CN109392843A - Oophoroma Transplanted tumor model and application are constructed based on organoid method - Google Patents
Oophoroma Transplanted tumor model and application are constructed based on organoid method Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/027—New breeds of vertebrates
- A01K67/0271—Chimeric animals, e.g. comprising exogenous cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0331—Animal model for proliferative diseases
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2531/00—Microcarriers
Abstract
Oophoroma Transplanted tumor model and application are constructed based on organoid method, it belongs to field of medicaments, the purpose of the present invention is with " organoid " culture technique, using microcarrier as timbering material, turn out the ovarian cancer cell -3D composite body in patient source, then it is subcutaneous cell -3D composite body to be directly inoculated in normal immunological mouse, construct the oophoroma Transplanted tumor model in patient source, to solve, current human ovarian cancer Transplanted tumor model is existing to use the immunodeficient mouses such as SCID mouse, nude mice, it is expensive, it is not easy to raise, the problems such as tumor formation rate is not high；Especially solve the problems, such as that immunodeficient mouse human ovarian cancer Transplanted tumor model can not reflect body immune system important function played in tumorigenesis process, and there are the tumor formation period is long for immunodeficient mouse people Transplanted tumor model, it is difficult to solve the clinical susceptibility demand for urgently needing drug therapy patient.The present invention is applied to mouse model and constructs field.
Description
Technical field
The invention belongs to field of medicaments, are based on " organoid " building more particularly to a kind of medicine animal model for tumour-
The normal immunological mouse ovarian cancer Transplanted tumor model in patient source and establish the model method and they in field of medicaments and
Application in clinic.
Background technique
Tumour is a kind of frequently-occurring disease and common disease for seriously threatening human health, and pathogenesis not yet illustrates completely.Ovum
Nest cancer is one of most common malignant tumour, still occupies female according to American Cancer Society's statistical result showed ovarian cancer mortality in 2018
The first place of sexual reproduction system tumor.Therefore by experimental animal model further explore ovarian neoplasm occurrence and development mechanism and
More effective treatment method is significant.
Newly most potential " the year of life science in 2017 is cited as into research achievement organoid (organoids)
Technology ", referred to as organoid technology, organoid are a kind of miniature Three-dimensional cell culture models, are originated from patient's primary tumor, and
It is cultivated in the lab.From organoid formed at the beginning of, to carry out drug therapy after, these organoids histology, point
Son is horizontal and can functionally continue to keep height consistent with source tumor tissues, is well reflected its feature.
For a long time, human tumour cell line and transplantable tumor Nude mice model are used as always tumor research and anti-cancer drugs
Research/development platform, however, lack interstitial cell since cell line is formed by tumour, and generation of the interstitial cell in tumour, hair
It is played an important role in exhibition and transfer process, the research and development of basic research and effective antitumor new drug to tumour cause to be stranded greatly very much
Difficulty, also, transplantable tumor Nude mice model is difficult the true state of an illness of accurate simulation clinical patients, simultaneously, it is difficult to meet preclinical
The needs of experiment.
In recent years, the fresh specimens from pri of patient is transplanted to immunodeficient mouse by foreign scholar, establishes new mould
Type is named as the transplantable tumor (patient-derived xenografts, PDXs) in tumor patient source.Although these models at
Molecular biology, histology and the Pathologic Characteristics of patient tumors are reproduced to function, still, because nude mice lacks T cell, SCID mouse
Lack T cell and B cell simultaneously, it is difficult to illustrate the relationship of tumorigenesis and transfer and immunity of organism, and be not suitable for grinding
Immunotherapy of tumors is studied carefully, nor the most ideal model of exploitation new type antineoplastic medicine.And the normal immunological function in patient source
Mouse transplanting tumor model can overcome the disadvantages that and be unable to accurate simulation body immune system to swollen because immunodeficient mouse lacks immunocyte
The disadvantages of tumor immune attack, so that the more accurately molecular biology characteristics and tumor microenvironment of reduction specimens, are
The pathogenesis and new drug development of research tumour and the clinical ideal model precisely treated.
Summary of the invention
The purpose of the present invention is use microcarrier (NMC2) for timbering material, compound patient with " organoid " culture technique
The ovarian cancer cell in source turns out cell -3D composite body, is then directly inoculated in cell -3D composite body normally
Immune mouse is subcutaneous, the oophoroma Transplanted tumor model in patient source in construct, to solve current human ovarian cancer transplantable tumor mould
Type is existing to be not easy to raise using the immunodeficient mouses such as nude mice, SCID mouse, expensive, the not high problem of tumor formation rate；Especially
It is to solve immunodeficient mouse human ovarian cancer Transplanted tumor model body immune system can not be reflected in tumorigenesis process
Played in important function the problem of and immunodeficient mouse people Transplanted tumor model there are the tumor formation period length (generally require 3-5
A month), it is difficult to solve the clinical susceptibility demand for urgently needing drug therapy patient.
Of the invention constructs oophoroma Transplanted tumor model based on organoid method, it is followed the steps below:
One, it obtains the primary ovarian cancer tumor cell of people: taking ovarian cancer cell and ovarian cancer tissue's interstitial cell, to contain 10%
DMEM, the mass percentage of fetal calf serum are 1% penicillin and mass percentage is the complete culture solution general of 1% streptomysin
Above-mentioned cell carries out trypan blue counting after mixing；If trypan blue living cell counting number > 95%, adjust cell concentration to 2 ×
107/ mL is put into 37 DEG C, 5%CO after being inoculated into culture plate or culture bottle2Incubator obtains the primary ovarian cancer cell suspension of people；
Two, by microcarrier be soaked in volumn concentration be 75% alcohol for 24 hours after, cleaned 4 times with 1 × PBS buffer solution,
It is placed in 24~72h of incubation in DMEM culture medium；People primary ovarian cancer cell suspension and micro- load for being incubated for by step 1 obtains
Body mixing is 37 DEG C, 5%CO in temperature212~48h, cell-microcarrier complex after being incubated for are cultivated in incubator；
Three, cell-microcarrier complex 4 times after being incubated for 1 × PBS buffer solution cleaning step two, then it is slow with 1 × PBS
Fliud flushing is resuspended cell and adjusts cell concentration to 2 × 107/ mL adjusts microcarrier concentration to 250~300ug/mL, it is micro- to obtain cell-
Vehicle suspension is placed in spare on ice；Cell-microcarrier suspension is injected in 7~8 weeks normal immunological Mice Bodies, body
After being implanted into 20~30 days, that is, construct the oophoroma transplantable tumor mouse model for completing patient source.
The present invention include it is following the utility model has the advantages that
Invention introduces tumour 3 D culture techniques, utilize patient source in " organoid " dimensional culture technology construct
Ovarian Cancer Model, primary ovarian cancer cell is mainly inoculated in co-incubation on microcarrier (NMC2) timbering material, then
Directly tumor model will be constructed in cell-composite body injection normal immunological Mice Body.
The present invention utilizes " organoid " culture technique, with microcarrier (NMC2) for base material, the original in compound patient source
Immune function normal mouse, which is inoculated in, for ovarian cancer cell subcutaneously successfully constructs the mouse patient with normal immunological function
The oophoroma neoplasm transplantation in source.Compared with existing immunodeficient mouse neoplasm transplantation, which can be better
Study and further elucidate tumour occur in immune function normal body, development mechanism, while also being mentioned for cancer drug development
Relatively previous more valuable new animal model is supplied.
Ovarian cancer cell-the microcarrier complex in patient source is seeded to immune function normal mouse by the present invention, is connect certainly
Starting after kind to the 8-10 days, mouse ingests normally, and activity situation is good, generally starts at the 10th day, and mouse activity is slightly reduced, but
Appetite, hair, weight no significant difference.8~10d, that is, accessible tumour after inoculation, tumor formation rate are 80% (16/20),
Transplantable tumor form is relatively regular, mostly round or ellipse, and color is based on canescence or bois de rose.
Detailed description of the invention
Fig. 1 is the primary ovarian cancer cell dimensional culture system figure of people being successfully established in vitro；Wherein, A: empty carrier (100
×) figure；B: primary ovarian cancer cell culture (200 ×) figure for 24 hours；C: primary ovarian cancer cell is schemed for 24 hours with NMC2 co-incubation；
Fig. 2 is mice with tumor gross examination of skeletal muscle figure；
Fig. 3 is transplantable tumor gross examination of skeletal muscle figure；
Fig. 4 is the oophoroma transplantable tumor HE colored graph in patient source；Wherein, A and B indicates that Primary Tumor, C and D indicate to move
Plant tumor；
Fig. 5 is the oophoroma transplantable tumor immunohistochemical staining figure in patient source.
Specific embodiment
Specific embodiment 1: present embodiment based on organoid method construct oophoroma Transplanted tumor model, it be by
It is carried out according to following steps:
One, it obtains the primary ovarian cancer tumor cell of people: taking ovarian cancer cell and ovarian cancer tissue's interstitial cell, to contain 10%
DMEM, the mass percentage of fetal calf serum are 1% penicillin and mass percentage is the complete culture solution general of 1% streptomysin
Above-mentioned cell carries out trypan blue counting after mixing；If trypan blue living cell counting number > 95%, adjust cell concentration to 2 ×
107/ mL is put into 37 DEG C, 5%CO after being inoculated into culture plate or culture bottle2Incubator obtains the primary ovarian cancer cell suspension of people；
Two, by microcarrier be soaked in volumn concentration be 75% alcohol for 24 hours after, cleaned 4 times with 1 × PBS buffer solution,
It is placed in 24~72h of incubation in DMEM culture medium；People primary ovarian cancer cell suspension and micro- load for being incubated for by step 1 obtains
Body mixing is 37 DEG C, 5%CO in temperature212~48h, cell-microcarrier complex after being incubated for are cultivated in incubator；
Three, cell-microcarrier complex 4 times after being incubated for 1 × PBS buffer solution cleaning step two, then it is slow with 1 × PBS
Fliud flushing is resuspended cell and adjusts cell concentration to 2 × 107/ mL adjusts microcarrier concentration to 250~300ug/mL, it is micro- to obtain cell-
Vehicle suspension is placed in spare on ice；Cell-microcarrier suspension is injected in 7~8 weeks normal immunological Mice Bodies, body
After being implanted into 20~30 days, that is, construct the oophoroma transplantable tumor mouse model for completing patient source.
Specific embodiment 2: the present embodiment is different from the first embodiment in that: the microcarrier is by more
The poroid bar rope sample of layer be cross-linked with each other curl into can positive electrochemical organic composite polymer composition, contain " labyrinth " sample irregular structure
Body.It is other same as the specific embodiment one.
Specific embodiment 3: the present embodiment is different from the first embodiment in that: the people that step 1 obtains is primary
The volume ratio of ovarian cancer cell suspension and the microcarrier being incubated for is 3~4:1.It is other same as the specific embodiment one.
Specific embodiment 4: the present embodiment is different from the first embodiment in that: the mouse is immune function
It can normal mouse.It is other same as the specific embodiment one.
Specific embodiment 5: the present embodiment is different from the first embodiment in that: the mouse is immune function
Energy normal mouse refers to C57BL/6 mouse or BALB/c mouse.It is other same as the specific embodiment one.
Specific embodiment 6: the present embodiment is different from the first embodiment in that: ovarian cancer cell and oophoroma
Organizing interstitial cell is by 20~30g of Qu Xian ovarian cancer tissue, is the collagen of 0.025%-0.05% with mass percentage
Protease digestion separation, obtains ovarian cancer cell and ovarian cancer tissue's interstitial cell.It is other same as the specific embodiment one.
Specific embodiment 7: the present embodiment is different from the first embodiment in that: cell-microcarrier in step 3
The position that suspension injects mouse is subcutaneous under right axillary, every 100~150uL of injection.It is other same as the specific embodiment one.
Specific embodiment 8: the application of the oophoroma Transplanted tumor model using the building of the building of specific embodiment one,
It is for ovarian tumors Mechanism Study and ovarian cancer research and development etc..
Specific embodiment 9: present embodiment is unlike specific embodiment eight: it is in ovarian cancer resistance medicament
Clinical application in screening.It is other identical as specific embodiment eight.
The content of present invention is not limited only to the content of the respective embodiments described above, the group of one of them or several specific embodiments
The purpose of invention also may be implemented in contract sample.
Beneficial effects of the present invention are verified by following instance:
Embodiment 1
Step 1: the acquisition of human ovarian cancer primary cell
The acquisition of the primary ovarian cancer cell in patient source: in patient's informed consent, operation is taken to cut off fresh
Ovarian cancer tissue 20-30g separates ovarian cancer cell with mass percentage for the collagenase digestion of 0.025%-0.05%
With ovarian cancer tissue's interstitial cell, with the DMEM containing 10% fetal calf serum, mass percentage be 1% penicillin and quality hundred
The cell mixing for dividing content to obtain digestion separation for 1% streptomysin complete culture solution, trypan blue living cell counting number > 95%,
Adjusting cell concentration is 2 × 107/ mL puts 37 DEG C, 5%CO2Incubator is spare.
Step 2: the preparation of liquid phase microcarrier
Microcarrier NMC2 is soaked in 75% alcohol 48h, 1 × phosphate buffer (PBS) cleans 4 times, cultivates in DMEM
It is incubated in base spare for 24 hours.
Step 3: the foundation of three-dimensional nodule cell culture model
By the ovarian cancer cell of trypan blue living cell counting number > 95% and ovarian cancer tissue's interstitial cell suspension and it is incubated for
Microcarrier (NMC2) mix (volume ratio 3-4:1), be made containing 2 × 107The cell of a cell/mL-microcarrier suspension；37
DEG C, 5%CO2It is cultivated 24-48 hours in incubator, cell is made to reach saturation shape in microcarrier (NMC2).
Step 4: the building of the oophoroma normal immunological mice-transplanted tumor in patient source
Cell-microcarrier complex 3 times be incubated for are cleaned with 1 × PBS liquid, then the micro- load of cell-is resuspended with 1 × PBS liquid
(cell concentration is 2 × 10 to composite7/ mL, microcarrier concentration 250ug/mL), by the made cell-microcarrier got ready
(NMC2) suspension is placed on spare on ice；It is normal that cell-microcarrier complex is then injected in 7-8 weeks big immune function
Mouse, position are subcutaneous under right axillary, every 100uL-150uL, and the subcutaneous hillock that formed indicates to be inoculated with successfully, et al. Ke 20-
30 days, construct the oophoroma Transplanted tumor model in normal immunological mouse patient source.
Step 5: mouse tumor formation situation
Observe the ordinary circumstances such as spirit, diet, activity, the stool and urine of mouse after transplanting daily, at transplanting subcutaneously whether at
Tumor.From from inoculation to inoculation position occur it is tangible to solid mass when for incubation period, find to start after tumor formation daily
With the maximum gauge (Dmax) and minimum diameter (Dmin) of vernier caliper measurement tumour, by formula V=1/2 × Dmax × (Dmin)
2 calculate gross tumor volume (as depicted in figs. 1 and 2).
Step 6: pathological examination
Tumor growth 20-30 days, cervical dislocation put to death mouse, took out complete tumors, and 10% neutral formalin is fixed, and made
Make paraffin section, Hematoxylin-eosin (HE) dyeing, immunohistochemistry detection P63, P40, CK56 be mainly expressed in cancer cell membrane and
Cytoplasm.
Mouse ordinary circumstance and growth of transplanted human situation
To the 10th day since after inoculation, mouse ingests normally, and activity situation is good, and the 10th day starts, and mouse activity slightly subtracts
It is few, but appetite, hair, weight no significant difference.The can touch tumour after inoculation 5-7 days, and tumour is raw within the 8-15 days
It is long quick, the decline of late growth speed.Transplantable tumor is mostly ellipse, and surface imperfection is more visible with boundary around, is in lark
(see Fig. 3).
Three-dimensional originally culture system
Microscopic observation NMC2 is under the microscope spindle shape shape, and quality is loose, is similar to fibr tissue (such as Figure 1A)；2D group ovary
It can see cancer cell after cancer cell adherent growth (such as Figure 1B), primary ovarian cancer cell and NMC2 co-cultivation 2h preferably to attach
On NMC2, saturation state (such as Fig. 1 C) is basically reached.
Oncological pathology
It is obvious, disordered arrangements swollen that HE dyes the big deep dye of transplantable tumor tissue paraffin section de Visible Core under light microscopic, heteromorphism
Oncocyte；Foreign body reaction (such as Fig. 4) in interstitial around visible big amount lymphocyte and the NMC2 not yet removed.
Transplantable tumor immunohistochemical staining
Immunohistochemical staining P63, P40, CK56 are mainly expressed in cancer cell membrane and cytoplasm, as the result is shown P63, P40,
CK56 is positive expression (such as Fig. 5), and P63, P40, CK56 are positive expression as the result is shown, further confirms tumour cell
In the presence of.
By above-mentioned experiment it is found that the model that the present invention establishes is the oophoroma normal immunological function murine heterotropic in patient source
Neoplasm transplantation has method easy, and the period is short, is easy to raise, the high advantage of tumor formation rate.The model can be studied preferably
With further elucidate oophoroma in immune function normal body and occur, development mechanism, ovarian cancer resistance susceptibility detection, while being also
The research and development of ovarian cancer resistance medicament provide relatively previous more valuable new animal model.
Claims (9)
1. constructing oophoroma Transplanted tumor model based on organoid method, it is characterised in that it is followed the steps below:
One, it obtains the primary ovarian cancer tumor cell of people: taking ovarian cancer cell and ovarian cancer tissue's interstitial cell, to contain 10% tire ox
DMEM, the mass percentage of serum are 1% penicillin and mass percentage is that the complete culture solution of 1% streptomysin will be above-mentioned
Cell carries out trypan blue counting after mixing；If trypan blue living cell counting number > 95%, adjusts cell concentration to 2 × 107/ mL,
37 DEG C, 5%CO are put into after being inoculated into culture plate or culture bottle2Incubator obtains the primary ovarian cancer cell suspension of people；
Two, by microcarrier be soaked in volumn concentration be 75% alcohol for 24 hours after, cleaned 4 times, be placed in 1 × PBS buffer solution
24~72h is incubated in DMEM culture medium；The primary ovarian cancer cell suspension of people that step 1 obtains is mixed with the microcarrier being incubated for
It closes, is 37 DEG C, 5%CO in temperature212~48h, cell-microcarrier complex after being incubated for are cultivated in incubator；
Three, cell-microcarrier complex 4 times after being incubated for 1 × PBS buffer solution cleaning step two, then with 1 × PBS buffer solution
Cell is resuspended and adjusts cell concentration to 2 × 107/ mL adjusts microcarrier concentration to 250~300ug/mL, obtains cell-microcarrier
Suspension is placed in spare on ice；Cell-microcarrier suspension is injected in 7~8 weeks normal immunological Mice Bodies, is planted in vivo
After entering 20~30 days, that is, construct the oophoroma transplantable tumor mouse model for completing patient source.
2. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that described
Microcarrier be cross-linked with each other by the poroid bar rope sample of multilayer curl into can the organic composite polymer of positive electrification form, containing " fan
Palace " sample irregular structure body.
3. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that step 1
The volume ratio of the primary ovarian cancer cell suspension of obtained people and the microcarrier being incubated for is 3~4:1.
4. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that described
Mouse is the normal mouse of immune function.
5. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that described
Mouse is C57BL/6 mouse or BALB/c mouse.
6. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that oophoroma
Cell and ovarian cancer tissue's interstitial cell are by 20~30g of Qu Xian ovarian cancer tissue, are 0.025%- with mass percentage
0.05% collagenase digestion separation, obtains ovarian cancer cell and ovarian cancer tissue's interstitial cell.
7. according to claim 1 construct oophoroma Transplanted tumor model based on organoid method, it is characterised in that step 3
Middle cell-microcarrier suspension injection mouse position is subcutaneous under right axillary, every 100~150uL of injection.
8. such as the application for the oophoroma Transplanted tumor model that claim 1 constructs, it is characterised in that it is ground for ovarian cancer
Hair.
9. application according to claim 8, it is characterised in that it is the clinical application in ovarian cancer resistance medicament screening.
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| CN111607568A (en) * | 2020-05-06 | 2020-09-01 | 苏州济研生物医药科技有限公司 | Culture method of primary ovarian cancer cells and application of primary ovarian cancer cells in drug screening |
| CN113151175A (en) * | 2020-04-12 | 2021-07-23 | 江苏安泰康健康科技有限公司 | Method for shortening period of human tumor xenograft PDX model |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110484506A (en) * | 2019-08-27 | 2019-11-22 | 中南大学湘雅医院 | The construction method of glioblastoma organoid model and application |
| CN110484506B (en) * | 2019-08-27 | 2021-04-02 | 中南大学湘雅医院 | Construction method and application of glioblastoma organoid model |
| CN111040996A (en) * | 2019-12-06 | 2020-04-21 | 北京科途医学科技有限公司 | Method for preparing ovarian cancer organoid |
| CN113151175A (en) * | 2020-04-12 | 2021-07-23 | 江苏安泰康健康科技有限公司 | Method for shortening period of human tumor xenograft PDX model |
| CN111607568A (en) * | 2020-05-06 | 2020-09-01 | 苏州济研生物医药科技有限公司 | Culture method of primary ovarian cancer cells and application of primary ovarian cancer cells in drug screening |
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