Effect of exosome in thoracic aortic dissection/aneurysm
A thoracic aorta and exosome technology, applied in the field of biomedicine, can solve the problems of high mortality, high recurrence rate, and many postoperative complications
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Embodiment 1
[0043] Example 1 The acquisition of exosomes
[0044] ① Uniformly inoculate UMSC cells in a logarithmic growth phase and in a good growth state in a 10 cm dish, centrifuge fetal bovine serum at a speed of 110,000 g for 7 hours to remove exosomes previously present in the serum, and use 10% exosome-free α-MEM culture medium of fetal bovine serum. The culture was continued for 48 hours, and when the cell density reached 90%, the old medium supernatant was collected, and the cells were subcultured.
[0045] ② Centrifuge the supernatant collected above at a speed of 2000g for 30min to remove dead cells and debris.
[0046] ③Use a straw to absorb the new supernatant, transfer it to a new 50mL centrifuge tube, add 1 / 2 volume of the supernatant PEG4000 exosome isolation reagent, shake it up and down, mix well, and extract at 4°C overnight.
[0047] ④After 24 hours, set the temperature of the centrifuge to 4°C, the speed to 13000g, and the time for 1 hour. After the centrifugation, ...
Embodiment 2
[0050] Embodiment 2 Construction of mouse thoracic aortic dissection / aneurysm model
[0051] Treat 3-week-old, C57BL / 6J male mice with β-aminopropionitrile (BAPN) drinking water, weigh BAPN powder, dissolve it in sterile double-distilled water, and the concentration of BAPN in the first and second weeks is 6mg / mL , the concentration of BAPN increased to 8mg / mL in the third and fourth weeks.
Embodiment 3
[0052] Embodiment 3 ultrasonic measurement diameter
[0053] Mice were anesthetized by respiratory inhalation with 2% isoflurane gas. After successful anesthesia, the skin was prepared on the chest and abdomen of the mice, and the hair on the body surface was removed to reduce the impact on the ultrasound images. The mice were placed in the supine position on the echocardiography platform, the limbs were fixed with adhesive tape on the flat-panel electrocardiogram monitoring position, the ultrasonic coupling agent was applied to the extremities, and the heated ultrasonic coupling agent was evenly applied to the chest and abdomen of the mouse. Using the MS400C ultrasonic probe, fix the ultrasonic probe above the echocardiography platform and adjust it down to a suitable position to avoid exerting too much pressure on the thoracic and abdominal cavity of the mouse while maintaining a clear image in the thoracic and abdominal cavity. Find the long-axis section of the heart along...
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