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Method for establishing myasthenia gravis EAMG mouse skeletal muscle model

A method of establishing a mouse model, which is applied to radiological diagnostic equipment, image data processing, and diagnosis, and can solve the problems of strong subjectivity of clinical muscle strength indicators, insufficient evaluation of specific muscle groups, and lack of continuous observation indicators, etc. problems, to achieve the effect of shortening the molding time, reducing time consumption, and controlling costs

Active Publication Date: 2022-01-11
天津医科大学总医院空港医院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of this, the present invention aims to propose a method for establishing a skeletal muscle model of myasthenia gravis EAMG mice to solve the problem of strong subjectivity of clinical muscle strength indicators, insufficient evaluation of specific muscle groups, and lack of effective continuous observation indicators The problem

Method used

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  • Method for establishing myasthenia gravis EAMG mouse skeletal muscle model
  • Method for establishing myasthenia gravis EAMG mouse skeletal muscle model
  • Method for establishing myasthenia gravis EAMG mouse skeletal muscle model

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Preparation of DC cells with high antigen-presenting ability and verification of antigen-presenting ability

[0087] (1) Preparation of thymoma cells with high autophagy level:

[0088] a. Item preparation: plasmid vector pc3.1(+), human acute monocytic leukemia cell THP-1 (National Experimental Cell Resource Sharing Service Platform), fetal bovine serum (Clark Bio Company Cat#FB25015), Streptomyces lividans Vegetable mixed solution (Cat#10378016, Sigma-Aldrich, USA), β-sulfhydryl reducing agent (Cat#M8211, China Suo Laibao Biotechnology Co., Ltd.), 1640 basal medium (Gibco, USA, Cat#12633012), DMEM high glucose Basal medium (US Gibco Cat#11965092);

[0089] b. Thymoma cell line Thy0517 CGMCCNo.9328 (Thy0517 for short) preserved in liquid nitrogen was revived in a constant temperature water bath at 37°C, and the cells were transferred to the DMEM complete medium in the cell culture flask for culture, and the culture flask was placed in 37 Cultivate in a cell constant ...

Embodiment 2

[0109] Establishment of Immunoenhanced Myasthenia Gravis (EAMG) Mouse Model

[0110] a. Preparation of C57BL / 6 mice: Prepare 30 SPF-grade 6-8-week-old female experimental mice (C57BL / 6) (provided by Beijing Fukang Biotechnology Co., Ltd., license number: SCXK (Beijing) 2020- 0004);

[0111] b. Experimental animal feeding: The handling of animals during the experiment follows the relevant provisions of the "Regulations on the Administration of Experimental Animals" and complies with animal ethics management standards. The mice were kept in an SPF-grade animal room. The animal laboratory strictly followed the aseptic operation. Before entering the laboratory, the air laminar flow switch was turned on, and masks, hats, isolation gowns, and gloves were worn, and the animals entered after ten minutes of ventilation. After entering, turn on the ultraviolet lamp of the sterile workbench and irradiate it for more than 30 minutes before use. Before use, wipe with alcohol and ventilate...

Embodiment 3

[0121] Mouse Lennon Score and Existing System Validation

[0122] 1. Lennon score of EAMG mice

[0123] a. The score of clinical symptoms of mice is based on the Lennon scoring method, and the specific content is as follows:

[0124] Observe the activity of the mouse, if the activity is vigorous and the muscles are strong, then carry out the test. Put the mouse on the top grid of the cage, suspend the bottom of the tail of the mouse, and drag it repeatedly 20 times to stimulate the movement of the mouse, and then allow them to crawl freely. By observing whether the mouse has tremor, hunched posture, Muscle strength and signs of fatigue, thereby assessing their muscle weakness:

[0125] Grade 0, normal muscle strength, no muscle weakness even after exercise;

[0126] Grade 1, no clinical signs before the test, but weakness due to fatigue and decreased mobility after exercise;

[0127] Grade 2, clinical signs before test, hunched over, weak grip, chin on floor, inability to ...

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Abstract

The invention provides a method for establishing a myasthenia gravis EAMG mouse skeletal muscle model. According to the method, on the basis of artificially synthesizing an AChR peptide fragment immune induced C57BL / 6 mouse, an immune-enhanced EAMG mouse model is established in combination with infusion of in-vitro cultured dendritic cells with high antigen presentation capability, and meanwhile, the model is subjected to brand-new multi-model skeletal muscle detection. The invention discloses an immune-enhanced myasthenia gravis mouse model and establishes a novel skeletal muscle evaluation method. According to the method, quantitative analysis, functional image evaluation and tissue target determination on the affected skeletal muscle are realized. According to the method for establishing the myasthenia gravis EAMG mouse skeletal muscle model, the modeling time of the EAMG mouse is shortened, the modeling rate is increased, development of a myasthenia gravis animal experiment is facilitated, time consumed by the myasthenia gravis animal experiment is shortened, and the cost of the myasthenia gravis animal experiment is controlled.

Description

technical field [0001] The invention belongs to the field of myasthenia gravis, and in particular relates to a method for establishing a skeletal muscle model of myasthenia gravis EAMG mice. Background technique [0002] Myasthenia gravis (MG) is an autoimmune disease characterized by skeletal muscle weakness. The disease involves cellular immunity-mediated autoantibodies that lead to the involvement of acetylcholine receptors at the neuromuscular junction. Its prevalence rate is 150-300 people / 1 million people, and the prevalence rate of anti-acetylcholine receptor-positive myasthenia gravis (AChR MG) is 70-163 / 1 million people. Myasthenia gravis remains an intractable disease. [0003] Myasthenia gravis occurs alone or concurrently with other autoimmune diseases or tumor-related muscle diseases. Although the accurate immunophenotyping of myasthenia has been applied clinically, the target typing and intervention of different types of myasthenia are still the focus of rel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027G16B15/30G06T7/64G06T7/00A61B6/03A61B6/00G01N33/68
CPCA01K67/027G01N33/6854G16B15/30G06T7/0012G06T7/64A61B6/508A61B6/505A61B6/032A61B6/037A61B6/5217A01K2207/12A01K2227/105A01K2267/0375A01K2207/10A01K2267/0325G06T2207/10081G06T2207/30012
Inventor 张鹏路超王建垚张泽洋宋世辉
Owner 天津医科大学总医院空港医院
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