CN110623770A

CN110623770A - Method for constructing malignant schwannoma xenograft mouse model

Info

Publication number: CN110623770A
Application number: CN201910900585.5A
Authority: CN
Inventors: 黎志宏; 万璐; 刘忠越; 段智曦; 张程豪; 罗煦君
Original assignee: Second Xiangya Hospital of Central South University
Current assignee: Second Xiangya Hospital of Central South University
Priority date: 2019-09-23
Filing date: 2019-09-23
Publication date: 2019-12-31
Anticipated expiration: 2039-09-23
Also published as: CN110623770B

Abstract

The invention provides a method for constructing a malignant schwannoma xenograft mouse model, which comprises the steps of recovering a malignant schwannoma patient tumor tissue block stored in a liquid nitrogen environment, then transplanting the recovered patient tumor tissue block into an immunodeficiency mouse under an aseptic condition in a heterogenic subcutaneous way, wherein the heterogenic subcutaneous transplantation comprises the steps of selecting an immunodeficiency mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with the sex of the patient, soaking the recovered patient tumor tissue block into a sterile incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, implanting the patient tumor tissue block into the mouse under the skin, suturing an incision, and feeding the mouse to enable the tumor in the mouse to grow. The invention also comprises the step of transplanting the tumor tissue blocks of the cryopreserved resuscitated patients into immunodeficient mice in a heterogenic and in-situ manner. The invention uses the xenograft mouse to replace clinical patients to screen different combined drugs, thereby selecting the most suitable drug, greatly improving the effectiveness and reducing the side effect of the drug.

Description

Method for constructing malignant schwannoma xenograft mouse model

Technical Field

The invention relates to the field of establishment of tumor transplantation mouse models, in particular to a method for establishing a malignant schwannoma xenograft mouse model.

Background

Malignant schwannoma, also known as neurosarcoma, is derived from the peripheral nerve sheath, consists of malignant neuroblasts, and is mostly malignant from neurofibroma or schwannoma. The malignant degree is high, lung metastasis is easy to occur, the disability rate and the death rate are high, and the prognosis is poor. Malignant schwannoma imposes a heavy economic burden on society and even countries, and brings about a greater social problem. Because of the heterogeneity of malignant schwannoma, the basic research results based on malignant schwannoma cell line in the past can be rarely reproduced in patients with clinical malignant schwannoma, and the low disease incidence results in the delay of the diagnosis and treatment level of clinical malignant schwannoma.

The invention patent application CN201710822683.2 discloses human NF2^-/-A method for establishing a schwann cell line of vestibular nerve sheath tumor and a cell line thereof. The establishing method comprises the following steps: A. separating Schwann cells in tumor to obtain human NF2^-/-Performing primary culture on the primary Schwann cells; B. establishment of immunodeficient sciatic nerve human NF2 in mouse^-/-Primary Schwann cell transplantation tumor model; C. screening transplanted tumors with the diameter of more than 1cm from sciatic nerves of mice; D. separating Schwann cells, and performing primary cell culture; E. carrying out continuous subculture for more than 5 generations; F. obtained by screening with human NF2^-/-The cell morphology and the main biological characteristics of the primary Schwann cells and the primary transplantation tumor cell strains are consistent, and the NF2 can be subjected to unlimited passage^-/-Vestibular nerve Schwann cell.

Tumor models are important vehicles for tumor research. Researches show that the cells are continuously adapted to in-vitro environment in the passage process, the biological characteristics are not in accordance with those of in-vivo tumor cells, and the cell line xenograft model has greater consistency with the tumor of a patient, so that the failure rate of clinical tests of the antitumor drugs is high.

A human derived tumor xenograft (PDX) model is an in-vivo model, and a large amount of tumor tissues of a patient can be amplified by taking a mouse as a carrier by directly transplanting fresh tumor tissues of the patient to an immunodeficiency mouse after being treated and depending on the environment growth and gradual passage provided by the mouse. The PDX model has the following advantages: firstly, the model is equivalent to a model corresponding to a patient, can well reflect the difference of tumors among different individuals, and can better reflect the actual condition of clinical patients compared with the traditional model; secondly, the PDX model can be passaged, and the passaged tumor tissue can keep high consistency with the initial tumor tissue in gene copy number, gene mutation, expression mode and the like, which is equivalent to the amplification of a patient tumor sample, thereby providing good conditions for drug screening and the study of a patient disease mechanism; and thirdly, the tumor blocks cultured by the PDX can be frozen for resuscitation, and a new PDX model can still be constructed, which is equivalent to the construction of a tumor living body library of different patients.

The existing technology is mainly to perform subcutaneous modeling after fresh malignant nerve sheath tumor tissues are subjected to a chopping treatment or a cell homogenization treatment.

Disclosure of Invention

However, homogenization of fresh malignant schwannoma tissue is considered to destroy the microenvironment of tumor tissue itself, and the consistency of tumor and surrounding stroma with patients cannot be well maintained. In addition, malignant schwannoma is derived from the nerve sheath, and compared with subcutaneous modeling, in-situ modeling can better simulate the tumorigenic environment. The method successfully establishes a tissue block transplantation mode, well maintains the stable relation between the tumor and the peripheral interstitium, successfully establishes subcutaneous modeling and nerve sheath in-situ modeling of malignant nerve sheath tumor, and maintains the consistency of the growth environment of the model tumor and the tumor of a patient.

Therefore, the invention firstly provides a method for constructing a malignant schwannoma xenograft mouse model, which comprises the steps of collecting a malignant schwannoma patient tumor tissue block, carrying out xenograft on the fresh patient tumor tissue block into an immunodeficiency mouse body under aseptic conditions, wherein the xenograft comprises selecting an immunodeficiency mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with the sex of the patient, soaking the collected fresh patient tumor tissue block into an aseptic incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, implanting the patient tumor tissue block into the mouse under the skin, suturing an incision, and feeding the mouse to ensure the tumor in the mouse to grow.

The invention also provides a method for constructing a malignant schwannoma xenograft mouse model, which comprises the steps of collecting a malignant schwannoma patient tumor tissue block, transplanting a fresh patient tumor tissue block into an immunodeficiency mouse in a xenograft in situ under aseptic conditions, wherein the xenograft in situ comprises selecting an immunodeficiency mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with the sex of the patient, soaking the collected fresh patient tumor tissue block in a sterile incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, implanting the patient tumor tissue block into a nerve sheath of the mouse, suturing an incision, and feeding the mouse to enable the tumor in the mouse to grow.

In a specific embodiment, the subcutaneous implantation comprises making a skin incision in the mouse axilla, implanting tumor tissue subcutaneously, and suturing the surgical incision; the orthotopic transplantation comprises making a skin incision at the axillary nerve sheath or sciatic nerve sheath of the mouse, separating the nerve sheath and opening, implanting a tumor tissue block of the patient into the nerve sheath, and then suturing the surgical incision.

In a specific embodiment, the tumor tissue mass of the malignant schwannoma patient implanted in the mouse has a unilateral length of 3mm or less, preferably 1mm or less.

In a specific embodiment, the weight of the mouse in subcutaneous transplantation or orthotopic transplantation is 8-20 g, preferably 12-15g, the sterile incubation liquid further comprises an antibiotic, preferably the sterile incubation liquid comprises an antibiotic and a DMEM medium or an RPMI-1640 medium, and more preferably the antibiotic comprises penicillin and streptomycin.

In a specific embodiment, the immunodeficient mouse is a NOD/SCID mouse.

In a specific embodiment, the method further comprises observing the size of the malignant schwannoma in the mouse using a small animal living body imager, preferably also using the small animal living body imager to observe whether the tumor has metastasized to the internal organs of the mouse.

In a specific embodiment, the method further comprises raising the mouse such that the mouse is dissected after the growth of the tumor in vivo, and the tumor is detached to obtain a tumor mass for P0 generation, and the tumor mass for P0 generation is taken as a tumor mass for xenogeneic subcutaneous transplantation or xenogeneic orthotopic transplantation into the immunodeficient mouse to obtain a tumor mass for P1 generation, and optionally the tumor mass for P1 generation is taken as a tumor mass for xenogeneic subcutaneous transplantation or xenogeneic orthotopic transplantation into the immunodeficient mouse to obtain a tumor mass for P2 generation, and so on for passage; preferably the method further comprises the step of programming the P0 generation tumor mass to be cooled and stored in liquid nitrogen, and optionally the steps of programming the P1 and P2 generation tumor mass to be cooled and stored in liquid nitrogen.

The invention also provides a construction method of the mouse model for the heterotransplantation of malignant schwannoma, which comprises the steps of heterogeneously and subcutaneously transplanting or heterogeneously and in situ transplanting P0 generation malignant schwannoma tumor tissues, P1 generation malignant schwannoma tumor tissues or P2 generation malignant schwannoma tumor tissues in a fresh mouse model into an immunodeficient mouse under the aseptic condition, wherein the heterogeneously and subcutaneously transplanting or heterogeneously and in situ transplanting comprises selecting the immunodeficient mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with that of a patient, soaking the malignant schwanoma tumor tissue block into sterile incubation liquid containing fetal calf serum, anesthetizing the mouse, fixing, implanting the malignant schwanoma tumor tissue block into the subcutaneous or nerve sheath of the mouse, suturing the incision, and feeding the mouse to ensure the growth of tumors in the body.

The invention also provides application of the mouse model obtained by the method in screening drugs for preventing or treating malignant schwannoma.

The invention also provides a method for constructing a malignant schwannoma xenograft mouse model, which is characterized by comprising the steps of recovering a malignant schwannoma patient tumor tissue block stored in a liquid nitrogen environment, transplanting the recovered malignant schwannoma patient tumor tissue block into an immunodeficiency mouse under an aseptic condition in a heterogenic subcutaneous way, wherein the heterogenic subcutaneous transplantation comprises the steps of selecting an immunodeficiency mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with the sex of the patient, soaking the recovered patient tumor tissue block into sterile incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, implanting the patient tumor tissue block into the subcutaneous way of the mouse, suturing an incision, and feeding the mouse to enable the tumor in the mouse to grow.

The invention also provides a method for constructing a malignant schwannoma xenograft mouse model, which is characterized by comprising the steps of recovering a malignant schwannoma patient tumor tissue block stored in a liquid nitrogen environment, transplanting the recovered malignant schwannoma patient tumor tissue block into an immunodeficient mouse in a heterogenic and in-situ manner under an aseptic condition, wherein the heterogenic and in-situ transplantation comprises the steps of selecting an immunodeficient mouse with the age of 2-6 weeks, keeping the sex of the mouse consistent with the sex of the patient, soaking the recovered malignant schwannoma tissue block of the patient in a sterile incubation liquid containing fetal calf serum, anesthetizing the mouse, fixing, implanting the tumor tissue block of the patient into a nerve sheath of the mouse, suturing an incision, and feeding the mouse to ensure the growth of the tumor in the body.

In a specific embodiment, the method further comprises the processes of programmed cooling and liquid nitrogen preservation and rapid heating resuscitation of the collected tumor tissue mass of the malignant schwannoma patient.

In a specific embodiment, the tumor tissue block of the patient with malignant schwannoma is preserved in DMEM containing DMSO, FBS and NEAA, more than three sections of programs are cooled to the temperature of liquid nitrogen and preserved in the temperature of liquid nitrogen, and the rapid heating resuscitation refers to resuscitating frozen tissue in a constant-temperature water bath at 36-38 ℃.

In a specific embodiment, the immunodeficient mouse is a NOD/SCID mouse, and the unilateral length of the tumor tissue mass of the malignant schwannoma patient implanted in the mouse is less than or equal to 3mm, preferably less than or equal to 1 mm.

In a specific embodiment, the method further comprises observing the size of the malignant schwannoma in the mouse by using a small animal living body imager, and preferably observing whether the tumor is transferred to the internal organs of the mouse by using the small animal living body imager.

The invention also provides a method for constructing a mouse model of malignant schwannoma xenograft, which comprises the steps of carrying out xenogeneic subcutaneous transplantation or xenogeneic in situ transplantation on P0 generation malignant schwannoma tumor tissues, P1 generation malignant schwannoma tumor tissues or P2 generation malignant schwannoma tumor tissues in a mouse model which is frozen in a liquid nitrogen environment and revived under an aseptic condition, soaking the malignant schwannoma tumor tissue blocks in sterile incubation liquid containing fetal calf serum, anaesthetizing the mice and then fixing, implanting the malignant schwannoma tumor tissue blocks into the subcutaneous tissues or the nerve sheath of the mice, suturing an incision, and feeding the mice to enable the tumors to grow in vivo.

The feeding of the mice is specifically feeding the mice in an SPF-level environment.

The invention has at least the following beneficial effects:

the invention utilizes the PDX model with better homogeneity with the primary tumor of the patient, can deeply discuss the pathogenesis of malignant schwannoma, can be well repeated in the body of the patient based on the result developed by PDX, can greatly expand limited malignant schwannoma samples in the body of the clinical patient, and uses a xenograft mouse to replace the clinical patient to carry out different combined drug screening, thereby selecting the most suitable drug, greatly improving the effectiveness and reducing the side effect of the drug.

The PDX model establishment and application of the invention can hopefully improve the effectiveness of clinical malignant nerve sheath tumor treatment and the prognosis of patients, prevent the patients from disability and even death to bring great medical social burden, reduce treatment cost and government financial expenditure, have great practical significance in these aspects, and have great economic benefits.

In addition, the research on malignant nerve sheath tumor is lagged, the animal model which can be used for research per se is few, and the difference with the clinic is large, the in-situ implantation PDX model can better reflect the actual situation of a patient, and a better animal model is provided for the basic research and drug screening of malignant nerve sheath tumor. That is, in situ modeling is more realistic than subcutaneous modeling, the screened drug regimen can be better used on the patient, and in situ modeling is theoretically more likely to have tumor metastasis than subcutaneous modeling.

Drawings

Fig. 1 is a photograph of a pathology report for a patient.

FIG. 2 is a photograph of a tumor tissue mass in an immunodeficient mouse with PDX modeling.

FIG. 3 is a photograph of the success of primary cell culture on fresh patient tumor tissue blocks.

Detailed Description

Approved by the ethical committee of hospitals, patients sign an informed consent before sample collection, collected tissues are subjected to histopathological examination, tumor tissue blocks of malignant schwannoma patients are pathologically confirmed, and the malignant schwannoma patients are stored in sterile physiological saline and placed on ice. Dividing the tissue block into 3 parts, and directly treating (see below for details) the tissue block under aseptic conditions in part 1 to perform xenografting into immunodeficient mice; part 2 was placed in DMEM (a medium containing various amino acids and glucose) supplemented with 10% DMSO (dimethyl sulfoxide), 90% FBS (fetal bovine serum) and 1% NEAA (non-essential amino acids), gradient-frozen to-80 ℃, and placed in liquid nitrogen for storage; and the part 3 is put into liquid nitrogen for quick freezing and then is put into the liquid nitrogen for preservation. Wherein, part 1 is used for modeling, part two is used for resuscitation and inoculation, and part three is used for gene detection for submission.

Examples

Subcutaneous inoculation: selecting 10 NOD/SCID mice with 4 weeks of age, namely non-obese diabetic/severe combined immunodeficiency mice, keeping the sex consistent with the sex of a patient (so as to avoid interference of some unknown hormones), weighing about 12-15g, soaking fresh tumor tissue (or tumor tissue of a malignant schwannoma patient frozen and revived by liquid nitrogen) in sterile incubation solution of pure FBS + penicillin and streptomycin (the final concentration of the penicillin is 100U/ml, and the final concentration of the streptomycin is 100U/ml), and cutting into 1 x 1mm small blocks by using sterile scissors. Mice were anesthetized and fixed, skin incisions were made near the axilla on the right side, tumor tissue blocks were implanted subcutaneously (P0), and surgical incisions were sutured. Mice were observed daily for tumor growth. Tumors grow to diameters exceeding 5 mm. Tumor volume was calculated by measuring tumor volume 2 times per week with a vernier caliper and recording tumor length (a) and width (b) (V ═ a × b)²/2). Meanwhile, a small animal living body imager is adopted to observe the viscera transfer condition, and the volume V of the tumor body is increased to 1500mm³When the time is about 42 days later, the animal is dissected, the tumor body is stripped and weighed, part of tumor mass is inoculated and passed, and the rest is placed into liquid nitrogen for preservation.

In-situ nerve sheath implantation: selecting 10 NOD/SCID mice with 4 weeks of age, keeping the sex consistent with the sex of the patient and the body weight of about 12-15g, and collecting fresh tumor tissue (or tumor group preserved by liquid nitrogen and revived)Tissue) in sterile FBS + penicillin and streptomycin (final concentration of penicillin is 100U/ml and final concentration of streptomycin is 100U/ml), cut into 1 x 1mm pieces with sterile scissors, anesthetized, fixed, skin-incised at the axillary or sciatic sheaths, the sheaths are separated and opened, patient tumor tissue pieces are implanted into the sheaths (P0), tension is not too great, and surgical incisions are sutured. The post-operative mice were placed on a 37 ℃ hotbed until resuscitated, and penicillin was routinely administered intraperitoneally to prevent post-operative infection. Mice were observed daily for tumor growth. The skin of the place to be planted is swollen, the size of malignant nerve sheath tumor and the visceral metastasis condition (whether the tumor is transferred to the viscera of a mouse) are observed by using a small animal living body imager every 3 weeks, and the volume of the body to be planted is increased to 1500mm³When the time is about 42 days later, the animal is dissected, the tumor body is stripped and weighed, part of tumor mass is inoculated and passed, and the rest is placed into liquid nitrogen for preservation. The process of preserving tissue mass or whole tumor mass in liquid nitrogen includes programmed cooling, specifically several temperature gradients of 4 ℃, -20 ℃, -80 ℃ (special freezing refrigerator) and liquid nitrogen.

Passage of tumor tissue of malignant schwannoma xenografted mice: inoculating the well-grown P0 generation xenograft to the tumor-forming mouse, and increasing the tumor volume to 1500mm³At the moment or 42 days, the mice are sacrificed, the tumors are completely stripped, the mice are inoculated on the axillary skin, the axillary nerve sheath, the sciatic nerve sheath and the like of 10 NOD/SCID mice (P1) as before, the growth condition of the tumors of the mice is observed every day, the length and the width of the tumors are measured by a vernier caliper as before, the volume of the tumors is calculated, the visceral metastasis condition is observed by a live body imaging instrument of the mice, the animals are dissected after 42 days, the tumors are stripped and weighed, part of the tumors are passaged by analogy (P2, P3 and the like), and the rest of the tumors are cooled in a gradient manner and are placed in liquid nitrogen for storage.

Resuscitating and inoculating primary and passage tumor tissues of malignant nerve sheath tumor: p0, P1 and P2 frozen in liquid nitrogen are taken to replace malignant schwann tumor tissues, quickly re-warmed to normal temperature, and respectively inoculated in the armpit and the nerve sheath of 10 NOD/SCID mice with the same sex, and the method is the same as the method.

Because malignant nerve sheath tumor tissue of a patient is a precious research material, the tumor tissue of tumor-bearing mice needs to be passaged so as to generate more tumor-bearing mice for experiments. The reason why the tumor tissue is frozen and thawed is that too large tumor masses cannot be used in each modeling experiment, and the tumor masses left for seed storage, namely frozen and thawed, can be used in the next experiment. Tumor tissues are passaged through P0, P1, P2 and the like, and the tumor tissues are not changed theoretically, so that the experimental effects of the respective generations can be regarded as the same. And when in-situ modeling or subcutaneous modeling is carried out, as long as the generation of the P0 tumor is successful, subsequent passages and resuscitations of P1, P2 and the like are simple and easy and can be completely successful.

1 patient in this subject group has been collected, and a PDX model was established by in situ and subcutaneous neoplasia to obtain corresponding cases of neoplasia. The patient has recurrence after malignant schwannoma of poplar, male, 28 years old and right thigh. "left popliteal vascular fibromyxoma resection" in 2010, "L3/4 right paraspinal schwannoma resection" in 11 months in 2015, "right femoral schwannoma resection" in 2 months in 2017, "right femoral malignant schwanoma resection" in 12 months in 2017, and "right hip amputation" in 4 months in 2018.

Fig. 1 is a photograph of a pathology report for a patient. As can be seen in FIG. 1, the pathology of this patient was diagnosed with malignant schwannoma.

FIG. 2 is a photograph of a tumor tissue mass in an immunodeficient mouse with PDX modeling. Wherein the immunodeficient mouse is a full body white hair NOD/Scid mouse. Specifically, subcutaneous tumor formation in the axilla was successful, and primary tumor formation in the axillary nerve sheath was successful.

FIG. 3 is a photograph of the success of primary cell culture on fresh patient tumor tissue blocks. The primary cells can be used for in vitro experiments of malignant schwannoma, but the invention does not use the cells for animal modeling, but uses tissue blocks for in situ modeling and subcutaneous modeling in an immunodeficiency mouse.

The in-situ modeling in PDX has profound significance, the homogeneity of the tumor obtained by in-situ modeling and the primary tumor of a patient is better, the pathogenesis of malignant schwannoma can be deeply discussed after successful modeling, the research and development result can be better repeated in the body of the patient based on the in-situ modeling, meanwhile, a large number of limited malignant schwannoma samples in the body of a clinical patient can be amplified, and a xenograft mouse is used for replacing the clinical patient to carry out different combined drug screening, so that the most appropriate drug is selected, the effectiveness is greatly improved, and the side effect of the drug is reduced. Therefore, the in-situ modeling and application can hopefully improve the effectiveness of clinical malignant nerve sheath tumor treatment and the prognosis of patients, prevent the patients from being disabled and even dying to bring great medical and social burden, reduce the treatment cost and the government financial expenditure, have great practical significance in the aspects and have great economic benefit.

The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions and substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.

Claims

1. A method for constructing a malignant schwannoma xenograft mouse model is characterized by comprising the steps of recovering a malignant schwannoma patient tumor tissue block stored in a liquid nitrogen environment, transplanting the recovered malignant schwannoma patient tumor tissue block into an immunodeficiency mouse under an aseptic condition in a heterogenic subcutaneous mode, selecting an immunodeficiency mouse with the age of 2-6 weeks, enabling the sex of the mouse to be consistent with the sex of the patient, soaking the recovered malignant schwannoma tissue block of the patient in sterile incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, implanting the tumor tissue block of the patient into the subcutaneous part of the mouse, suturing an incision, and feeding the mouse to enable the tumor in the mouse to grow.

2. A method for constructing a malignant schwannoma xenograft mouse model is characterized by comprising the steps of recovering a malignant schwannoma patient tumor tissue block stored in a liquid nitrogen environment, transplanting the recovered malignant schwannoma patient tumor tissue block into an immunodeficiency mouse in a heterogenous and in-situ manner under an aseptic condition, selecting an immunodeficiency mouse with the age of 2-6 weeks, enabling the sex of the mouse to be consistent with the sex of the patient, soaking the recovered malignant schwannoma tissue block of the patient in a sterile incubation liquid containing fetal calf serum, fixing the mouse after anesthesia, transplanting the tumor tissue block of the patient into a nerve sheath of the mouse, suturing an incision, and feeding the mouse to enable the tumor in vivo to grow.

3. The method of claim 1 or 2, further comprising a procedure of programmed cooling and liquid nitrogen preservation and a procedure of rapid warming resuscitation of the collected tumor tissue mass of the malignant schwannoma patient.

4. The method according to claim 3, wherein the tumor tissue mass of the malignant schwannoma patient is preserved in DMEM containing DMSO, FBS and NEAA, the temperature of the three or more stages is reduced to the temperature of liquid nitrogen, and the tumor tissue mass is preserved in the temperature of liquid nitrogen, and the rapid heating resuscitation refers to resuscitation of the frozen tissue in a constant-temperature water bath at 36-38 ℃.

5. The method of claim 1 or 2, wherein said subcutaneous implantation comprises making a skin incision in the mouse axilla, implanting the tumor tissue subcutaneously and suturing the surgical incision; the orthotopic transplantation comprises making a skin incision at the axillary nerve sheath or sciatic nerve sheath of the mouse, separating the nerve sheath and opening, implanting a tumor tissue block of the patient into the nerve sheath, and then suturing the surgical incision.

6. The method according to claim 1 or 2, wherein the weight of the mouse in the subcutaneous transplantation or the orthotopic transplantation is 8-20 g, preferably 12-15g, the sterile incubation solution further comprises an antibiotic, preferably the sterile incubation solution comprises an antibiotic and a DMEM medium or an RPMI-1640 medium, and more preferably the antibiotic comprises penicillin and streptomycin.

7. The method according to claim 1 or 2, wherein the immunodeficient mouse is a NOD/SCID mouse and the unilateral length of the tumor tissue mass of the malignant schwannoma patient implanted in the mouse is less than or equal to 3mm, preferably less than or equal to 1 mm.

8. The method of claim 1 or 2, further comprising observing the size of the malignant schwannoma in the mouse using a small animal in vivo imager, preferably also using a small animal in vivo imager to observe whether the tumor has metastasized to the internal organs of the mouse.

9. The method of claim 1 or 2, further comprising raising the mouse such that the mouse is dissected after growth of the tumor in vivo, detaching the tumor to obtain P0 tumor generation, xenotransplanting or xenotransplanting the P0 tumor generation P1 tumor generation P1 tumor generation P2 tumor generation P in immunodeficient mouse, and so on; preferably the method further comprises the step of programming the P0 generation tumor mass to be cooled and stored in liquid nitrogen, and optionally the steps of programming the P1 and P2 generation tumor mass to be cooled and stored in liquid nitrogen.

10. A method for constructing a mouse model of malignant schwannoma xenograft is characterized in that the method comprises the steps of carrying out xenogeneic subcutaneous transplantation or xenogeneic in-situ transplantation on P0 generation malignant schwannoma tumor tissues, P1 generation malignant schwannoma tumor tissues or P2 generation malignant schwannoma tumor tissues in a mouse model which is frozen in a liquid nitrogen environment and revived under an aseptic condition, soaking malignant schwannoma tumor tissue blocks in sterile incubation liquid containing fetal calf serum, anaesthetizing the mice and then fixing, implanting the malignant schwannoma tumor tissue blocks into the subcutaneous tissues or the nerve sheath of the mice, suturing incisions, and feeding the mice to enable the tumors to grow in vivo.