Saving type metastatic tumor treatment effect dose reaction model and preparation method thereof

Abstract

The invention discloses a saving-type metastatic tumor treatment effect dose response model and a preparation method thereof, and particularly provides a device for constructing the saving-type metastatic tumor treatment effect dose response model, the device comprises a metastatic tumor culture assembly, a stereotactic assembly and an image scanning device; the metastatic tumor culture assembly is used for culturing metastatic tumor tissues and normal tissue organs around the metastatic tumor tissues to obtain a metastatic tumor model; the three-dimensional positioning assembly is used for positioning a metastatic tumor model at a position in an organ; the image scanning device can carry out image scanning on the three-dimensional positioning assembly and the metastatic tumor model in the three-dimensional positioning assembly to obtain a three-dimensional image. The metastatic tumor model can be used for researching the biological behavior after the advanced malignant tumor is transferred to a certain organ and discussing the response and mechanism of the advanced cancer to treatment. The model prepared by the invention can reduce animal sacrifice caused by previous animal experiments on one hand, and can save resource loss of cell culture on the other hand.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a sparing metastatic tumor treatment effect dose-response model and a preparation method thereof. Background Art

[0002] Metastatic tumors refer to the metastasis of malignant tumors to other organs, which is a type of secondary tumor. Existing tumor cell cultures are generally divided into cell colony culture and animal in vivo culture. For malignant tumors, observation and testing in living animals will cause pain and sacrifice to the animals. Simple cell colonies can only study cell behavior, but cannot directly reflect the interactive relationship between cells and extracellular matrix, nor can they reflect the material exchange between tumor cells and the extracellular biological environment (normal tissue).

[0003] In addition, the organoid models obtained by the existing culture methods are soft and fluid, so there is no precise spatial division. In research, a group of samples cultured at one time can only be used to test one therapeutic intervention plan. Comparative tests between multiple groups of therapeutic intervention plans require repeated culture of cells or organoids, which consumes corresponding scientific research resources. Summary of the invention

[0004] The purpose of the present invention is to provide a sparing metastatic tumor treatment effect dose-response model and a preparation method thereof.

[0005] The present invention provides a device for constructing a conservation-type metastasis tumor treatment effect dose-response model, the device comprising a metastasis tumor culture component, a stereo positioning component and an image scanning device; the metastasis tumor culture component is used to culture metastasis tumor tissue and normal tissue organoids surrounding it to obtain a metastasis tumor model; the stereo positioning component is used to position the metastasis tumor model at a position within an organ; the image scanning device can perform image scanning on the stereo positioning component and the metastasis tumor model therein to obtain a three-dimensional image.

[0006] Furthermore, the metastasis tumor culture component includes an outer frame and an inner frame with openings at the upper ends, and the inner frame is provided with evenly distributed sieve holes; the area formed between the inner frame and the bottom wall is used to culture tumor organoids; and the area between the inner frame and the outer frame is used to culture normal tissue organoids.

[0007] Furthermore, the inner frame is located in the middle of the outer frame.

[0008] Furthermore, the sieve hole has a diameter of 1 mm.

[0009] Furthermore, the stereotaxic assembly includes a base, an organ frame, a first positioning bar, a second positioning bar and a metastasis model mounting groove, wherein the base is connected to the organ frame, the organ frame and the second positioning bar are both connected to the first positioning bar, and the metastasis model mounting groove is connected to the second positioning bar.

[0010] Furthermore, the second positioning bar is perpendicular to the first positioning bar 23 and can slide along the first positioning bar, and the metastasis model mounting groove is connected to the second positioning bar and can slide along the second positioning bar.

[0011] Furthermore, the stereo positioning assembly also includes a positioning block, which positions and fixes the second positioning bar and the metastasis tumor model mounting groove.

[0012] The present invention also provides a method for constructing a sparing metastatic tumor treatment effect dose-response model, using the above-mentioned device for constructing a sparing metastatic tumor treatment effect dose-response model.

[0013] The method comprises the following steps:

[0014] 1) Placing tumor organoids and normal tissue organoids in metastatic tumor culture components for culture respectively to obtain a metastatic tumor model;

[0015] 2) Conduct experiments on metastatic tumor models;

[0016] 3) The metastatic tumor model after the experiment is mounted on a stereotaxic assembly and scanned using an image scanning device to obtain a dose-response model of the sparing metastatic tumor treatment effect.

[0017] The experiment includes a drug treatment experiment or a radiosurgery experiment. The drug treatment experiment can be adding various tumor chemotherapy drug sustained-release tablets to the metastatic tumor model; the radiosurgery experiment can be an irradiation experiment using a radiosurgery device such as a gamma knife. The above experiments will form treatment-related gradient doses and biological effects.

[0018] Furthermore, the image scanning device is CT and MR.

[0019] The beneficial effects of the present invention are:

[0020] 1) The model of the present invention can be used to study the biological behavior of advanced malignant tumors after metastasis to certain organs, and to explore the response and mechanism of advanced cancer to treatment. This model can reduce the animal sacrifice caused by previous animal experiments, and save the resource loss of cultured cells.

[0021] 2) Compared with the existing cell colony culture that cannot directly reflect the interaction between cells and extracellular matrix, the organoids filled in the frame in the present application form a cell composition in a complete tissue structure, which can reflect the material exchange between cells and the extracellular biological environment. In the past, this material exchange process needed to be observed and tested in living animals, causing pain and sacrifice to the animals, but the present invention avoids this problem through the culture of organoids in the frame.

[0022] 3) The organoid model is soft and fluid, so there is no precise spatial division in the prior art. In the study, a group of samples cultured at one time can only be used for the test of one therapeutic intervention scheme, and the comparative test between multiple groups of therapeutic intervention schemes requires repeated culture of cells or organoids, which consumes corresponding scientific research resources. The model of the present invention uses the above-mentioned model construction device to perform stereotactic imaging positioning to achieve precise spatial division of the metastatic tumor model. In addition, a gradient dose of treatment (such as chemotherapy drugs or radiotherapy rays) is formed in the divided space, and the gradient dose forms different treatment schemes corresponding to different research groups, so that a cell culture and the established model meet the observation and comparison of multiple groups of research, effectively saving research resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of the structure of the metastasis tumor culture component.

[0024] Figure 2 It is a schematic diagram of the structure of the stereo positioning component. DETAILED DESCRIPTION

[0025] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0026] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0027] Example 1

[0028] The present invention provides a device for a conservation-type metastasis tumor treatment effect dose-response model, the device comprising a metastasis tumor culture component, a stereo positioning component and an image scanning device, the metastasis tumor culture component is used to culture metastasis tumor tissue and normal tissue organoids surrounding it to obtain a metastasis tumor model; the stereo positioning component is used to position the metastasis tumor model at a position within the organ; the image scanning device can perform image scanning on the stereo positioning component and the metastasis tumor model therein to obtain a three-dimensional image.

[0029] On this basis, the three-dimensional image is meshed and combined with the existing robotic sampling device or manual sampling, accurate sampling at different positions can be achieved. When the three-dimensional coordinate system is obtained by X-ray computed tomography (CT) and magnetic resonance imaging (MR) image scanning, the spatial meshing is performed, and the sampling accuracy can reach 0.1mm.

[0030] like Figure 1 As shown, the metastasis tumor culture component includes an outer frame 11 with an opening at the upper end and an inner frame 12, and the inner frame 12 is provided with evenly distributed sieve holes 13; the area formed between the inner frame 12 and the bottom wall is used to culture tumor organoids; the area between the inner frame 12 and the outer frame 11 is used to culture normal tissue organoids; the two can interact through the sieve holes, thereby simulating the tumor metastasized to the organs and obtaining a metastasis tumor model.

[0031] As a specific example, the inner frame located in the center has an outer diameter of 20mm, an inner diameter of 18mm, and a wall thickness of 1mm; the outer frame located at the periphery is 40mm in length and width, 1mm in wall thickness, 20mm in height, and 18mm in inner diameter depth. The central cube formed between the inner frame and the bottom wall is filled with cultured tumor organoids (for example, composed of lung cancer tumor cells and extracellular matrix, or other tumors such as breast cancer, intestinal cancer, etc.), and the peripheral area is filled with cultured normal tissue organoids (for example, composed of brain tissue cells and extracellular matrix, or organs such as liver, kidney, bone, etc.). This simulates a tumor that has metastasized to a certain organ (for example, brain metastasis from lung cancer, or metastatic tumors in other parts of the body).

[0032] The difference between organoids filled in the frame and cell colonies in previous studies is that cell colonies can only study cell behavior and cannot directly reflect the interaction between cells and the extracellular matrix, while organoids form a cell composition in a complete tissue structure and can reflect the material exchange between cells and the extracellular biological environment. In the past, this material exchange process needed to be observed and tested in living animals, causing pain and sacrifice to the animals.

[0033] The metastasis model involves a combination of two organoids, namely tumor tissue organoids and normal tissue organoids. The sieve holes between the two organoids (for example, the pore size is 1mm, the thickness of the partition is 1mm, and the sieve hole array on each partition surface is a 9×9 array). This partition design allows for the exchange of substances between tumor organoids and extracellular matrix organoids. The colloidal extracellular matrix in the organoid controls the passage of substances through the partition, that is, the formation of microflows through which substances pass. This simulation of the mutual influence between a tumor that has metastasized to an organ and the organ will help to carry out molecular biological experimental studies related to the growth and invasion of malignant tumors. The most important thing is that this model does not sacrifice animals at the expense of previous studies.

[0034] Therefore, this research model uses organoids to model metastatic tumors through physical microfluidics. Compared with the traditional method of using animals to establish a malignant tumor metastasis model, it eliminates harm to animals and effectively saves money in protecting animals.

[0035] like Figure 2 As shown, the stereo positioning assembly includes: a base 21, an organ frame 22, a first positioning bar 23, a second positioning bar 24, a metastasis model mounting groove 25 and a positioning block 26. The base 21 is connected to the organ frame 22 to form a frame that matches the organ shape; the first positioning bar 23 is connected to the frame, the second positioning bar 24 is perpendicular to the first positioning bar 23, and can slide along the first positioning bar 23, the metastasis model mounting groove 25 is connected to the second positioning bar 24, and can slide along the second positioning bar 24. When the second positioning bar 24 and the metastasis model mounting groove 25 are moved to the specified position respectively, the second positioning bar 24 and the metastasis model mounting groove 25 can be fixed by the positioning block 26 to avoid position deviation during the movement process. The metastasis tumor culture assembly matches the metastasis model mounting groove and can be connected to the metastasis model mounting groove.

[0036] Taking brain metastases as an example, Figure 2 As shown, the organ framework 22 can be made into a circular ring framework similar to the brain, which is convenient for simulating the location of metastatic tumors therein. If desired, the location for other positions can also be changed into the shape of similar other organs or the shape of the partial structure.

[0037] For aseptic operation, after the metastasis tumor culture component is connected to the metastasis tumor model mounting slot and fixed in the stereotaxic component, a detachable closed model frame can be added to the stereotaxic component. The closed model frame is preferably made of first-class material.

[0038] During the experiment, tumor organoids and normal tissue organoids are first placed in metastasis tumor culture components for culture to obtain metastasis tumor models. After various tumor chemotherapy drug sustained-release tablets are placed in the metastasis tumor model or it is irradiated with a radiosurgery device (such as a gamma knife), treatment-related gradient doses and biological effects will be formed.

[0039] The metastasis tumor culture component is installed on the stereotaxic component to the desired simulated position, and image scanning is performed using CT and MR to obtain a sparing metastasis tumor treatment effect dose response model.

[0040] Since organoids can be divided into sub-millimeter grid ranges in the spatial coordinate system, the spatial gradient dose formed by chemotherapy drug tablets or radiosurgery devices can be accurately spatially located through CT and MR scanning, with sub-millimeter positioning accuracy. After image positioning is completed, the treatment plan is designed, and the corresponding treatment effect dose also exhibits a sub-millimeter gradient attenuation in space. Since the gradient treatment effect dose division corresponds to the aforementioned grid division, a single modeling can obtain dozens of times more test samples than traditional modeling, greatly improving research efficiency. Compared with the traditional method of culturing cells multiple times for modeling, the metastasis tumor research model we applied for can form multiple research groups at one time, which can greatly save various types of cell culture-related scientific research resources required for modeling.

[0041] In addition, in the sampling process, the robot arm can be used to collect samples by cutting and inhaling steel wires that are suitable for Jiaodong-shaped solids. The robot can be programmed for precise sampling. According to the data information formed by the spatial positioning of the aforementioned stereotactic CT and MR images, the gradient dose partitioning of chemotherapy or radiotherapy can be programmed to obtain specimens of different groups, and then molecular biology related tests can be performed. At the same time, it can avoid the waste of research resources caused by wrong sampling due to human errors.

[0042] In summary, the entire process achieves significant savings in research resources, protects animals, and establishes an organoid model for metastasis tumor research.

[0043] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. A device for constructing a sparing metastatic tumor treatment effect dose-response model, characterized in that: The device includes a metastasis tumor culture component, a stereotaxic component and an image scanning device; the metastasis tumor culture component is used to culture metastasis tumor tissue and normal tissue organoids surrounding it to obtain a metastasis tumor model; the stereotaxic component is used to position the metastasis tumor model at a position within the organ; the image scanning device can perform image scanning on the stereotaxic component and the metastasis tumor model inside it to obtain a three-dimensional image.

2. The device according to claim 1, characterized in that The metastatic tumor culture component includes an outer frame and an inner frame with an opening at the upper end, and the inner frame is provided with evenly distributed sieve holes; the area formed between the inner frame and the bottom wall is used to culture tumor organoids; the area between the inner frame and the outer frame is used to culture normal tissue organoids.

3. The device according to claim 2, characterized in that The inner frame is located in the middle of the outer frame.

4. The device according to claim 2, characterized in that The sieve hole has a diameter of 1 mm.

5. The device according to claim 1, characterized in that The stereotactic assembly comprises a base, an organ frame, a first positioning bar, a second positioning bar and a metastasis model mounting groove, wherein the base is connected to the organ frame, the organ frame and the second positioning bar are both connected to the first positioning bar, and the metastasis model mounting groove is connected to the second positioning bar.

6. The device according to claim 5, characterized in that The second positioning bar is perpendicular to the first positioning bar 23 and can slide along the first positioning bar. The metastasis model mounting groove is connected to the second positioning bar and can slide along the second positioning bar.

7. The device according to claim 5, characterized in that The stereo positioning assembly also includes a positioning block, which is used to position and fix the second positioning bar and the metastasis tumor model mounting groove.

8. A method for constructing a sparing metastatic tumor treatment effect dose-response model, characterized in that: Use the device described in any one of claims 1-7.

9. The method according to claim 8, characterized in that The steps include: 1) Placing tumor organoids and normal tissue organoids in metastatic tumor culture components for culture respectively to obtain a metastatic tumor model; 2) Conduct experiments on metastatic tumor models; 3) The metastatic tumor model after the experiment is mounted on a stereotaxic assembly and scanned using an image scanning device to obtain a dose-response model of the sparing metastatic tumor treatment effect.

10. The device according to any one of claims 1 to 7 or the method according to claim 8 or 9, characterized in that: The image scanning devices are CT and MR.

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