A kind of establishment method of EAMG mouse skeletal muscle model of myasthenia gravis

A method of establishing a mouse model, which is applied to radiological diagnostic instruments, analysis of two-dimensional or three-dimensional molecular structures, image enhancement, etc., can solve the problem of strong subjectivity of clinical muscle strength indicators and insufficient evaluation of specific muscle groups , Lack of continuous observation indicators and other issues, to achieve the effect of shortening the molding time, reducing time-consuming, and controlling costs

Active Publication Date: 2022-03-11
天津医科大学总医院空港医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A kind of establishment method of EAMG mouse skeletal muscle model of myasthenia gravis
  • A kind of establishment method of EAMG mouse skeletal muscle model of myasthenia gravis
  • A kind of establishment method of EAMG mouse skeletal muscle model of myasthenia gravis

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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 their 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 (Cat#FB25015, Clark Biological Company, USA), Streptomyces penicillin Sulphur mixture (Cat#10378016, Sigma-Aldrich, USA), β-thiol reducing agent (Cat#M8211, Soleibo Biotechnology Co., Ltd., China), 1640 basal medium (Gibco, USA, Cat#12633012), DMEM high glucose Basal medium (Gibco Cat#11965092, USA);

[0089] b. Resuscitate the thymoma cell line Thy0517 CGMCCNo.9328 (referred to as Thy0517) stored in liquid nitrogen in a 37°C constant temperature water bath, and transfer the cells to the DMEM complete medium in the cell culture flask for culture, and put the culture flask into 37 Cultivated in a constant tempera...

Embodiment 2

[0109] Establishment of immune-enhanced myasthenia gravis (EAMG) mouse model

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

[0111] b. Breeding of experimental animals: The disposal of animals in the experimental process is in accordance with the relevant provisions of the "Administrative Regulations on Laboratory Animals" and in line with animal ethics management standards. Mice are kept in SPF animal rooms. The animal laboratory strictly follows aseptic operation. Before entering the laboratory, turn on the air laminar flow switch, wear masks, hats, isolation gowns, and gloves, and enter after ten minutes of ventilation. After entering, turn on the UV lamp on the sterile workbench and irradiate it for more than 30 minutes before use. Wipe it with alcohol before use, and let it air dry.

[0112] c. Grouping of experimenta...

Embodiment 3

[0121] Mouse Lennon Score and Validation of Existing System

[0122] 1. Lennon score of EAMG mice

[0123] a. The clinical symptom score of mice is based on the Lennon scoring method, and the specific contents are:

[0124] Observe the activity of the mice. If the activity is vigorous and the muscles are strong, the test is carried out. Stimulate the mice to move by placing the mice on the top grid of the cage, levitating the bottom of the mice's tails and dragging them 20 times, and then allowing them to crawl freely by observing the mice for tremors, hunched posture, Muscle strength and signs of fatigue, thus 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 appeared before the test, hunched over, weak grip, chin on the floor, unable to raise head;

[0128...

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Abstract

The present invention provides a method for establishing a skeletal muscle model of EAMG mice with myasthenia gravis. The present invention is based on artificially synthesized AChR peptides to immunize and induce C57BL / 6 mice, combined with infusion of high antigen presentation ability cultivated in vitro Dendritic cells from dendritic cells, establish an immunoenhanced EAMG mouse model, and conduct a new multi-modal skeletal muscle detection on the model. method, the method realizes quantitative analysis, functional imaging evaluation, and tissue target determination of the involved skeletal muscle. The invention creates a method for establishing a skeletal muscle model of EAMG mice with myasthenia gravis, shortens the modeling time of EAMG mice and improves the modeling rate, is conducive to the development of animal experiments on myasthenia gravis, and is beneficial to reduce the risk of myasthenia gravis. Asthenia gravis animal experiments are time-consuming, which is beneficial to control the cost of myasthenia gravis animal experiments.

Description

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

Claims

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Application Information

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Patent Type & Authority Patents(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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