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
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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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