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Method for establishing green fluorescence BALB/c nude mouse model and model application

A green fluorescence, mouse model technology, used in biochemical equipment and methods, microbial assay/inspection, animal husbandry, etc. Environmental cells, unstable genetic traits, etc.

Inactive Publication Date: 2017-12-08
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the research on such animal models is relatively mature, there are certain defects: the tumor model established with ordinary BALB / c nude mice cannot track and distinguish the tumor and the tumor microenvironment cells derived from the host; The fluorescent nude mice crossed between nude mice and fluorescent mice with normal immune function, because of their unstable genetic traits, the fluorescent expression of their offspring is not stable
[0004] At present, transgenic green fluorescent mice can emit green fluorescence because of their whole body tissues and organs, so they are suitable for studying the interaction between exogenous tissues or cells (usually another fluorescent marker) and host (mouse) tissue cells in the mouse. In vivo research, but the mouse cannot be transplanted with human tissues or cells and survive due to its sound immune function, so it cannot be used for in vivo fluorescent tracer experiments on human tissue cells

Method used

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  • Method for establishing green fluorescence BALB/c nude mouse model and model application
  • Method for establishing green fluorescence BALB/c nude mouse model and model application
  • Method for establishing green fluorescence BALB/c nude mouse model and model application

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Experimental program
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Effect test

Embodiment 1

[0029] The establishment of embodiment 1 green fluorescent BALB / c nude mouse model

[0030] 1. Main materials and equipment:

[0031] BALB / c nude mice and green fluorescent protein transgenic mice C57BL / 6-Tg (CAG-EGFP) were purchased from the Model Animal Research Institute of Nanjing University (SCXK Su 2010-0001); independent ventilation cages were purchased from Suzhou Feng’s Experimental Animal Equipment Co., Ltd. (IVC-Ⅱ); the biological safety cabinet was purchased from Singapore Yisi Clean Equipment Co., Ltd. (ESCO); the small animal multi-mode live imaging instrument (In-vivo imaging system FX Pro) was purchased from Kodak Company. The fluorescent flashlight was purchased from Nightsea Company, PCR instrument (AB Company), flow cytometer (BD Company), fluorescent inverted microscope (Nikon Company), and CO2 incubator (Germany Heraeus Company). SLY series animal brain slice molds were purchased from Beijing Shuolinyuan Technology Co., Ltd. The frozen microtome was purc...

Embodiment 2

[0038] Embodiment 2 green fluorescent BALB / c nude mouse result and judgment

[0039] 1. Characteristics of green fluorescent BALB / c nude mice

[0040] The weight of adult mice is about 22-30g, and the litter size is 6-13 per generation. Homozygous (nu / nu, hairless) and heterozygous (nu / +, hairy) each accounted for about 50%. Such as figure 1 Under the excitation of 470nm blue light in the multi-mode small animal imaging system, the skin of nu / nu is green, the fur of nu / + is white, and the pinna and paws are green; gross anatomy, except hair (not including hair follicles) Red blood cells and peripheral blood do not emit green light, while other tissues and organs emit green light; tissue sections are excited by blue fluorescence microscope (CarlZeissAXIO Observer. Green fluorescence, but there are some differences in intensity.

[0041] exist figure 1 Middle 1 and 2 are hairless and hairy mice respectively, and No. 2 mouse does not emit green fluorescence because the hair ...

Embodiment 3

[0048] Embodiment 3 skin graft experiment

[0049] Flow cytometry analysis showed that the peripheral blood lymphocytes of BALB / c nude mouse inbred strain were MHC Class I type, and about 94% of peripheral blood lymphocytes were type d, which was consistent with the phenotype of BALB / c mice. From the 10th generation, 9 hairy 6-week-old mice (6-week-old female hairy mice) were anesthetized by intraperitoneal injection of 200 mg / kg of 10% chloral hydrate, and the skin with a diameter of 0.8 cm was taken from the back of the mouse, and the inner surface was trimmed. Subcutaneous tissue to dermis. It was exchanged with a skin graft of the same size taken from another mouse, sutured intermittently, the skin graft was bandaged under pressure, and the growth status of the skin graft was observed. The growth status of the transplanted skin: On the 10th day after the operation, the skin grafts were in good alignment, and there was no subcutaneous fluid. After 30 days, the growth of mi...

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Abstract

The invention discloses a method for establishing a green fluorescence BALB / c nude mouse model. The method includes: selecting female C57BL / 6-Tg (CAG-EGFP) mice and male Foxn1nu mice to perform hybrid to acquire F1 child generation hairy fluorescence mice, selecting the female mice, selecting the female mice having sexual maturation and the parental generation male Foxn1nu mice to perform mating (backcross) to acquire an F2 child generation; generating segregation of characters of the F2 child generation, selecting female hairy fluorescence mice and the parental generation male Foxn1nu mice to perform backcross, and completing a backcross cycle; performing the backcross cycle for ten times to acquire an F10 child generation, and establishing the green fluorescence BALB / c nude mouse model. The invention also discloses application of an animal model established by the method to a brain tumor transplanting model. The method uses a homogenous introduction method to establish the new strain model mice which can stably express EGFP, and can lay a firm foundation for human glioma orthotopic transplantation tracking and research on glioma and a micro environment thereof.

Description

technical field [0001] The invention belongs to the field of animal models used in the medical field, in particular to mouse models. Background technique [0002] Glioma is the most common primary refractory brain tumor in the brain. At present, domestic and foreign laboratories are conducting extensive basic research on glioma, including the mechanism of glioma, biological behavior, gene pathways and Biological therapy and gene therapy, etc., have improved the curative effect and improved prognosis to a certain extent. As a platform for basic research, it is crucial to establish an orthotopic transplantation model of human glioma. Although the research on such animal models is relatively mature, there are certain defects: the tumor model established with ordinary BALB / c nude mice cannot track and distinguish the tumor and the tumor microenvironment cells derived from the host; c The fluorescent nude mice crossed with fluorescent mice with normal immune function, because o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12Q1/68
CPCA01K67/0275C12Q1/686C12Q2565/125
Inventor 董军陈延明盛敏峰薛彦平刘加驰黄强
Owner THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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