tumor cell separation device. More preferably, the first container taking out member places the taken out first container to the first container conveying device.

[0028] In a preferred embodiment of the present invention, the device comprises a second container for receiving the separated CTC circulating tumor cells.

[0029] In a preferred embodiment of the present invention, the apparatus comprises a second container conveying device for conveying the second container to the CTC circulating tumor cell detection device and the CTC circulating tumor cell culture device sequentially or separately.

[0030] More preferably, the second container conveying device includes a first end removal piece for taking out the second container and placing it on the second container conveying device; and / or the second container conveying device includes a second end removal piece for taking out the second container on the second container conveying device and placing it on the CTC circulating tumor cell detection device and / or the CTC circulating tumor cell culture device.

[0031] More preferably, the CTC circulating tumor cell detection device and / or CTC circulating tumor cell culture is provided with a second container holder for placing a second container. More preferably, there are multiple second containers, and the second container holder can rotate around its rotation axis; the second container holder is provided with multiple second container placement slots, and the multiple second container placement slots are arranged around the rotation axis.

[0032] In a more preferred embodiment of the present invention, the CTC circulating tumor cell culture device and the drug sensitivity detection device are integrated into one. For example, the CTC circulating tumor cell culture device and the drug sensitivity detection device share a sealable chamber, and the sealable chamber is provided with a temperature regulator, a nitrogen dioxide concentration detector, a gas inlet and outlet, and a liquid material addition port.

[0033] In a more preferred embodiment of the present invention, the closable chamber is provided with an observation window, which may be a transparent window fixed on the closable chamber or a lens that can extend into the closable chamber. Alternatively, in another more preferred embodiment, the closable chamber is provided with a video and / or photo device. The video and / or photo device may replace the observation window or exist together with the observation window.

[0034] In a preferred embodiment of the present invention, the CTC circulating tumor cell separation device and the CTC circulating tumor cell detection device are located inside the shell, and the CTC circulating tumor cell culture device and the drug sensitivity detection device are located outside the shell and share a closable chamber.

[0035] In a more preferred embodiment of the present invention, a plurality of closable chambers are provided, and one or more CTC circulating tumor cell culture devices and the drug sensitivity detection device are arranged in each closable chamber.

[0036] In a more preferred embodiment, a transport channel is provided to connect the plurality of closable chambers for transporting the second container into a desired closable chamber.

[0037] In a preferred embodiment of the present invention, the apparatus comprises a processor and a data output device, wherein the processor operates the data output device to output the CTC circulating tumor cell detection results and / or the drug sensitivity test results, or the processor is capable of converting the CTC circulating tumor cell detection results into a disease condition and / or is capable of converting the drug sensitivity test results into a clinical medication guidance plan for the patient.

[0038] In a preferred embodiment of the present invention, the device also includes a storage device for storing the test data from each peripheral blood sample of the patient. Preferably, the test data includes at least any one or more of the following data: CTC circulating tumor cell detection results, disease status, CTC circulating tumor cell resistance, medication regimen, and number of repeated tests.

[0039] In a preferred embodiment of the present invention, before and / or after each peripheral blood sample test, the processor reads the test data of the previous peripheral blood sample of the same patient, and compares and / or outputs the data together with the previous peripheral blood sample.

[0040] In a preferred embodiment of the present invention, the volume of the first container is not less than 5 ml, preferably not less than 5 ml, preferably 5-10 ml, more preferably 6-9 ml, preferably 7-8.5 ml, generally 7.5-8 ml.

[0041] The second aspect of the present invention is to provide a method for accurately treating tumors based on CTC circulating tumor cells, comprising the following steps:

[0042] Step 1: Draw peripheral blood from the patient;

[0043] Step 2: Isolation of CTC circulating tumor cells;

[0044] Step 3: Evaluate the disease status through CTC circulating tumor cell detection results;

[0045] Step 4: CTC circulating tumor cells culture and proliferation in vitro;

[0046] Step 5: Parallel drug sensitivity test, and select the best medication regimen based on the drug resistance of CTC circulating tumor cells;

[0047] Step 6: Use the selected medication regimen to guide the patient's clinical medication;

[0048] Step 7: After one course of treatment, draw peripheral blood from the patient;

[0049] Step 8: Isolate CTC circulating tumor cells;

[0050] Step 9: Evaluation. If CTC circulating tumor cells are detected, repeat steps 4 to 9. If CTC circulating tumor cells are not detected, continue to use the clinical medication regimen until CTC circulating tumor cells are no longer detected after a period of time.

[0051] Wherein, in said step 1, the amount of peripheral blood drawn from the patient is not recommended to be less than 5 ml, preferably not less than 5 ml, preferably 5-10 ml, more preferably 6-9 ml, preferably 7-8.5 ml, generally 7.5-8 ml.

[0052] In a preferred embodiment of the second aspect of the present invention, the method is implemented using the device described in the first aspect.

[0053] In a preferred embodiment of the second aspect of the present invention, the method comprises:

[0054] Step 1: Draw peripheral blood from a patient; and add the peripheral blood sample to one or more first containers;

[0055] Step 2: Isolating CTC circulating tumor cells from the peripheral blood sample in the first container, and adding the separated CTC circulating tumor cells to the second container;

[0056] Step 3: Evaluate the disease status through CTC circulating tumor cell detection results;

[0057] Step 4: sending the second container into the CTC circulating tumor cell culture device to culture and proliferate the CTC circulating tumor cells in vitro;

[0058] Step 5: Parallel drug sensitivity test, and select the best medication regimen based on the drug resistance of CTC circulating tumor cells;

[0059] Step 6: Use the selected medication regimen to guide the patient's clinical medication;

[0060] Step 7: After one course of treatment, draw peripheral blood from the patient;

[0061] Step 8: Isolate CTC circulating tumor cells;

[0062] Step 9: Evaluation. If CTC circulating tumor cells are detected, repeat steps 4 to 9. If CTC circulating tumor cells are not detected, continue to use the clinical medication regimen until CTC circulating tumor cells are no longer detected after a period of time.

[0063] In a preferred embodiment of the present invention, the method for separating CTC circulating tumor cells can be: separation by physical properties, separation by biological properties, or a combination of these two separation methods. For example, CTC circulating tumor cells can be separated from other cells in the blood by physical properties such as volume, density, charge and structure.

[0064] Among them, the separation method based on physical properties includes: 1) Leukocytes are smaller than CTC circulating tumor cells, so a filter with a pore size of 8μm can be used to remove leukocytes. 2) Ficoll (polysucrose) or other similar density gradient media can be used to separate CTC circulating tumor cells from red blood cells and monocytes in plasma. 3) Because CTC circulating tumor cells have a higher volume and deformation resistance than other cells in the blood, CTC circulating tumor cells can be captured using a microchip and then separated by dielectric electrophoresis (DEP) based on the charge properties of CTC circulating tumor cells.

[0065] Among them, the separation methods based on biological characteristics include positive selection and negative selection, which can be achieved by immunological methods or microfluidic methods. Antibody positive selection uses epithelial markers such as EpCAM, anti-mesenchymal markers such as N-cadherin, or anti-epithelial and mesenchymal markers such as plastin3. Negative selection uses anti-CD45 antibodies to remove white blood cells.

[0066] In addition, it is preferred to use a mixture of antibodies optimized for different tumor types during separation to ensure that as many tumor cells as possible are obtained. For example, QIAGEN's Adna Test CancerSelect uses magnetic beads combined with antibodies to separate tumor cells from blood samples of tumor patients.

[0067] In a preferred embodiment of the present invention, the CTC circulating tumor cell detection result includes at least the number (number) of CTC circulating tumor cells, characteristic types, etc. The CTC circulating tumor cell detection result may also include activity detection of the separated CTC circulating tumor cells to determine their activity.

[0068] In a preferred embodiment of the present invention, the culture and proliferation of CTC circulating tumor cells are mainly based on the conditions of simulating the environment in the patient's body as much as possible. In terms of CTC circulating tumor cell in vitro culture and proliferation technology, the main conditions are pollution-free environment, constant temperature, gas environment, cell culture solution pH concentration and cell culture medium.

[0069] The temperature of the CTC circulating tumor cell culture is preferably 36.5°C±0.5°C.

[0070] The gas environment for culturing the CTC circulating tumor cells preferably contains CO2.

[0071] In a preferred embodiment of the present invention, NaHCO3 is added to the cell culture medium of the CTC circulating tumor cells to provide CO2; hydroxyethylpiperazine ethanesulfonic acid (HEPES) can also be added to the cell culture medium, which is non-toxic to cells and also has a buffering effect and has the characteristic of preventing rapid changes in pH. When used in open cell culture technology, its greatest advantage is that it can maintain a relatively constant pH value during open culture or cell observation.

[0072] The cell culture medium for the CTC circulating tumor cell culture is a basic substance for supplying cell nutrition and promoting cell proliferation in the cultured cells, and is also a living environment for the growth and proliferation of the cultured cells. In a preferred embodiment of the present invention, the cell culture medium adopts a synthetic culture medium and / or a natural culture medium. The synthetic culture medium is strictly configured according to the type and quantity of substances required by the cells, and contains carbohydrates, amino acids, lipids, inorganic salts, vitamins, trace elements and cell growth factors. The natural culture medium adopts calf serum. Calf serum contains a variety of cell growth factors, adhesion promoting factors and their multi-active substances, which can make cells proliferate and grow smoothly when used in combination with synthetic culture medium.

[0073] In a preferred embodiment of the present invention, the drug sensitivity test is preferably a parallel drug sensitivity test, that is, a parallel sensitivity test of a group of drugs is performed on CTC circulating tumor cells that have been cultured and proliferated in vitro. The drug is recommended to use FDA-approved drugs during treatment. Drugs that have not been approved by the FDA can also be used, but are preferably used for research. The selection of the same group of drugs is open, including various chemotherapeutic drugs, targeted drugs, and biological agents, etc., and the same group can be different drugs of the same type, or different types of drug combinations. The drug sensitivity test can be carried out using adenosine triphosphate bioluminescence method (ATP-TCA) and collagen gel droplet embedding culture drug sensitivity test method (CD-DST).

[0074] The preferred medication regimen can be obtained according to different test methods mentioned above. Generally, the medication regimen to which the cells are sensitive to the drug is preferred, which can be a single drug or a combination of several drugs. The medication regimen with small toxic and side effects is more preferred.

[0075] The drugs used in the parallel drug sensitivity test are preferably selected from the group consisting of: nitrogen mustard, nitrogen methyl, cyclophosphamide, ifosfamide, chlorambucil, thiotepa, busulfan, hexamethylmelamine, melphalan, carmustine, semustine, fotemustine, nimustine, lomustine, nitrocarmustine, cisplatin, carboplatin, oxaliplatin, methotrexate, fluorouracil, tegafur, uracil tegafur, tegafur-uracil, doxifluridine, carmoflurane, capecitabine, mercaptopurine, thioguanine, hydroxyurea, cytarabine, gemcitabine, fludarabine, aclarubicin, amazine, pirarubicin, epirubicin, doxorubicin, actinomycin D, mitoxantrone, daunorubicin, mitomycin, bleomycin, bleomycin, bleomycin, idarubicin, irinotecan, hydroxycamptothecin, vindesine, vinblastine, choline, vincristine, paclitaxel, homoharringtonine, cantharidin, norcantharidin, asparaginase, pegaspargase, anastrozole, aminoglutethimide, nilutamide, nandrolone phenylpropionate, bicalutamide, testosterone propionate, buserelin, danazol, dexamethasone, flutamide, formestane, testosterone esters, leuprolide, gonadorelin, goserelin, nafarelin, triptorelin , progesterone, diethylstilbestrol, megestrol acetate, methyltestosterone, methylprednisolone, medroxyprogesterone, thyroid tablets, levothyroxine sodium, letrozole, mitotane, chlorethoxyquin, spironolactone, prednisone, prednisolone, hydrocortisone, ethinyl estradiol, tamoxifen, exemestane, basiliximab, bevacizumab, rituximab, panitumumab, trastuzumab, ibritumomab tiuxetan, cetuximab Monoclonal antibody, bortezomib, lapatinib, sunitinib, interleukin-2, recombinant interferon α, recombinant interferon β-1α, recombinant human adenovirus, bexarotene, dasatinib, indirubin, erlotinib, fulvestrant, gefitinib, mesodrin, lenalidomide, clofazimine, clodronate disodium, chloroquine, miltefosine, pamidronate disodium, pemetrexed, arsenic trioxide, lycorine, sorafenib, retinoic acid, thymosin, thymopentin, endostatin, ibandronic acid, imatinib, zoledronic acid, abarelix, abiraterone acetate, trastuzumab-Ematansine conjugate, afatinib, alemtuzumab, aretinoin, Erwinia chrysanthemi asparaginase, axitinib, azacitidine, bendamustine hydrochloride, besaro Ding, bosutinib, cabazitaxel, cabozantinib, carfilzomib, choline C11, cinacalcet hydrochloride, clofarabine, dabrafenib, degarelix, denileukin 2, denosumab, enzalutamide, eribulin mesylate, glutaraldehyde, ibrutinib, ipilimumab, ixabepilone, mitotane, nelarabine, 99-technetium-labeled anticancer monoclonal antibody, ofam Monoclonal antibody, mebutazine, panitumumab, pazopanib, pegastatin, pentostatin, pertuzumab, pomalidomide, ponatinib, porfimer sodium, pralatrexate, radium-223 chloride, rasburicase, regorafenib, romidepsin, adenosine, streptozocin, temsirolimus, trametinib, valrubicin, vandetanib, vemurafenib, vismodegib, vorinostat, aflibercept,Tbo-filgrastim, Technetium 9Tc 99m tilmanocept, Obinutuzumab, Brentuximab Bedotin, Buffered Intrathecal Electrolyte / Dextrose. ,

[0076] It should be understood that the selection of the above drugs is preferably based on drug sensitivity tests with reference to the already defined indications.

[0077] The device provided by the present invention has the following advantages:

[0078] 1. Using CTC circulating tumor cells for tumor treatment and evaluation has little impact on patients (only 5-10 ml of peripheral blood is drawn from the patient).

[0079] 2. The in vitro drug sensitivity test is conducted using the patient's CTC circulating tumor cells, which has a personalized and precise treatment effect for the patient.

[0080] 3. The method of parallel drug sensitivity testing using different clinical drugs can optimize a clinical medication regimen each time, so that tumor cells are always facing attacks from different types of drugs, thereby weakening the tumor's ability to adapt to drugs and avoiding the drug resistance problem caused by a single clinical drug.

[0081] 4. It can realize cyclic treatment steps and open clinical drug selection methods, making this treatment method sustainable and developable.

[0082] 5. Combining treatment with evaluation, and with a shorter evaluation time, it is beneficial to quickly screen clinical medications for patients and enter personalized treatment;

[0083] 6. The device described in the present invention can realize the process of disease assessment and screening of clinical drugs, and preferably can realize the detection and report output of multiple patient samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 This is a schematic diagram of the external structure of a device for a precise tumor treatment method based on CTC circulating tumor cells in one embodiment of the present invention;

[0085] Figure 2 This is a schematic diagram of the structure of a first container support or a second container support in an embodiment of the present invention;

[0086] Figure 3 This is a schematic diagram of transporting the first container and the second container of a device for a precise tumor treatment method based on CTC circulating tumor cells in one embodiment of the present invention. Specific embodiments

[0088] like Figure 1 and 3 As shown, the device for the precise treatment method of tumors based on CTC circulating tumor cells described in the present invention includes a shell 1, a peripheral blood sample injection hole 12 and a data display screen 11 are provided on the wall of the shell 1, and a processor (not shown), a CTC circulating tumor cell separation device 6, and a first container bracket 2 are provided in the shell.

[0089] like Figure 2 and Figure 3 As shown, the first container holder 2 includes a disc-shaped or cylindrical main body 21 , the center of the main body 21 is a rotation axis 23 , and first container placement grooves 22 are distributed around the rotation axis 23 , and a plurality of first containers 20 are placed in the first container placement grooves 22 .

[0090] The peripheral blood sample extracted from the patient is added into the first container 20 through the housing 1. Sometimes, when multiple samples are needed, after each first container 20 is added, the first container holder 2 rotates around the rotation axis 23, and the next first container 20 is placed under the peripheral blood sample addition hole 12, and subsequent samples are gradually added.

[0091] After the sample addition is completed, the gripper 72 places the first containers 20 containing the peripheral blood samples one by one on the first container conveying device 3. Figure 3 A gripping device including two grippers 72 is shown, and the two grippers 72 are connected by a rotating arm 71, but it can also be one or more grippers. When the rotating arm 71 rotates, the multiple grippers 72 will work alternately to take out the first container 20 and place it into the first container conveying device 3. The first container conveying device 3 conveys the first container into the CTC circulating tumor cell separation device 6 to separate the CTC circulating tumor cells from the peripheral blood sample.

[0092] There are many methods for isolating CTCs, but they can generally be divided into two categories: separation by physical properties and separation by biological properties. In practice, most separation methods combine these two principles.

[0093] Separation by physical properties can be done by separating CTCs from other cells in the blood by physical properties such as size, density, charge, and structure. Leukocytes are smaller than CTCs, so 8μm pore filters can be used to remove them. Ficoll (polysucrose) or other similar density gradient media can be used to separate CTCs from red blood cells and monocytes in plasma. Because CTCs have a higher volume and resistance to deformation than other cells in the blood, they can be captured using a microchip and then separated by dielectric electrophoresis (DEP) based on the charge properties of the CTCs.

[0094] The biological characteristics of CTC separation include positive selection and negative selection, which can be achieved by immunological methods or microfluidic methods. Antibody positive selection uses epithelial markers such as EpCAM, anti-mesenchymal markers such as N-cadherin, or anti-epithelial and mesenchymal markers such as plastin3. Negative selection uses anti-CD45 antibodies to remove white blood cells. Regardless of the method used, the operating protocol must be strictly followed when isolating and capturing such small numbers of cells.

[0095] In addition, a mixture of antibodies optimized for different tumor types should be used during separation to ensure that as many tumor cells as possible are obtained. For example, QIAGEN's Adna Test CancerSelect uses magnetic beads combined with antibodies to separate tumor cells from blood samples of tumor patients.

[0096] The separated CTC circulating tumor cells are placed in a plurality of second containers 511, wherein the samples of one or more second containers 511 are passed through a CTC circulating tumor cell detection device (not shown). The CTC circulating tumor cell detection items include the number (number) of CTCs, characteristic types, etc., which can be performed directly in the CTC circulating tumor cell separation device 6, or can be transported to a separate CTC circulating tumor cell detection device for detection. The processor outputs the detection and evaluation results through a display screen 11, or may also include a printer, etc., to output the results.

[0097] With the advancement of technology, CTC circulating tumor cell detection technology can be used to study the molecular characteristics of CTC. By comparing the molecular characteristics of CTC before and after treatment, the patient's disease course can be analyzed and studied, such as the drug resistance of CTC circulating tumor cells. The evaluation also includes activity research on the isolated CTC circulating tumor cells to determine their activity.

[0098] The remaining second containers 511 are delivered to the CTC circulating tumor cell culture device 5 through the second container delivery device 4. The CTC circulating tumor cell culture device 5 may be composed of a plurality of units, and each patient's sample uses a separate unit, such as Figure 1 As shown, at least three units 51 , 52 and 53 are included, and the units are connected by a transmission device 50 . The second container 511 is delivered to the desired unit through the transmission device 50 .

[0099] like Figure 3 As shown, a second container holder 510 is provided in the CTC circulating tumor cell culture device 5, and its structure is the same or similar to that of the first container holder, including a rotation axis 513 at the center and second container placement slots 512 distributed around the rotation axis 513. The gripper 72 takes out the second container 511 and places it in the second container placement slot 512.

[0100] The main conditions in the CTC circulating tumor cell culture device 5 are to simulate the environment in the patient's body as much as possible. In terms of CTC cell in vitro culture and proliferation technology, the main conditions are pollution-free environment, constant temperature, gas environment, cell culture solution pH concentration and cell culture medium.

[0101] A pollution-free environment, a non-toxic and sterile culture environment are the primary conditions for ensuring the survival of CTC cells. When CTC cells are placed in vitro for culture, they lose their ability to defend against microorganisms and toxic substances compared to in vivo. Once they are contaminated or their own metabolites accumulate, they may die. Therefore, during the culture, keeping the cell living environment pollution-free and removing metabolites in a timely manner are basic conditions for maintaining cell survival.

[0102] Constant temperature: To maintain vigorous growth of cultured cells, a constant and suitable temperature is necessary. The optimum temperature for culturing cells is equivalent to the normal temperature of the body from which various cells or tissues are taken. The standard temperature for CTC cell culture is 36.5℃±0.5℃. If it deviates from this temperature range, the normal metabolism of cells will be affected, or even die.

[0103] Gas environment: Gas is a necessary condition for the survival of CTC cell culture. The gases required are mainly oxygen and carbon dioxide. Oxygen participates in the tricarboxylic acid cycle, produces energy for cell growth and proliferation, and synthesizes various components required for cell growth. During open culture, CTC cells are placed in a mixed gas environment of 95% air and 5% carbon dioxide. Carbon dioxide is both a cell metabolic product and a component required for cell growth and reproduction. Its main role in cell culture is to maintain the pH value of the culture medium.

[0104] PH concentration of cell culture medium: adding NaHCO3 can provide CO2, but CO2 is easy to escape, so it is most suitable for closed culture; while hydroxyethylpiperazine ethanesulfonic acid (HEPES) is non-toxic to cells and also has a buffering effect, which has the characteristic of preventing rapid changes in pH. It is used in open cell culture technology. Its biggest advantage is that it can maintain a relatively constant pH value during open culture or cell observation.

[0105] Cell culture medium: The culture medium is the basic substance that provides cell nutrition and promotes cell proliferation in cultured cells. It is also the living environment for the growth and proliferation of cultured cells. The culture medium uses synthetic culture medium and natural culture medium. Synthetic culture medium is strictly configured according to the types and quantities of substances required by cells, and contains carbohydrates, amino acids, lipids, inorganic salts, vitamins, trace elements and cell growth factors. Natural culture medium uses calf serum. Calf serum contains a variety of cell growth factors, adhesion promoting factors and their multi-active substances. When used in combination with synthetic culture medium, it can make cells proliferate and grow smoothly.

[0106] During and after the culture, the cell culture and proliferation conditions are observed by video and / or photo equipment or through an observation window.

[0107] After the culture is completed, referring to the peripheral blood sample loading method, the CTC circulating tumor cell culture device 5 is provided with a drug loading hole, through which drugs are added to the second container 511. The second container holder 510 rotates, and each second container 511 is placed under the drug loading hole one by one, and different drugs are added to each second container 511, and parallel drug sensitivity tests are performed to observe the results of the drug sensitivity tests.

[0108] The device of the present invention may also include a storage device to store information such as drug sensitivity test results, disease conditions, and previously used drugs (especially drugs to which drug resistance has developed). Before each drug sensitivity test, the previous information is read to avoid repeated drug sensitivity tests. After the experiment is completed, the information can also be compared with the previous information to determine the treatment situation and select a better medication plan.

[0109] The parallel drug sensitivity test is a parallel sensitivity test of a group of drugs on CTC cells that have been proliferated by in vitro culture. The drugs approved by the FDA are recommended for use during treatment, and drugs not approved by the FDA are only for research. The selection of the same group of drugs is open, including various chemotherapeutic drugs, targeted drugs and biological agents, etc., and the same group can be different drugs of the same type, or different types of drug combinations. The drug sensitivity test can be carried out using adenosine triphosphate bioluminescence method (ATP-TCA) and collagen gel droplet embedding culture drug sensitivity test method (CD-DST). The preferred parallel test scheme can be obtained according to different above-mentioned test methods.

[0110] Example 1

[0111] Using the device of the present invention, the present invention also provides a method for accurately treating tumors based on CTC circulating tumor cells, comprising the following steps:

[0112] Step 1: Draw 5-10 ml of peripheral blood from a breast cancer patient

[0113] The patient's peripheral blood is drawn 5-10 ml, generally 7.5 ml, and less than 5 ml is not recommended. The storage and transportation of blood must be carried out within the recommended temperature range, i.e., refrigerated at 4°C to 10°C, or at room temperature of 15°C to 30°C. The blood cannot be frozen and is added to the first container.

[0114] Step 2: Isolation of circulating tumor cells (CTCs)

[0115] The separation of CTC circulating tumor cells should be completed within a specified time after blood drawing, which generally varies according to the CTC isolation method and may be several hours or several days. If the specified time is exceeded, some blood samples may coagulate and agglomerate even in the presence of anticoagulants.

[0116] The separated CTC circulating tumor cells are added to the second container.

[0117] Step 3: Evaluation

[0118] Whether before or after treatment, the number of CTCs is the main reference indicator. Clinical studies have confirmed that when the CTCs in the blood sample are higher than the cut-off (≥5 / 7.5ml), it indicates a poor prognosis, and when the CTCs in the sample are lower than the cut-off, it indicates a good prognosis. More importantly, after treatment, comparing the number of CTCs before treatment will help determine the progression of the patient's disease or the treatment status.

[0119] An increase in the number of CTCs indicates a poor therapeutic effect, while a decrease in the number of CTCs indicates a good therapeutic effect. This real-time, sensitive, and reliable evaluation method can help doctors correctly judge the efficacy and develop efficient individualized treatment plans. CTC evaluation can judge the prognosis earlier, more accurately, and more sensitively than traditional imaging. A large number of studies have shown that the results of CTC monitoring for prognosis judgment after 4 weeks of treatment are extremely consistent with the results of traditional imaging after 12 weeks of treatment. Moreover, when the CTC value is greater than or equal to the cut-off value, CTC evaluation is more accurate than imaging for prognosis judgment.

[0120] The evaluation results are output through the display screen for the doctor's reference.

[0121] Step 4: CTC circulating tumor cells culture and proliferation in vitro

[0122] The CTC circulating tumor cells in the second container are sent to the CTC circulating tumor cell culture device for in vitro culture and proliferation.

[0123] Step 5: Parallel drug sensitivity test, choose one of the options

[0124] The information in the storage device shows that the patient has developed resistance to cyclophosphamide, ifosfamide, and exemestane. Therefore, the above three drugs were excluded during the drug sensitivity test, and the following drugs were selected for parallel drug sensitivity testing: thiotepa, melphalan, methotrexate, fluorouracil, uracil tegafur, doxifluridine, tegafur, carmofur, tegafur-uracil, capecitabine, aclarubicin, pirarubicin, epirubicin, doxorubicin, mitoxantrone, mitomycin, bleomycin, Vindesine, vinblastine, vincristine, paclitaxel, anastrozole, aminoglutethimide, nandrolone phenylpropionate, buserelin, formestane, testosterone complex esters, gonadorelin, goserelin, progesterone, diethylstilbestrol, megestrol acetate, medroxyprogesterone acetate, letrozole, leuprolide, prednisolone, hydrocortisone, triptorelin, ethinyl estradiol, tamoxifen, toremifene, lapatinib, rituximab, trastuzumab, interleukin-2, fulvestrant, recombinant interferon beta-1alpha, chloroquine.

[0125] The results of drug sensitivity tests showed that vincristine, paclitaxel, and recombinant interferon β-1α had the best effects in inhibiting tumor cell growth.

[0126] Step 6: Use the preferred parallel drug sensitivity test scheme to guide clinical medication for patients

[0127] The clinical medication regimen for guiding patients is based on the preferred regimen obtained in step 5, i.e., vincristine, paclitaxel and recombinant interferon β-1α are selected as therapeutic drugs. The doctor selected paclitaxel as the therapeutic drug, and the medication guidance is carried out with reference to the instructions for use of the selected clinical drug.

[0128] Step 7: After one course of treatment, draw 5-10 ml of peripheral blood from the patient

[0129] The duration of one course of treatment is determined by referring to the instructions for use of the clinical drug selected in step 6. The amount of 5-10 ml of peripheral blood drawn from the patient should be consistent with the amount of peripheral blood drawn from the patient last time, such as 7.5 ml. The uniform amount of peripheral blood drawn is used to provide a uniform comparison condition for CTC evaluation before and after treatment.

[0130] Step 8: Isolation of circulating tumor cells (CTCs)

[0131] The method for isolating CTC circulating tumor cells is the same as step 2, using the same separation method and conditions so that there is a unified comparison condition for CTC evaluation before and after treatment.

[0132] Step 9: Evaluation

[0133] The evaluation method and conditions described in step 9 are all carried out in accordance with step 3, which is also to unify the evaluation method. If CTC is not detected during the evaluation for a period of time, it can be judged that the treatment is progressing well, indicating that the patient's tumor spread is under control. The period of time described depends on the condition and the drug, and is generally three consecutive courses of the clinical drug used. That is, under normal circumstances, if CTC circulating tumor cells are not detected during the evaluation after three consecutive courses of clinical drugs used, it can be judged that the condition is under stable control. Once CTC is detected, the above step 4 will be repeated to enter the CTC circulating tumor cell in vitro culture and proliferation step, and then continue all the processes of steps 5-9 to enter the diagnosis and treatment cycle. If CTC is not detected once or twice, the preferred clinical medication regimen can continue to be used.

[0134] No CTC was detected in this patient, so the previous medication regimen can be continued.

[0135] Example 2

[0136] The second embodiment of the method for precise tumor treatment based on CTC circulating tumor cells of the present invention comprises the following steps:

[0137] Step 1: Draw 5-10 ml of peripheral blood from prostate cancer patients

[0138] The patient's peripheral blood is drawn 5-10 ml, generally 7.5 ml, and less than 5 ml is not recommended. The storage and transportation of blood must be carried out within the recommended temperature range, i.e. 4°C to 10°C refrigeration, or room temperature 15°C to 30°C, and the blood cannot be frozen.

[0139] Step 2: Isolation of circulating tumor cells (CTCs)

[0140] The separation of CTC circulating tumor cells should be completed within a specified time after blood drawing, which generally varies according to the CTC isolation method and may be several hours or several days. If the specified time is exceeded, some blood samples may coagulate and agglomerate even in the presence of anticoagulants.

[0141] Step 3: Evaluation

[0142] Clinical studies have confirmed that when the CTC in the blood sample is higher than the cut-off (≥5 / 7.5ml), it indicates a poor prognosis, and when the CTC in the sample is lower than the threshold (cut-off), it indicates a good prognosis. More importantly, when treatment is taken, comparing the number of CTCs before treatment will help determine the progression of the patient's disease or treatment status.

[0143] An increase in the number of CTCs indicates a poor therapeutic effect, while a decrease in the number of CTCs indicates a good therapeutic effect. This real-time, sensitive, and reliable evaluation method can help doctors correctly judge the efficacy and develop efficient individualized treatment plans. CTC evaluation can judge prognosis earlier, more accurately, and more sensitively than traditional imaging. A large number of studies have shown that the results of CTC monitoring for prognosis after 4 weeks of treatment are extremely consistent with the results of traditional imaging after 12 weeks of treatment. Moreover, when the CTC value is greater than or equal to the cut-off value, CTC evaluation is more accurate than imaging for prognosis judgment. CTC evaluation is better at judging efficacy and prognosis than a single biological marker. Studies have shown that in prostate cancer, CTC evaluation is more accurate and sensitive for prognosis judgment than the traditional marker PSA.

[0144] Step 4: CTC circulating tumor cells culture and proliferation in vitro

[0145] Step 5: Parallel drug sensitivity test, choose one of the options

[0146] The following drugs were selected for parallel drug sensitivity testing: cisplatin, tegafur, carmofur, doxorubicin, mitoxantrone, buserelin, danazol, flutamide, gonadorelin, goserelin, progesterone, diethylstilbestrol, megestrol acetate, medroxyprogesterone acetate, flutriethoxyfen, nafarelin, nilutamide, prednisolone, hydrocortisone, triptorelin, ethinyl estradiol, and tamoxifen.

[0147] Among them, the combination of buserelin and tamoxifen has the best effect.

[0148] Step 6: Use the preferred parallel drug sensitivity test scheme to guide clinical medication for patients

[0149] The clinical medication regimen for guiding patients is based on the preferred regimen obtained in step 5, i.e., buserelin and tamoxifen are used together. The medication guidance is carried out with reference to the instructions for use of the selected clinical drugs.

[0150] Step 7: After one course of treatment, draw 5-10 ml of peripheral blood from the patient

[0151] The duration of one course of treatment is determined by referring to the instructions for use of the clinical drug selected in step 6. The amount of 5-10 ml of peripheral blood drawn from the patient should be consistent with the amount of peripheral blood drawn from the patient last time, such as 7.5 ml. The uniform amount of peripheral blood drawn is used to provide a uniform comparison condition for CTC evaluation before and after treatment.

[0152] Step 8: Isolation of circulating tumor cells (CTCs)

[0153] The method for isolating CTC circulating tumor cells is the same as step 2, using the same separation method and conditions so that there is a unified comparison condition for CTC evaluation before and after treatment.

[0154] Step 9: Evaluation

[0155] (1) If CTCs are not detected for a period of time, the condition is under control and the patient enters the observation state; (2) If CTCs are detected, repeat step 4 and subsequent steps above; (3) If CTCs are not detected, continue to use the clinical medication plan.

[0156] The evaluation method and conditions described in step 9 are all carried out in accordance with step 3, which is also to unify the evaluation method. If CTC is not detected during the evaluation for a period of time, it can be judged that the treatment is progressing well, indicating that the patient's tumor spread is under control. The period of time described depends on the condition and the drug, and is generally three consecutive courses of the clinical drug used. That is, under normal circumstances, if CTC circulating tumor cells are not detected during the evaluation after three consecutive courses of clinical drugs used, it can be judged that the condition is under stable control. Once CTC is detected, the above step 4 will be repeated to enter the CTC circulating tumor cell in vitro culture and proliferation step, and then continue all the processes of steps 5-9 to enter the diagnosis and treatment cycle. If CTC is not detected once or twice, the preferred clinical medication regimen can continue to be used.

[0157] No CTC circulating tumor cells were detected in this patient, and the previous treatment regimen was continued.

[0158] Example 3

[0159] The third embodiment of the method for precise tumor treatment based on CTC circulating tumor cells of the present invention comprises the following steps:

[0160] Step 1: Draw 5-10 ml of peripheral blood from colon cancer patients

[0161] The patient's peripheral blood is drawn 5-10 ml, generally 7.5 ml, and less than 5 ml is not recommended. The storage and transportation of blood must be carried out within the recommended temperature range, i.e. 4°C to 10°C refrigeration, or room temperature 15°C to 30°C, and the blood cannot be frozen.

[0162] Step 2: Isolation of circulating tumor cells (CTCs)

[0163] The separation of CTC circulating tumor cells should be completed within a specified time after blood drawing, which generally varies according to the CTC isolation method and may be several hours or several days. If the specified time is exceeded, some blood samples may coagulate and agglomerate even in the presence of anticoagulants.

[0164] Step 3: Evaluation

[0165] Whether before or after treatment, the number of CTCs is the main reference indicator. Clinical studies have confirmed that when the CTCs in the blood sample are higher than the cut-off (≥3 / 7.5ml), it indicates a poor prognosis, and when the CTCs in the sample are lower than the threshold (cut-off), it indicates a good prognosis. More importantly, after treatment, comparing the number of CTCs before treatment will help determine the progression of the patient's disease or treatment status.

[0166] An increase in the number of CTCs indicates a poor therapeutic effect, while a decrease in the number of CTCs indicates a good therapeutic effect. This real-time, sensitive, and reliable evaluation method can help doctors correctly judge the efficacy and develop efficient individualized treatment plans. CTC evaluation can judge the prognosis earlier, more accurately, and more sensitively than traditional imaging. A large number of studies have shown that the results of CTC monitoring for prognosis judgment after 4 weeks of treatment are extremely consistent with the results of traditional imaging after 12 weeks of treatment. Moreover, when the CTC value is greater than or equal to the cut-off value, CTC evaluation is more accurate than imaging for prognosis judgment.

[0167] Step 4: CTC circulating tumor cells culture and proliferation in vitro

[0168] Step 5: Parallel drug sensitivity test, choose one of the options

[0169] The patient was resistant to oxaliplatin, so this drug was excluded. The following drugs were selected for parallel drug sensitivity testing: nimustine, doxifluridine, tegafur, carmofur, tegafur-uracil, capecitabine, epirubicin, mitomycin, bevacizumab, panitumumab, and cetuximab.

[0170] Among them, tegafur-uracil, panitumumab, mitomycin, and capecitabine have better effects in inhibiting tumor cell growth.

[0171] Step 6: Use the preferred parallel drug sensitivity test scheme to guide clinical medication for patients

[0172] The clinical medication regimen for guiding patients is based on the preferred regimen obtained in step 5, i.e., selecting tegafur-uracil, panitumumab, mitomycin, and capecitabine. In the end, the doctor selected mitomycin for treatment, and the medication guidance was carried out with reference to the instructions for use of the selected clinical drug.

[0173] Step 7: After one course of treatment, draw 5-10 ml of peripheral blood from the patient

[0174] The duration of one course of treatment is determined by referring to the instructions for use of the clinical drug selected in step 6. The amount of 5-10 ml of peripheral blood drawn from the patient should be consistent with the amount of peripheral blood drawn from the patient last time, such as 7.5 ml. The uniform amount of peripheral blood drawn is used to provide a uniform comparison condition for CTC evaluation before and after treatment.

[0175] Step 8: Isolation of circulating tumor cells (CTCs)

[0176] The method for isolating CTC circulating tumor cells is the same as step 2, using the same separation method and conditions so that there is a unified comparison condition for CTC evaluation before and after treatment.

[0177] Step 9: Evaluation

[0178] (1) If CTCs are not detected for a period of time, the condition is under control and the patient enters the observation state; (2) If CTCs are detected, repeat step 4 and subsequent steps above; (3) If CTCs are not detected, continue to use the clinical medication plan.

[0179] The evaluation method and conditions described in step 9 are all carried out in accordance with step 3, which is also to unify the evaluation method. If CTC is not detected during the evaluation for a period of time, it can be judged that the treatment is progressing well, indicating that the patient's tumor spread is under control. The period of time described depends on the condition and the drug, and is generally three consecutive courses of the clinical drug used. That is, under normal circumstances, if CTC circulating tumor cells are not detected during the evaluation after three consecutive courses of clinical drugs used, it can be judged that the condition is under stable control. Once CTC is detected, the above step 4 will be repeated to enter the CTC circulating tumor cell in vitro culture and proliferation step, and then continue all the processes of steps 5-9 to enter the diagnosis and treatment cycle. If CTC is not detected once or twice, the preferred clinical medication regimen can continue to be used.

[0180] After three courses of treatment, the patient's CTC circulating tumor cells still existed, so the parallel drug sensitivity test was continued. The patient had a certain degree of resistance to mitomycin and gave up the drug. Through the parallel drug sensitivity test, the doctor chose capecitabine as the treatment drug. After three courses of treatment, the CTC circulating tumor cells decreased significantly.

[0181] Since drug sensitivity tests have been conducted before the use of the drugs of the present invention, ineffective medication caused by drug resistance can be avoided to the greatest extent. Experiments show that except for a very small number of tumor patients with severe drug resistance, >99.9% of patients can find accurate treatment methods through the method of the present invention. Among them, patients in the middle and late stages with partial drug resistance have obtained good treatment effects through the method of the present invention. Compared with traditional chemotherapy methods, the survival period is extended by 25-36%. At the same time, the toxic and side effects of the drug are significantly reduced, which significantly improves the quality of life.

[0182] The specific embodiments of the present invention are described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions made to the present invention are also within the scope of the present invention. Therefore, the equalization changes and modifications made without departing from the spirit and scope of the present invention should be included in the scope of the present invention.

Claims

1. A device for a precise tumor treatment method based on CTC circulating tumor cells, characterized in that: include: - a housing, wherein the housing is provided with a hole for adding the peripheral blood drawn from the patient; ——CTC circulating tumor cell separation device, used to separate CTC circulating tumor cells from the added peripheral blood; ——CTC circulating tumor cell detection equipment, used to detect isolated CTC circulating tumor cells; ——A closable CTC circulating tumor cell culture device, used to culture and proliferate the isolated CTC circulating tumor cells in vitro; ——Drug sensitivity testing equipment, used to conduct drug sensitivity tests on cultured CTC circulating tumor cells; - A processor and a data output device, wherein the processor controls the data output device to output the CTC circulating tumor cell detection results and / or the drug sensitivity test results, or the processor can convert the CTC circulating tumor cell detection results into a disease condition and / or can convert the drug sensitivity test results into a patient's clinical medication guidance plan for output; - A storage device for storing the test data of each peripheral blood sample from the patient. Before and / or after each peripheral blood sample test, the processor reads the test data of the previous peripheral blood sample of the same patient, and compares and / or outputs the test data together with the previous peripheral blood sample; - A second container for receiving the separated CTC circulating tumor cells; - A second container conveying device, used for conveying the second container to the CTC circulating tumor cell detection device and the CTC circulating tumor cell culture device in sequence or separately; the inner cavity of the shell is provided with a first container for receiving the peripheral blood test sample, the shell is provided with a first container holder for placing the first container, and the first container holder can rotate around its rotation axis; the first container holder is provided with a plurality of first container placement slots, and the plurality of first container placement slots are arranged around the rotation axis; The second container conveying device includes a first end taking-out piece for taking out the second container and placing it on the second container conveying device; and / or the second container conveying device includes a second end taking-out piece for taking out the second container on the second container conveying device and placing it on the CTC circulating tumor cell detection device and / or the CTC circulating tumor cell culture device; The CTC circulating tumor cell culture device and the drug sensitivity detection device share a closable chamber, and the closable chamber is provided with a temperature regulator, a nitrogen dioxide concentration detector, a gas inlet and outlet, and a liquid material addition port; the closable chamber is provided with an observation window, or the closable chamber is provided with a video and / or photo device.

2. The device according to claim 1, characterized in that A plurality of closable chambers are provided, and one or more CTC circulating tumor cell culture devices and the drug sensitivity detection device are arranged in each closable chamber.

3. The device according to claim 2, characterized in that A transport channel is provided to connect the plurality of closable chambers and is used to transport the second container into a desired closable chamber.

Images