Type 2 diabetes mellitus mouse pancreatic cancer model construction method based on living body imaging technology

A type 2 diabetes, in vivo imaging technology, applied in the biological field, can solve the problems of inability to judge metastases, rough modeling methods, and lack of spatial distribution of tumor formation.

Pending Publication Date: 2021-03-26
广西医科大学附属武鸣医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this traditional modeling method is relatively rough, and its tumor formation can only be judged by the naked eye. It is easy to miss the best observation time points of early tumor formation and early metastasis, and the spatial distribution of tumor formation is also lacking in precise positioning. Judging metastases that are invisible to the naked eye can easily lead to deviations in research results
Moreover, due to the long time and difficulty of modeling type 2 diabetes, the previous tumor-bearing models of diabetes were all modeled on the basis of type 1 diabetes, but type 2 diabetes is closely related to pancreatic cancer. Ideal type 2 diabetes pancreatic cancer animal model

Method used

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Embodiment 1

[0020] A method for constructing a pancreatic cancer model in mice with type 2 diabetes based on in vivo imaging technology, comprising the following steps:

[0021] (1) Cell culture: human pancreatic cancer tumor cells were placed in 37°C, 5% CO2 medium, and 1640 medium containing 10% fetal bovine serum and 1% double antibody contained 100ug / ml penicillin and 100ug / ml streptomycin.

[0022] (2) Cell transfection: Take human-derived pancreatic cancer tumor cells in a good logarithmic growth state, spread them in culture flasks, and ensure that the cells reach 20% confluence. Take 4ml of the culture medium containing luciferase-labeled retrovirus, add 40ul (10mg / ml) polybrene to mix, filter through a 0.45um filter membrane, add to human pancreatic cancer tumor cells, incubate overnight at 37°C and replace with complete medium. 8ug / ml blasticidin (Blasticidin S) was added to select stably infected cell clones. Three days after infection, the fluorescence expression was observ...

Embodiment 2

[0027] A method for constructing a pancreatic cancer model in mice with type 2 diabetes based on in vivo imaging technology, comprising the following steps:

[0028] (1) Cell culture: human pancreatic cancer tumor cells were placed in 37°C, 5% CO2 medium, and 1640 medium containing 10% fetal bovine serum and 1% double antibody contained 100ug / ml penicillin and 100ug / ml streptomycin.

[0029](2) Cell transfection: Take human-derived pancreatic cancer tumor cells in a good logarithmic growth phase, spread them in culture flasks, and ensure that the cells reach 30% confluence. Take 4ml of the culture medium containing luciferase-labeled retrovirus, add 40ul (10mg / ml) polybrene to mix, filter through a 0.45um filter membrane, add to human pancreatic cancer tumor cells, incubate overnight at 37°C and replace with complete medium. 8ug / ml blasticidin (Blasticidin S) was added to select stably infected cell clones. Four days after infection, the fluorescence expression was observed...

Embodiment 3

[0034] A method for constructing a pancreatic cancer model in mice with type 2 diabetes based on in vivo imaging technology, comprising the following steps:

[0035] (1) Cell culture: human pancreatic cancer tumor cells were placed in 37°C, 5% CO2 medium, and 1640 medium containing 10% fetal bovine serum and 1% double antibody contained 100ug / ml penicillin and 100ug / ml streptomycin.

[0036] (2) Cell transfection: Take human-derived pancreatic cancer tumor cells in a good logarithmic growth phase, spread them in culture flasks, and ensure that the cells reach 25% confluence. Take 4ml of the culture medium containing luciferase-labeled retrovirus, add 40ul (10mg / ml) polybrene to mix, filter through a 0.45um filter membrane, add to human pancreatic cancer tumor cells, incubate overnight at 37°C and replace with complete medium. 8ug / ml blasticidin (Blasticidin S) was added to select stably infected cell clones. Three days after infection, the fluorescence expression was observ...

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Abstract

The invention discloses a type 2 diabetes mellitus mouse pancreatic cancer model construction method based on a living body imaging technology. The method comprises the following steps: culturing human-derived pancreatic cancer tumor cells, and then carrying out cell transfection to construct a pancreatic cancer cell strain capable of stably expressing luciferase genes; establishing a type 2 diabetes mellitus mouse model, then establishing a type 2 diabetes mellitus mouse pancreatic cancer model, finally carrying out living body bioluminescence imaging observation, dynamically observing tumorgrowth and metastasis conditions in a model mouse by using an animal living body imaging system, then placing the model mouse in the living body imaging system for exposure imaging, and collecting photon numerical values of the model mouse; and drawing a growth curve of the tumor cells in the model mouse. The method disclosed by the invention provides an ideal experimental animal model for researching a molecular mechanism from blood glucose regulation of pancreatic cancer to early metastasis in vivo, and opens up a new field for preparing animal models for clinically treating chronic diseasescomplicated with malignant tumors.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a type 2 diabetic mouse pancreatic cancer model based on live imaging technology. Background technique [0002] Diabetes is closely related to pancreatic cancer, however, the mechanism by which diabetes leads to tumor cell growth and metastasis in pancreatic cancer remains unclear. For in-depth research, ideal experimental animal models are urgently needed. At present, animal models of pancreatic cancer can be roughly divided into the following three categories: (1) chemically induced animal models of pancreatic cancer, which are characterized by convenient preparation, but the cycle is long, and the induction success rate is not high, and the pathological characteristics of induced tumors are similar to those of humans. Tumors vary widely. (2) Human-derived adenocarcinoma mouse inoculation animal model, one is conventional subcutaneous inoculation to form...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N5/09C12N15/53A01K67/027A61B5/00A61D1/00A61D7/00
CPCA01K67/0275A01K2227/105A01K2267/0337A61B5/0071A61B2503/40A61B2503/42A61D1/00A61D7/00C12N5/0693C12N15/52
Inventor 秦雯黄勇
Owner 广西医科大学附属武鸣医院
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