CN115040282A - Device for constructing brain metastatic cancer animal model and application of device in medicine - Google Patents
Device for constructing brain metastatic cancer animal model and application of device in medicine Download PDFInfo
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Abstract
The invention discloses a device for constructing a brain metastatic cancer animal model and application thereof in medicine, the invention constructs a mouse model of the internal carotid artery injection brain metastatic cancer which is more in line with the normal physiological characteristics of a human body for the first time, compared with the brain metastatic cancer animal model reported in the prior art, the animal model of the brain metastatic cancer which is constructed by the device in the invention is more in line with the normal physiological characteristics of the human body, provides a brand-new animal model of the brain metastatic cancer of a non-human mammal for the field, can be used for researching the pathogenesis of the brain metastatic cancer, and can be used for screening new drugs and testing effectiveness.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and relates to a device for constructing an animal model, in particular to a device for constructing a brain metastatic cancer animal model and application thereof in medicines.
Background
With the increasing progress of industrialization and urbanization, the gradual increase of population aging, and a plurality of risk factors such as chronic infection, unhealthy life style, environmental pollution, cancer has become one of the bigger public problems affecting human survival all over the world today, and the morbidity and mortality of cancer are increasing year by year (Cao Mao, Chen Wan. GLOBOCAN 2020 global cancer statistics interpretation [ J ]. China advanced journal of medicine (electronic edition), 2021,13(03): 63-69.). With the widespread application of nuclear magnetic resonance technology in clinical diagnosis, the blood Brain barrier function of various novel chemotherapeutics and targeted drugs may be damaged to different degrees while the drugs permeate the blood Brain barrier, the curative effect of malignant tumor treatment is improved, the incidence of Brain metastatic cancers (BM) is further increased, and about 20% -40% of cancer patients in the course of the disease have Brain metastatic cancers (Tsao MN, Lloyd N, Wong RK, Chow E, Rakovitch E, laperie N, Xu W, Sahgal a. white Brain metabolic therapies for the treatment of new diagnogged multiple chemotherapy, cochrane Database replacement v.2012apr 18; 2012(4): CD 003869).
Brain metastasis refers to metastatic tumor in which extracranial malignant tumor affects brain, meninges, intracerebral vessels and cranial nerves, the incidence rate of the metastatic cancer far exceeds that of primary malignant tumor of brain, the metastasis to parenchyma of brain is most common, and the metastasis to meninges is the second. Brain metastasis of malignant tumor is one of the common tumor complications, usually tumor cells fall from a primary focus and enter into the cranium along with blood flow, and the most common primary tumor is a lung malignant tumor or other malignant tumor which is transferred into the lung and then transferred into the cranium, and then transferred into the spinal cord or the cranium through a vertebral vein. Tumor metastasis is located at the gray-white junction, probably because the blood flow is dominated by different arteries, the blood vessels change in direction, the blood vessels become narrow and the like, so that tumor cells are easy to stay at the junction. The current common treatment modes of the brain metastasis cancer comprise modes of operation, radiotherapy, chemotherapy, intrathecal injection chemotherapy, targeted therapy and the like, but the treatment effect is not optimistic and an effective treatment means is still lacked.
One of the recognized drawbacks in the field of cancer research is the lack of a model system capable of studying carcinogenesis and cancer treatment, and in order to evaluate the efficacy of cancer treatment in preclinical and clinical studies, tumor animal models in which human tumor cell lines or patient-derived tumors are transplanted into immunodeficient mice are commonly used. At present, the blood vessels at the side of the tumor cells injected into the carotid artery injection animal model of brain metastatic cancer reported in related researches are ligated, which results in the blocking of blood supply into the skull, the compensation capacity of general experimental animals such as mice is stronger, and no obvious cerebral infarction phenomenon can occur, while in clinical patients, the carotid artery at one side is blocked, which can cause serious clinical symptoms such as insufficient blood supply to the brain and even cerebral infarction, therefore, the carotid artery injection animal model of brain metastatic cancer reported at present does not accord with the normal physiological characteristics of human body.
In order to solve the technical problems faced by the present field, the invention provides a device for constructing a brain metastatic cancer animal model according with the normal physiological characteristics of a human body and the application thereof in medicine.
Disclosure of Invention
In view of the above, the present invention aims to provide a device for constructing an animal model of brain metastatic cancer and a use thereof in medicine, wherein tumor cells are injected into the cranium from the external carotid artery to the internal carotid artery via the common carotid artery by a simple injection device, and finally the external carotid artery is ligated, wherein the internal carotid artery and the common carotid artery are both slipknots and are loosened after injection without blocking the blood entering the cranium at the injection side, so that the animal model of brain metastatic cancer constructed by the device in the present invention more conforms to the normal physiological characteristics of the human body compared with the animal model of brain metastatic cancer reported in the present field.
The above object of the present invention is achieved by the following technical solutions:
the invention provides a device for constructing a brain metastatic cancer animal model.
Further, the device comprises the following units: the device comprises a processing unit, an injection unit and a reset unit;
the processing unit executes the following operation steps:
(1) exposing the right common carotid artery, carotid bifurcation, a portion of the external carotid artery and internal carotid artery of the animal;
(2) respectively knotting exposed proximal end of common carotid artery, external carotid artery near carotid bifurcation, external carotid artery far end and internal carotid artery near carotid bifurcation;
(3) tightening the slipknot at the proximal end of the common carotid artery;
(4) tightening the slipknot of the internal carotid artery near the carotid bifurcation;
(5) tightening and ligating the slipknot at the distal end of the external carotid artery;
the injection unit performs the following operational steps:
(1) the external carotid artery blood vessel is cut off between the external cervical distal end and the proximal end;
(2) injecting tumor cells into the internal carotid artery;
(3) tightening the knot of the external carotid artery near the carotid bifurcation;
(4) loosening the proximal end knot of the common carotid artery;
(5) loosening the knot of the internal carotid artery close to the carotid bifurcation;
the reset unit executes the following operation steps:
and (4) resetting the separated muscle, and sealing the skin to obtain the brain metastasis cancer animal model.
Further, the animal is a non-human mammal;
preferably, the non-human mammal includes a mouse, rat, guinea pig, rabbit, monkey, sheep, goat;
more preferably, the non-human mammal is a mouse;
most preferably, the mice include tumor cell line syngeneic mice, syngeneic transgenic mice, immunodeficient mice;
most preferably, the mouse is an immunodeficient mouse;
most preferably, the mouse is a BALB/c mouse;
most preferably, the BALB/c mouse has a mass of 18-22 g.
In actual experimental procedures, the corresponding germ-line mice can be selected according to the experimental purpose, for example: (1) carrying out immunity related experiments, wherein tumor cell line homologous mice or homologous transgenic mice can be selected; (2) when a human tumor model is established, an immunodeficiency mouse is selected, the specification of the mouse germ line is not intended to limit the protection scope of the invention, but is a further supplementary specification, as long as an animal model constructed by using the device disclosed by the invention falls into the protection scope of the invention, in the embodiment of the invention, an 8 +/-2 week immunodeficiency mouse BALB/c female mouse is selected.
Further, the step (1) of exposing the right common carotid artery, the bifurcation of the carotid artery, part of the external carotid artery and the internal carotid artery of the animal further comprises the following steps: removing hair from the neck of the animal and cleansing the exposed neck skin;
preferably, iodine and/or 75% alcohol is used to cleanse the skin.
Further, the step (1) of exposing the right common carotid artery, the bifurcation of the carotid artery, part of the external carotid artery and the internal carotid artery of the animal further comprises the following steps: making an incision in the cleaned neck skin;
preferably, the incision is obtained by cutting the cleaned neck skin by using a surgical scalpel;
preferably, the incision is about 1 cm.
Further, the tool for exposing the right common carotid artery, carotid bifurcation, part of external carotid artery and internal carotid artery of the animal in the step (1) is preferably blunt-cutting muscle forceps.
Further, after exposing the right common carotid artery, the bifurcation of the carotid artery, a part of the external carotid artery and the internal carotid artery of the animal as described in the step (1), a wetted cotton ball was placed under the carotid artery at the injection site.
Further, the tightening of the proximal kink of the common carotid artery in step (3) serves to stop the flow of blood from the heart into the injection area.
Further, the tightening of the kink in the internal carotid artery near the carotid bifurcation in step (4) serves to prevent the backflow of blood from the intracranial space, affecting the injection and tumorigenic effects.
Further, the effect of tightening and ligating the slip knot at the distal end of the external carotid artery as described in step (5) is to block the backflow of blood from outside the neck.
Further, the step (7) of injecting the tumor cells into the common carotid artery is performed by using a simple injection device.
Further, the simple injection device is formed by inserting a needle of a syringe into the display tube;
preferably, the needle of the syringe is 30G or 31G;
more preferably, the needle of the syringe is 30G;
preferably, the microtube is preferably PE 10.
Further, the step of injecting tumor cells into the common carotid artery as described in step (7) further comprises the steps of: the distal end display micro-tube of the simple injection device is inserted into a blood vessel through the broken end of the external carotid artery close to the bifurcation of the carotid artery, after the distal end display micro-tube passes through the bifurcation of the carotid artery to reach the common carotid artery, the junction of the internal carotid artery close to the bifurcation of the carotid artery is properly loosened, so that the distal end of the micro-tube enters the internal carotid artery again, and after crossing the junction of the internal carotid artery, the junction is properly tightened and changed, so that the distal end of the micro-tube is fixed in the internal carotid artery blood vessel.
Further, the tumor cells in step (7) include breast cancer cells, lung cancer cells, choriocarcinoma cells, melanoma cells, kidney cancer cells, nasopharyngeal cancer cells, cancer cells of the mouth floor, retinoblastoma cells, tumor cells of the digestive system;
preferably, the tumor cells comprise 4T1 murine breast cancer cells, MDA-MB-231A human breast cancer cells, human lung cancer cells;
more preferably, the tumor cell is a 4T1 murine breast cancer cell;
most preferably, the tumor cells are used in an amount of 2X 10 5 ～5×10 5 cells/mL。
In actual experimental operation, the injected tumor cell type and the amount of tumor cells can be adjusted according to experimental purposes and experimental needs, and the adjustment also falls into the protection scope of the present invention.
Furthermore, the operation of the steps adopts an aseptic technique.
Further, tightening the external carotid artery knot near the carotid bifurcation in step (8) further comprises the following steps: after the injection process is finished, the PE display micro-tube is pulled out, and the external carotid artery is tightened and tied to a knot close to the carotid bifurcation.
Further, the act of loosening the proximal carotid node in step (9) is to allow blood from the heart to fill the injection area.
Furthermore, after the internal carotid artery is loosened and the knot is closed to the bifurcation of the carotid artery in the step (10), blood smoothly enters the intracranial through the internal carotid artery due to the pressure effect, thereby not only avoiding the backflow of tumor cells entering the intracranial, but also not blocking important blood vessels in the intracranial and maintaining the normal physiological state.
In a second aspect the invention provides a cell or cell line or primary cell culture or tissue or organ or culture thereof.
Further, the cell or cell line or primary cell culture or tissue or organ or culture thereof is derived from an animal model of brain metastatic cancer or progeny thereof constructed by the device of the first aspect of the invention.
In a third aspect, the invention provides the use of a cell or cell line or primary cell culture or tissue or organ or culture thereof according to the second aspect of the invention for the non-diagnostic and non-therapeutic screening of candidate agents for the prevention and/or treatment of brain metastases.
In a fourth aspect, the invention provides the use of a cell or cell line or primary cell culture or tissue or organ or culture thereof according to the second aspect of the invention for the non-diagnostic and non-therapeutic evaluation of the effectiveness of a candidate drug for the prevention and/or treatment of brain metastases.
In a fifth aspect of the invention, a method for non-diagnostically and non-therapeutically screening a candidate drug for the prevention and/or treatment of brain metastasis is provided.
Further, the method comprises the steps of:
(1) administering a test substance to a cell or cell line or primary cell culture or tissue or organ or culture thereof according to the second aspect of the invention, said method comprising a test substance treatment group and a control group;
(2) observing the migration of the cells or cell lines or primary cell cultures of the second aspect of the invention or the growth of the tissue or organ or culture thereof of the second aspect of the invention in step (1);
(3) if migration of the cell or cell line or primary cell culture according to the second aspect of the invention is inhibited, or growth of the tissue or organ or culture thereof according to the second aspect of the invention is inhibited, the test agent is a candidate drug.
A sixth aspect of the present invention provides a method for non-diagnostically and non-therapeutically evaluating the effectiveness of a candidate drug for the prevention and/or treatment of brain metastatic cancer.
Further, the method comprises the steps of:
(1) administering a candidate drug to a cell or cell line or primary cell culture or tissue or organ or culture thereof according to the second aspect of the invention, said method comprising treating a test agent with a test agent and a control agent;
(2) observing the migration of the cells or cell lines or primary cell cultures of the second aspect of the invention or the growth of the tissue or organ or culture thereof of the second aspect of the invention in step (1);
(3) if the inhibition of migration of the cell or cell line or primary cell culture according to the second aspect of the invention, or growth of the tissue or organ or culture thereof according to the second aspect of the invention is inhibited by more than 50%, this indicates that the candidate drug is effective for the prevention and/or treatment of brain metastatic cancer.
The invention has the advantages and beneficial effects that:
the invention constructs an internal carotid artery injection brain metastatic cancer mouse model which is more accordant with the normal physiological characteristics of a human body for the first time, tumor cells are injected into the skull from an external carotid artery to an internal carotid artery through a simple injection device, and finally the external carotid artery is ligated, and the internal carotid artery are all slipknots and loosened after injection without blocking intracranial blood on one side of injection.
Drawings
Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:
FIG. 1 is a schematic diagram showing the construction of an animal model for the injection of brain metastasis via the external carotid artery and the internal carotid artery of an external carotid artery;
fig. 2 is a graph showing the result of intracranial tumorigenesis of an animal model of brain metastatic cancer constructed according to the present invention, in which arrows indicate tumor regions, a graph: layer 1, panel B: layer 2, panel C: layer 3, D diagram: layer 4, panel E: layer 5, panel F: and (6) layer.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and are not to be construed as limiting the invention. As will be understood by those of ordinary skill in the art: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents. The following examples are examples of experimental methods not indicating specific conditions, and the detection is usually carried out according to conventional conditions or according to the conditions recommended by the manufacturers.
Example 1 construction of mouse internal carotid artery injection brain metastasis cancer model
1. Experimental Material
The mouse is a BALB/c mouse which is purchased from Beijing Wintonlihua laboratory animal technology GmbH, and the tumor cell is a 4T1 murine breast cancer tumor cell which is purchased from the molecular cell science and innovation center of Chinese academy of sciences.
2. Experimental method
In this embodiment, all surgical operations are performed by aseptic technique, and the detailed construction process is as follows:
(1) after fixing BALB/c mice, removing the hair on the neck of the mice by using scissors, and cleaning the skin on the neck of the mice by using iodine and 75% alcohol;
(2) an incision of about 1 cm in length was made in the bare skin of the mouse with a surgical scalpel;
(3) the right common carotid artery, carotid bifurcation, part of external carotid artery and internal carotid artery of the mouse are exposed by blunt-cutting muscle forceps;
(4) placing a small wet cotton ball under the carotid artery of the injection site to wet and protect the blood vessel;
(5) respectively knotting the exposed proximal end of the common carotid artery, the position of the external carotid artery close to the bifurcation of the carotid artery, the distal end of the external carotid artery and the position of the internal carotid artery close to the bifurcation of the carotid artery (see figure 1A);
(6) A30G needle of a 1mL syringe containing 4T1 tumor cells was inserted into a microtube (PE10) to form a simple injection device, and the distal end of the PE microtube was ready for use.
(7) Tightening the kink at the proximal end of the common carotid artery to stop blood flow from the heart into the injection zone;
(8) tightening the slipknot of the internal carotid artery near the carotid bifurcation to prevent blood backflow from the intracranial and affect the injection and tumorigenic effect;
(9) tightening and ligating the slipknot at the distal end of the external carotid artery to block the backflow of blood from outside the neck;
(10) the external carotid artery blood vessel is cut off between the external cervical distal end and the proximal end;
(11) inserting a distal PE (polyethylene) microtube of the simple injection device into a blood vessel through the broken end of an external carotid artery close to the bifurcation of a carotid artery, properly loosening a knot at the position of an internal carotid artery close to the bifurcation of the carotid artery after the PE microtube is bifurcated into a common carotid artery, enabling the distal end of the PE microtube to enter the internal carotid artery again, crossing the knot of the internal carotid artery, and properly tightening the loose knot (see figure 1B, C), namely fixing the distal end of the PE microtube in the blood vessel of the internal carotid artery, and preventing the backflow of intracranial distal blood;
(12) slowly injecting the tumor cells in the syringe into the internal carotid artery;
(13) after the injection is finished, rapidly tightening knots of the external carotid artery close to the bifurcation of the carotid artery, completing the injection process, pulling out the PE display micro-tube, tightening and ligating the knots of the external carotid artery close to the bifurcation of the carotid artery, and trimming redundant silk threads by using micro-scissors;
(14) loosening the proximal node of the common carotid artery to allow blood from the heart to fill the injection area;
(15) the node of the internal carotid artery close to the bifurcation of the carotid artery is loosened, and the blood smoothly enters the intracranial space through the internal carotid artery under the action of pressure, thereby not only avoiding the backflow of tumor cells entering the intracranial space, but also not blocking important blood vessels in the intracranial space, and maintaining the normal physiological state of the intracranial blood supply (see figure 1D);
(16) repositioning the isolated muscle to cover the wound site;
(17) the skin is closed with nylon or skin adhesive.
3. Results of the experiment
The schematic diagram of the animal model of the brain metastasis tumor injected through the internal carotid artery during the external carotid artery is shown in figures 1A-D, the experimental result shows that after the model is established, the mouse is observed every day, after 2-3 weeks, the mouse is killed, the brain tissue of the mouse is obtained and is sliced, as shown in figures 2A-F, the tumor conditions of different layers of the brain tissue of the mouse of the same model are shown, 4T1 tumor cells are marked by Lucifer red fluorescence, the brain tissue is subjected to DAPI staining, the arrow is shown as a tumor area, the experimental result further shows the successful construction of the animal model of the brain metastasis tumor, and the obtained mouse model is more in line with the normal physiological characteristics of a human body because the construction method does not block intracranial blood at the injection side.
Embodiment 2 an apparatus for constructing animal model of brain metastatic cancer
The application provides a device for constructing a brain metastatic cancer animal model, which comprises the following units: the device comprises a processing unit, an injection unit and a reset unit;
the processing unit executes the following operation steps:
(1) exposing the right common carotid artery, carotid bifurcation, part of the external carotid artery and the internal carotid artery of BALB/c mice with immunodeficient mice;
(2) respectively knotting exposed proximal end of common carotid artery, external carotid artery near carotid bifurcation, external carotid artery far end and internal carotid artery near carotid bifurcation;
(3) tightening the slipknot at the proximal end of the common carotid artery;
(4) tightening the slipknot of the internal carotid artery near the carotid bifurcation;
(5) tightening and ligating the slipknot at the distal end of the external carotid artery;
the injection unit performs the following operational steps:
(1) the external carotid artery blood vessel is cut off between the external cervical distal end and the proximal end;
(2) 2 x 10 to 5 ～5×10 5 cells/mL 4T1 murine breast cancer cells are injected into the internal carotid artery, a simple injection device is adopted for injecting the tumor cells into the common carotid artery, the simple injection device is formed by inserting a 30G needle of a syringe into a PE10 microscopic tube, and the concrete steps are as follows: inserting a distal microscopic tube of the simple injection device into a blood vessel through the broken end of an external carotid artery close to the bifurcation of a carotid artery, properly loosening a knot at the position of an internal carotid artery close to the bifurcation of the carotid artery after the distal microscopic tube passes through the bifurcation of the carotid artery and reaches a common carotid artery, and enabling the distal end of the microscopic tube to enter the internal carotid artery again and cross the knot of the internal carotid artery;
(3) tightening the knot of the external carotid artery near the carotid bifurcation;
(4) loosening the proximal end knot of the common carotid artery;
(5) loosening the knot of the internal carotid artery close to the carotid bifurcation;
the reset unit executes the following operation steps:
and (4) resetting the separated muscle, and sealing the skin to obtain the brain metastasis cancer animal model.
Example 3 verification of mouse model of internal carotid artery injection brain metastasis carcinoma constructed by the construction method or the construction device of the invention
In the application, the internal carotid artery injection brain metastasis cancer mouse model constructed by the construction method described in embodiment 1 or the construction device described in embodiment 2 is adopted, and experiments show that under the condition of being constructed strictly according to the construction method and the construction device described in the invention, the success rate of model construction is very high, none of the constructed mouse models fails, and the constructed internal carotid artery injection brain metastasis cancer mouse model is more in line with the normal physiological characteristics of a human body compared with the brain metastasis cancer mouse model constructed by the traditional method and the brain metastasis cancer mouse model disclosed in the related documents or reports at present.
The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.
Claims (10)
1. An apparatus for constructing an animal model of brain metastatic cancer, wherein the apparatus comprises the following units: the device comprises a processing unit, an injection unit and a reset unit;
the processing unit executes the following operation steps:
(1) exposing the right common carotid artery, carotid bifurcation, part of the external carotid artery and internal carotid artery of the animal;
(2) respectively knotting exposed proximal end of common carotid artery, external carotid artery near carotid bifurcation, external carotid artery far end and internal carotid artery near carotid bifurcation;
(3) tightening the slipknot at the proximal end of the common carotid artery;
(4) tightening the slipknot of the internal carotid artery near the carotid bifurcation;
(5) tightening and ligating the slip knot at the distal end of the external carotid artery;
the injection unit performs the following operational steps:
(1) the external carotid artery blood vessel is cut off between the external cervical distal end and the proximal end;
(2) injecting tumor cells into the internal carotid artery;
(3) tightening the knot of the external carotid artery close to the bifurcation of the carotid artery;
(4) loosening the proximal end knot of the common carotid artery;
(5) loosening the knot of the internal carotid artery close to the carotid bifurcation;
the reset unit executes the following operation steps:
and (4) resetting the separated muscle, and sealing the skin to obtain the brain metastasis cancer animal model.
2. The device of claim 1, wherein the animal is a mouse;
preferably, the mouse is an immunodeficient mouse;
more preferably, the immunodeficient mouse is a BALB/c mouse;
most preferably, the BALB/c mouse has a mass of 18-22 g.
3. The device of claim 1, wherein the injection of the tumor cells into the common carotid artery in step (2) of the injection unit is performed using a simple injection device;
preferably, the simple injection device is formed by inserting a needle of a syringe into the microtube;
more preferably, the needle of the syringe is 30G or 31G;
most preferably, the needle of the syringe is 30G;
most preferably, the microtube is preferably PE 10.
4. The device of claim 1, wherein the injecting the tumor cells into the common carotid artery in step (2) of the injection unit further comprises the steps of: the distal microscopic tube of the simple injection device of claim 3 is inserted into the blood vessel through the broken end of the external carotid artery near the bifurcation of the carotid artery, and after the distal microscopic tube passes through the bifurcation of the carotid artery to the common carotid artery, the knot of the internal carotid artery near the bifurcation of the carotid artery is properly released, so that the distal end of the microscopic tube enters the internal carotid artery again and crosses the knot of the internal carotid artery.
5. The apparatus according to claim 4, wherein the tumor cells in step (2) of the injection unit comprise breast cancer cells, lung cancer cells, choriocarcinoma cells, melanoma cells, kidney cancer cells, nasopharyngeal cancer cells, cancer cells of the mouth bottom, retinoblastoma cells, tumor cells of the digestive system;
preferably, the tumor cells comprise 4T1 murine breast cancer cells, MDA-MB-231A human breast cancer cells, human lung cancer cells;
more preferably, the tumor cell is a 4T1 murine breast cancer cell;
most preferably, the tumor cells are used in an amount of 2X 10 5 ～5×10 5 cells/mL。
6. A cell or cell line or primary cell culture or tissue or organ or culture thereof derived from an animal model of brain metastasis cancer or the progeny thereof constructed using the device of any one of claims 1-5.
7. Use of the cell or cell line or primary cell culture or tissue or organ or culture thereof according to claim 6 for the non-diagnostic and non-therapeutic screening of candidate agents for the prevention and/or treatment of brain metastases.
8. Use of the cell or cell line or primary cell culture or tissue or organ or culture thereof of claim 6 for the non-diagnostic and non-therapeutic evaluation of the effectiveness of a candidate drug for the prevention and/or treatment of brain metastatic cancer.
9. A method for non-diagnostic and non-therapeutic screening of candidate drugs for the prevention and/or treatment of brain metastasis, comprising the steps of:
(1) administering a test substance to the cell or cell line or primary cell culture or tissue or organ or culture thereof of claim 6, said method comprising a test substance treatment group and a control group;
(2) observing the migration of the cell or cell line or primary cell culture of claim 6 or the growth of the tissue or organ or culture thereof of claim 6 in step (1);
(3) if the migration of the cell or cell line or primary cell culture of claim 6 is inhibited, or the growth of the tissue or organ or culture thereof of claim 6 is inhibited, the test substance is a drug candidate.
10. A method for non-diagnostically and non-therapeutically evaluating the effectiveness of a candidate drug for the prevention and/or treatment of brain metastatic cancer, the method comprising the steps of:
(1) administering a candidate drug to the cell or cell line or primary cell culture or tissue or organ or culture thereof of claim 6, said method comprising a test agent treatment group and a control group;
(2) observing the migration of the cell or cell line or primary cell culture of claim 6 or the growth of the tissue or organ or culture thereof of claim 6 in step (1);
(3) if the inhibition of migration of the cell or cell line or primary cell culture of claim 6, or growth of the tissue or organ or culture thereof of claim 6 is greater than 50%, it is indicative that the candidate drug is effective for the prevention and/or treatment of brain metastatic cancer.
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| CA2497818C (en) * | 2004-03-19 | 2010-10-12 | F. Hoffmann-La Roche Ag | Glyt1 transgenic mice |
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| CN105101903A (en) * | 2013-02-04 | 2015-11-25 | 儿童国家医疗中心 | Hybrid control surgical robotic system |
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| CN107019559A (en) * | 2016-11-16 | 2017-08-08 | 温州医科大学附属眼视光医院 | Full-automatic ophthalmologic operation robot |
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