CN113317272A - Rabbit liver transplantation tumor model of VX2 - Google Patents
Rabbit liver transplantation tumor model of VX2 Download PDFInfo
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- CN113317272A CN113317272A CN202110647700.XA CN202110647700A CN113317272A CN 113317272 A CN113317272 A CN 113317272A CN 202110647700 A CN202110647700 A CN 202110647700A CN 113317272 A CN113317272 A CN 113317272A
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- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 68
- 241000283973 Oryctolagus cuniculus Species 0.000 title claims abstract description 44
- 210000004185 liver Anatomy 0.000 title claims abstract description 22
- 238000002054 transplantation Methods 0.000 title claims abstract description 15
- 230000005740 tumor formation Effects 0.000 claims abstract description 12
- 238000010255 intramuscular injection Methods 0.000 claims abstract description 8
- 239000007927 intramuscular injection Substances 0.000 claims abstract description 8
- 210000003205 muscle Anatomy 0.000 claims abstract description 8
- 210000001519 tissue Anatomy 0.000 claims abstract description 8
- 230000003444 anaesthetic effect Effects 0.000 claims abstract description 6
- 238000010276 construction Methods 0.000 claims abstract description 5
- 210000000689 upper leg Anatomy 0.000 claims abstract description 5
- 206010002091 Anaesthesia Diseases 0.000 claims abstract description 4
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 claims abstract description 4
- 230000037005 anaesthesia Effects 0.000 claims abstract description 4
- 229960002275 pentobarbital sodium Drugs 0.000 claims abstract description 3
- 210000000683 abdominal cavity Anatomy 0.000 abstract description 7
- 238000002513 implantation Methods 0.000 abstract description 3
- 206010027476 Metastases Diseases 0.000 abstract description 2
- 208000015181 infectious disease Diseases 0.000 abstract description 2
- 230000009401 metastasis Effects 0.000 abstract description 2
- 208000014018 liver neoplasm Diseases 0.000 description 13
- 201000007270 liver cancer Diseases 0.000 description 10
- 230000036770 blood supply Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 206010019695 Hepatic neoplasm Diseases 0.000 description 3
- 210000001015 abdomen Anatomy 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003533 narcotic effect Effects 0.000 description 3
- 238000010171 animal model Methods 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 210000002767 hepatic artery Anatomy 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 238000007918 intramuscular administration Methods 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 101000777461 Homo sapiens Disintegrin and metalloproteinase domain-containing protein 17 Proteins 0.000 description 1
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 210000003815 abdominal wall Anatomy 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000010109 chemoembolization Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000002951 depilatory effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 102000048999 human ADAM17 Human genes 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229960004194 lidocaine Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 238000002690 local anesthesia Methods 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000017708 myomatous neoplasm Diseases 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 230000024121 nodulation Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- 210000003240 portal vein Anatomy 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011277 treatment modality Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/107—Rabbit
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a VX2 rabbit liver transplantation tumor model. The construction method of the VX2 rabbit liver transplantation tumor model comprises the following steps: firstly, preparing an anesthetic for intramuscular injection, wherein the concentration of the anesthetic is 3% of pentobarbital sodium, and the dosage is 1 mL/kg; then VX2 tumor mass for intramuscular injection was treated to 1X 1mm3The tumor mass with the size is inoculated in the body of a young rabbit under the anesthesia state to form tumor, and the tumor forming part is the thigh muscle part of the two sides of the young rabbit; finally, the separated VX2 tumor tissue is rapidly stored in PBS at 4 ℃, muscle tissue around the tumor is removed by a cell operating platform, and the tumor block is cut into 1 multiplied by 1mm3After the size of the rabbit is transplanted into the liver of a healthy adult rabbit for planting. The VX2 rabbit liver transplantation tumor model disclosed by the invention is high in tumor formation rate, the tumor formation usually presents a single tumor, and the probability of abdominal cavity implantation metastasis and abdominal cavity infection is low, so that the subsequent experimental study is facilitated.
Description
Technical Field
The invention relates to a VX2 rabbit liver transplantation tumor model, and belongs to the technical field of biological medicines.
Background
Hepatocellular carcinoma (HCC) is one of the most common malignancies, with global incidence ranking fifth in men, second in tumor mortality, seventh in women and sixth in tumor mortality. About 85% of these cases occur in developing countries, especially in the areas (southeast Asia and Africa) where Hepatitis B Virus (HBV) is prevalent, and the number of cases of liver cancer in China accounts for 46.6% of the worldwide cases of liver cancer. The early liver cancer usually has no obvious symptoms, except that a few early liver cancer patients can receive radical treatment, most liver cancer patients are in the middle and late stages when discovered, and the chance of radical operation is lost. Transcatheter Arterial Chemoembolization (TACE) is The most widely used treatment modality for patients with advanced Liver Cancer in The Barcelona Clinical Liver Cancer (BCLC) stage. Unlike normal liver tissue, liver cancer usually has about 90% of its blood supply from the hepatic artery and only less than 10% from the portal vein, which provides a theoretical basis for TACE treatment. In the human TACE simulation surgery, the rabbit VX2 liver tumor model has irreplaceable advantages and is the first choice animal model for the current interventional field experimental study.
The VX2 cell strain is a tumor cell line which is induced by a Shope virus to rabbit skin squamous carcinoma cells and is established by dozens of passages, and is characterized by mainly comprising: the tumor growth rate is high, the tumor growth speed is high, the tumor is rich in blood and low in differentiation, and the pathological type is mostly squamous carcinoma. The rabbit liver transplantable tumor is similar to human liver cancer blood supply, mainly takes hepatic artery blood supply as main blood supply, and is a common tumor-bearing animal model for developing liver cancer interventional therapy experiments. Currently, the commonly used tumor breeding methods for the rabbit VX2 liver tumor model include: tumor mass tumor under direct vision after opening abdomen, and tumor cell suspension injection under the guidance of percutaneous ultrasound and CT. The problems with the current experimental approach, open abdominal neoplasia: the time is long, and the skilled operation is generally performed for 1h from opening the abdomen to suturing 1 rabbit; the abdominal cavity has large lesion of the seed tumor, abdominal wall planting and transfer are easy to occur after operation, the later intervention experiment is greatly influenced, and the specimen drawing of the experiment endpoint is influenced finally. The puncture under the ultrasonic guidance and the injection mode of cell suspension have the problems of low tumor formation rate, poor single tumor formation effect, easy occurrence of abdominal cavity scattering and the like.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the rabbit VX2 liver tumor model in the prior art has the problems of many complications, low tumor formation rate, poor single tumor formation effect, easy occurrence of abdominal cavity dissemination, time-consuming operation of a tumor planting method and the like.
In order to solve the technical problems, the invention provides a construction method of a VX2 rabbit liver transplantation tumor model, which comprises the following steps:
step 1: preparing an anesthetic for intramuscular injection, wherein the concentration of the anesthetic is 3% of pentobarbital sodium, and the dosage is 1 mL/kg;
step 2: VX2 tumor mass for intramuscular injection was treated to 1X 1mm3The tumor mass with the size is inoculated in the body of a young rabbit under the anesthesia state to form tumor, and the tumor forming part is the thigh muscle part of the two sides of the young rabbit;
and step 3: taking out VX2 tumor tissue obtained by the tumor formation in the step 2, rapidly storing the tumor tissue in PBS at 4 ℃, removing muscle tissue around the tumor by a cell operating platform to obtain tumor blocks, and cutting the tumor blocks into 1 × 1 × 1mm3After the size of the rabbit is transplanted into the liver of a healthy adult rabbit for planting.
The invention also provides a VX2 rabbit liver transplantation tumor model which is constructed by adopting the construction method of the VX2 rabbit liver transplantation tumor model.
Compared with the prior art, the invention has the beneficial effects that:
the VX2 rabbit liver transplantation tumor model disclosed by the invention is high in tumor formation rate, the tumor formation usually presents a single tumor, and the probability of abdominal cavity implantation metastasis and abdominal cavity infection is low, so that the subsequent experimental study is facilitated.
Detailed Description
In order to make the invention more comprehensible, preferred embodiments are described in detail below.
A VX2 rabbit liver transplantation tumor model is constructed by the following steps:
step (1): tumor-bearing rabbit tumorigenicity of VX2 tumor mass
First, young rabbits (young new zealand white rabbits of about 1000g size) are selected as tumor-bearing rabbits, and the immune system of the young rabbits is not complete and easy to make muscle tumor. Rabbit narcotic preparation, wherein the concentration of the sodium pentobarbital powder is 3% (3g/100ml NS), the dosage of the narcotic is 1ml/kg, and the rabbit narcotic takes effect after intramuscular injection for about 10 min; the advantages are that: firstly, the operation is simple, and the operation can be quickly finished by only one person without special fixation; secondly, it is more important than intravenous injection that the rabbit does not die because the drug enters the blood too quickly; finally, 3 percent of the preparation is adopted, the total amount of intramuscular injection liquid is small, and the intramuscular edema can not be caused. After the anesthesia takes effect, the rabbit hair on the thighs on both sides is shaved and removed, and the depilatory cream can be used for depilating on condition, so that the influence of the skin hair in the operation area is reduced; after shaving, the local skin is disinfected.
Then, the tumor mass was cut into 1X 1mm3Sucking large and small tumor masses into a tube by using a 1ml syringe, puncturing the skin by using an 18G needle, enabling the puncture needle to enter the thigh muscles of rabbits at two sides, wherein the depth of the puncture needle is 0.5cm, and locally compressing and stopping bleeding after the injection of the large and small tumor masses; waiting for 2 weeks for nodulation.
Step (2): intrahepatic implantation of VX2 tumor mass
After waiting for 2 weeks, the diameter of the intramuscular tumor was about 1cm, after local re-depilation and sterilization, the skin was cut open, the surrounding tissue was isolated, the tumor was dissociated and rapidly stored in PBS at 4 ℃, the muscle tissue around the tumor was removed in a cell platform, and the tumor mass was cut into pieces of 1X 1mm3The size is reserved; healthy rabbits (about 3.5kg adult New Zealand white rabbits) are anesthetized, and after belly unhairing and disinfection, local lidocaine local anesthesia at puncture points; the tumor mass is sent into the head end of an ultrasonic puncture needle through an ophthalmic forceps, the left lobe of a rabbit liver is punctured under the guidance of ultrasonic, a thick tissue area is taken close to the hepatic portal, the puncture needle enters the liver with the depth of 1cm, after the position is confirmed, the tumor mass is sent into an inner core of the ultrasonic puncture needle, the tumor tissue in the needle is pushed into the liver parenchyma, the puncture needle is pulled out, and hemostasis by compression is performed. Waiting for 2 weeks for tumor formation, and obtaining a VX2 rabbit liver transplantation tumor model after tumor formation.
The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way and substantially, it should be noted that those skilled in the art may make several modifications and additions without departing from the scope of the present invention, which should also be construed as a protection scope of the present invention.
Claims (2)
1. A construction method of a VX2 rabbit liver transplantation tumor model is characterized by comprising the following steps:
step 1: preparing an anesthetic for intramuscular injection, wherein the concentration of the anesthetic is 3% of pentobarbital sodium, and the dosage is 1 mL/kg;
step 2: VX2 tumor mass for intramuscular injection was treated to 1X 1mm3The tumor mass with the size is inoculated in the body of a young rabbit under the anesthesia state to form tumor, and the tumor forming part is the thigh muscle part of the two sides of the young rabbit;
and step 3: taking out VX2 tumor tissue obtained by the tumor formation in the step 2, rapidly storing the tumor tissue in PBS at 4 ℃, removing muscle tissue around the tumor by a cell operating platform to obtain tumor blocks, and cutting the tumor blocks into 1 × 1 × 1mm3After the size of the rabbit is transplanted into the liver of a healthy adult rabbit for planting.
2. A VX2 rabbit liver transplantation tumor model, which is constructed by the construction method of the VX2 rabbit liver transplantation tumor model of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110647700.XA CN113317272A (en) | 2021-06-10 | 2021-06-10 | Rabbit liver transplantation tumor model of VX2 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110647700.XA CN113317272A (en) | 2021-06-10 | 2021-06-10 | Rabbit liver transplantation tumor model of VX2 |
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| CN113317272A true CN113317272A (en) | 2021-08-31 |
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| CN202110647700.XA Pending CN113317272A (en) | 2021-06-10 | 2021-06-10 | Rabbit liver transplantation tumor model of VX2 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2799132C1 (en) * | 2023-03-28 | 2023-07-04 | федеральное государственное бюджетное учреждение "Национальный медицинский исследовательский центр онкологии" Министерства здравоохранения Российской Федерации | Method of creating liver cancer model on immunodeficient mice |
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2021
- 2021-06-10 CN CN202110647700.XA patent/CN113317272A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2799132C1 (en) * | 2023-03-28 | 2023-07-04 | федеральное государственное бюджетное учреждение "Национальный медицинский исследовательский центр онкологии" Министерства здравоохранения Российской Федерации | Method of creating liver cancer model on immunodeficient mice |
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Application publication date: 20210831 |
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