CN110876359A - Novel mouse in-situ pancreatic cancer model and establishment method thereof - Google Patents
Novel mouse in-situ pancreatic cancer model and establishment method thereof Download PDFInfo
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- 206010061902 Pancreatic neoplasm Diseases 0.000 title claims abstract description 65
- 201000002528 pancreatic cancer Diseases 0.000 title claims abstract description 65
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 title claims abstract description 63
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- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000011065 in-situ storage Methods 0.000 title claims abstract description 14
- 210000000277 pancreatic duct Anatomy 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 14
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- 238000010172 mouse model Methods 0.000 claims abstract description 3
- 210000000496 pancreas Anatomy 0.000 claims description 29
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 28
- 239000010931 gold Substances 0.000 claims description 28
- 229910052737 gold Inorganic materials 0.000 claims description 28
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 21
- 210000001015 abdomen Anatomy 0.000 claims description 20
- 210000000013 bile duct Anatomy 0.000 claims description 18
- 108010082117 matrigel Proteins 0.000 claims description 9
- 238000011580 nude mouse model Methods 0.000 claims description 8
- 241000699660 Mus musculus Species 0.000 claims description 7
- 241000699670 Mus sp. Species 0.000 claims description 7
- 210000003445 biliary tract Anatomy 0.000 claims description 6
- 210000001953 common bile duct Anatomy 0.000 claims description 6
- 210000000813 small intestine Anatomy 0.000 claims description 6
- 206010061598 Immunodeficiency Diseases 0.000 claims description 5
- 208000029462 Immunodeficiency disease Diseases 0.000 claims description 5
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- 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
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- 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
- A01K2207/00—Modified animals
- A01K2207/12—Animals modified by administration of exogenous cells
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- 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
- A01K2227/105—Murine
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- 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
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Abstract
The invention belongs to the technical field of medical treatment, and particularly relates to a novel mouse in-situ pancreatic cancer model and an establishing method thereof. A method for establishing a novel mouse in-situ pancreatic cancer model comprises the step of injecting 100 microliters of a solution containing 200 units of pancreatic cancer cells with 500 ten thousand units into a pancreatic tube end close to a pancreatic nipple, wherein the solution consists of a mature stable pancreatic cancer cell strain and gel liquid. The invention adopts a method of injecting pancreatic cancer cells reversely in a pancreatic duct of a mouse (injecting the pancreatic cancer cells along the pancreatic duct from the head of the mouse to the tail of the mouse) to construct an in-situ pancreatic cancer model of the mouse. The method provided by the invention is simple and effective, and the mouse model established by the invention can provide conditions for the scientific research of clinicians and scientific research workers based on animal models.
Description
Technical Field
The invention belongs to the technical field of medical treatment, and particularly relates to a novel mouse in-situ pancreatic cancer model and an establishing method thereof.
Background
Pancreatic cancer is a progressive, fatal tumor with 216,000 new patients each year; the prognosis is poor, with a survival rate of about 23% 1 year after diagnosis and a 5-year survival rate of no more than 5%. In the united states, there are approximately 46,000 new cases of pancreatic cancer patients, and 39,590 die of pancreatic cancer. Therefore, the research on the pathogenesis of pancreatic cancer and the search for novel therapeutic targets of pancreatic cancer have great significance for the diagnosis and treatment of pancreatic cancer. However, the lack of in situ pancreatic cancer models available at home currently severely restricts the further research on the pathogenesis and new diagnosis and treatment strategies of pancreatic cancer.
Disclosure of Invention
The invention aims to provide a novel method for establishing a mouse in-situ pancreatic cancer model. The method can stably establish an in-situ pancreatic cancer model of a mouse, and is used for researching a pancreatic cancer pathogenesis, researching and developing a novel pancreatic cancer treatment strategy and judging the prognosis of a novel pancreatic cancer treatment method.
In order to achieve the purpose, the novel invention adopts the specific technical scheme that:
a method for establishing a novel mouse in-situ pancreatic cancer model comprises the step of injecting 100 microliters of a solution containing 200 units of pancreatic cancer cells with 500 ten thousand units into a pancreatic tube end close to a pancreatic nipple, wherein the solution consists of a mature stable pancreatic cancer cell strain and gel liquid.
Preferably, the first and second liquid crystal materials are,
the volume ratio of the mature and stable pancreatic cancer cell strain to the matrigel is 1: 4.
The gel liquid was matrigel (Corning, usa).
The method comprises the following specific steps: performing general anesthesia and fixation on balb/c-nu nude mice, clamping a common bile duct close to a pancreas by using a gold clip under a 16-time microscope, and closing the bile duct; finding a pancreatic nipple on the wall of the small intestine, clamping a pancreatic duct of the pancreatic nipple close to the pancreatic end by using a gold clamp, and sealing the pancreatic duct; 100 microliters of pancreatic cancer cells containing 200-; after 20 minutes, opening the gold clip at the nipple of the pancreas to ensure that the pancreatic duct is unblocked, and opening the gold clip at the end of the general bile duct to ensure that the biliary tract is unblocked; and (4) sewing the abdomen of the immunodeficiency mouse by using a 3-0 belly line to complete the model establishment.
Specifically, the existing mature and stable pancreatic cancer cell strain and 20% matrigel (preventing cell backflow) are prepared, balb/c-nu nude mice are anesthetized and fixed systemically, the abdomen is disinfected by alcohol, the abdomen is broken along the midline of the abdomen, and the pancreas and the bile duct are fully exposed. Under a 16-fold microscope, the common bile duct near the pancreas was clamped with a gold clip to close the bile duct. Finding out pancreas papilla on the small intestine wall, clamping the pancreatic duct of the pancreas papilla near the pancreas end by a gold clamp, and sealing the pancreatic duct. 100 microliters of pancreatic cancer cells containing 200-. The immunodeficient mice injected with pancreatic cancer cells were allowed to stand for 20 minutes to allow the cells to settle in the pancreas. After 20 minutes, the gold clip at the nipple of the pancreas is opened first, so that the position where the pancreatic duct is clipped is unblocked, and then the gold clip at the end of the general bile duct is opened, so that the biliary tract is unblocked. And after the pancreatic duct and the bile duct are unobstructed, observing whether pancreatic juice overflows from the pinhole or not. If the mouse does not overflow, the abdomen of the immunodeficiency mouse is sutured by using a 3-0 belly line, and the model building is completed.
The immunodeficient mice injected with pancreatic cancer cells were allowed to stand for 20 minutes and the gold clips at the papilla of the pancreas were opened.
Preferably, it is carried out under 16-fold microscope.
In addition, the invention also provides a mouse in-situ pancreatic cancer model, which is established by injecting 100 microliters of a solution containing 200- & 500 ten thousand units of pancreatic cancer cells into a pancreatic end close to a pancreatic papilla, wherein the solution consists of a mature stable pancreatic cancer cell line and matrigel (Corning corporation, USA), and the volume ratio of the mature stable pancreatic cancer cell line to the matrigel is as follows: 1:4.
Performing general anesthesia and fixation on balb/c-nu nude mice, clamping a common bile duct close to a pancreas by using a gold clip under a 16-time microscope, and closing the bile duct; finding a pancreatic nipple on the wall of the small intestine, clamping a pancreatic duct of the pancreatic nipple close to the pancreatic end by using a gold clamp, and sealing the pancreatic duct; 100 microliters of pancreatic cancer cells containing 200-; after 20 minutes, opening the gold clip at the nipple of the pancreas to ensure that the pancreatic duct is unblocked, and opening the gold clip at the end of the general bile duct to ensure that the biliary tract is unblocked; and (4) sewing the abdomen of the immunodeficiency mouse by using a 3-0 belly line to complete the model establishment.
Specifically, the existing mature and stable pancreatic cancer cell strain and 20% matrigel (preventing cell backflow) are prepared, balb/c-nu nude mice are anesthetized and fixed systemically, the abdomen is disinfected by alcohol, the abdomen is broken along the midline of the abdomen, and the pancreas and the bile duct are fully exposed. Under a 16-fold microscope, the common bile duct near the pancreas was clamped with a gold clip to close the bile duct. Finding out pancreas papilla on the small intestine wall, clamping the pancreatic duct of the pancreas papilla near the pancreas end by a gold clamp, and sealing the pancreatic duct. 100 microliters of pancreatic cancer cells containing 200-. The immunodeficient mice injected with pancreatic cancer cells were allowed to stand for 20 minutes to allow the cells to settle in the pancreas. After 20 minutes, the gold clip at the nipple of the pancreas is opened first, so that the position where the pancreatic duct is clipped is unblocked, and then the gold clip at the end of the general bile duct is opened, so that the biliary tract is unblocked. And after the pancreatic duct and the bile duct are unobstructed, observing whether pancreatic juice overflows from the pinhole or not. If the mouse does not overflow, the abdomen of the immunodeficiency mouse is sutured by using a 3-0 belly line, and the model building is completed.
The immunodeficient mice injected with pancreatic cancer cells were allowed to stand for 20 minutes and the gold clips at the papilla of the pancreas were opened.
Preferably, it is carried out under 16-fold microscope.
In addition, the invention also provides a solution for preparing the mouse in-situ pancreatic cancer model, which consists of a mature and stable pancreatic cancer cell strain and gel liquid; the volume ratio of the pancreatic cancer cell strain to the gel liquid is as follows: 1:4. Preferably, the gel liquid is matrigel.
The invention adopts a method of injecting pancreatic cancer cells reversely in a pancreatic duct of a mouse (injecting the pancreatic cancer cells along the pancreatic duct from the head of the mouse to the tail of the mouse) to construct an in-situ pancreatic cancer model of the mouse. The method provided by the invention is simple and effective, and the mouse model established by the invention can provide conditions for the scientific research of clinicians and scientific research workers based on animal models. The solution 20% gel liquid prevents cell backflow, and facilitates model building.
Drawings
FIG. 1 is a diagram of materials and a specific process for modeling according to the present invention.
FIG. 2 is a graph of the histopathological structure of pancreatic tumors and model anatomical symptoms of the natural progression of pancreatic cancer in this model.
Detailed Description
(1) Preparation of experimental materials: an ultra-clean workbench, a shadowless lamp, a 16-fold microscope (figure 1A), 4% chloral hydrate, a nude mouse fixing plate, a gauze piece, a cotton swab, sterilized alcohol, an insulin needle, a common 1ml syringe, 1 pair of ophthalmic scissors, 2 pairs of micro-ophthalmic curved forceps, 1 pair of micro-needle holding forceps, 1 pair of needle holding devices, 3-0 abdomen closing threads, medical adhesive tape paper, an ophthalmic dilator, 2 gold clips (figure 1B) and pancreatic cancer cells (replaced by Meilan in figure 1C). Balb/c-nu nude mice (FIG. 1D).
(2) The technical scheme adopted by the model is as follows: preparing the existing mature and stable pancreatic cancer cell strain and 20% gel liquid (preventing cell backflow), performing general anesthesia and fixation on balb/c-nu nude mice, disinfecting the abdomen with alcohol, breaking the abdomen along the midline of the abdomen, and fully exposing pancreas and bile duct (fig. 1E). The common bile duct near the pancreas was clamped with gold clips under a 16-fold microscope to close the bile duct (fig. 1F). The papilla of the pancreas was found on the wall of the small intestine, and the pancreatic duct near the pancreatic end of the papilla of the pancreas was clamped with a gold clip to close the pancreatic duct (fig. 1G). 100 microliters of pancreatic cancer cells containing 200-. Immunodeficient mice injected with pancreatic cancer cells were allowed to sit for 20 minutes to pellet the cells in the pancreas (FIG. 1J). After 20 minutes, the gold clip at the nipple of the pancreas is opened first, so that the position where the pancreatic duct is clipped is unblocked, and then the gold clip at the end of the general bile duct is opened, so that the biliary tract is unblocked. After the pancreatic duct and bile duct were unobstructed, the pinhole was observed for rupture and pancreatic juice overflow (fig. 1K). If there was no extravasation, the immunodeficient mice were abdominal sutured using a 3-0 trans-abdominal line to complete the modeling (FIG. 1L).
(3) The natural development process of pancreatic cancer in this model: after the model is established, the model mouse grows to die from the tumor, and the time is about 1 month. Later emaciation, a downward movement of mental status, accompanied by jaundice, is common. After model spontaneous death, the anatomy and symptoms are shown in FIGS. 2A-2C, and the histopathology of the pancreatic tumor is shown in FIG. 2D.
Claims (9)
1. A method for establishing a novel mouse in-situ pancreatic cancer model is characterized by comprising the following steps: 100 microliters of a solution containing 200 units of 500 ten thousand units of pancreatic cancer cells, which consists of a mature stable pancreatic cancer cell line and a gel liquid, was injected into the pancreatic end near the pancreatic papilla.
2. The method of establishing according to claim 1, wherein: the volume ratio of the pancreatic cancer cell strain to the gel liquid is as follows: 1:4.
3. The method of establishing according to claim 1 or 2, characterized in that: the gel liquid is matrigel.
4. The method of establishing according to claim 1 or 2, characterized in that: performing general anesthesia and fixation on balb/c-nu nude mice, clamping a common bile duct close to a pancreas by using a gold clip under a 16-time microscope, and closing the bile duct; finding a pancreatic nipple on the wall of the small intestine, clamping a pancreatic duct of the pancreatic nipple close to the pancreatic end by using a gold clamp, and sealing the pancreatic duct; 100 microliters of pancreatic cancer cells containing 200-; after 20 minutes, opening the gold clip at the nipple of the pancreas to ensure that the pancreatic duct is unblocked, and opening the gold clip at the end of the general bile duct to ensure that the biliary tract is unblocked; and (4) sewing the abdomen of the immunodeficiency mouse by using a 3-0 belly line to complete the model establishment.
5. The method of establishing according to claim 1, wherein: the immunodeficient mice injected with pancreatic cancer cells were allowed to stand for 20 minutes and the gold clips at the papilla of the pancreas were opened.
6. The method of establishing according to claim 1, wherein: performed under 16-fold microscope.
7. A mouse model of orthotopic pancreatic cancer prepared by the method of any one of claims 1 to 6.
8. A solution for making a mouse orthotopic pancreatic cancer model, comprising: consists of a mature and stable pancreatic cancer cell strain and gel liquid; the volume ratio of the pancreatic cancer cell strain to the gel liquid is as follows: 1:4.
9. The method of establishing according to claim 8, wherein: the gel liquid is matrigel.
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| CN201910523378.2A CN110476876A (en) | 2019-06-17 | 2019-06-17 | A kind of novel mouse original position pancreatic cancer models and its method for building up |
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| CN114807014A (en) * | 2022-05-11 | 2022-07-29 | 浙江大学 | Method for relieving pancreatic cancer immunosuppressive microenvironment |
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