A kind of culture method and detection method of rat fibrotic lung cells
A technology of detection method and cultivation method, applied in the medical field, can solve problems such as unknown pathogenesis, lack of effective treatment measures, and affecting people's life and health, and achieve the effect of increasing the incidence rate
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Embodiment 1
[0019] Step 1. Preparation of culture medium, perfusion solution and cleaning solution: prepare DMEM medium, then add fetal bovine serum, 100U / ml penicillin and 100U / ml streptomycin at a final concentration of 100ml / L; prepare perfusion solution, and use PBS to prepare Trypsin with a mass fraction of 0.1%, and then add penicillin at a final concentration of 100U / ml, streptomycin at a concentration of 100U / ml and EDTA with a mass fraction of 0.02%. ml penicillin and 100U / ml streptomycin;
[0020] Step 2. Material collection: Rats were anesthetized by intraperitoneal injection of 3% pentobarbital 0.2ml / 100g, intraperitoneal injection of heparin sodium 2800U, the rats were sacrificed, and the chest cavity was opened, and the cleaning solution was passed through the right ventricle pulmonary artery Wash away the blood, then take out the lungs, perfuse the alveolar cavity with perfusate, incubate in a vibrating water bath at 37°C for 10 minutes, replace with new perfusate, and incu...
Embodiment 2
[0025] Step 1. Preparation of culture medium, perfusion solution and cleaning solution: prepare DMEM medium, then add fetal bovine serum, 100U / ml penicillin and 100U / ml streptomycin at a final concentration of 100ml / L; prepare perfusion solution, and use PBS to prepare Trypsin with a mass fraction of 0.1%, and then add penicillin at a final concentration of 100U / ml, streptomycin at a concentration of 100U / ml and EDTA with a mass fraction of 0.02%. ml penicillin and 100U / ml streptomycin;
[0026] Step 2, collecting materials: Rats were anesthetized by intraperitoneal injection of 3% pentobarbital 0.3ml / 100g, intraperitoneal injection of heparin sodium 3000U, the rats were sacrificed and the chest cavity was opened, and the cleaning solution was passed through the right ventricle pulmonary artery Wash off the blood, then take out the lungs, perfuse the alveolar cavity with perfusion fluid, incubate in a vibrating water bath at 37°C for 10 min, replace with new perfusate, incubat...
Embodiment 3
[0031] Step 1. Preparation of culture medium, perfusion solution and cleaning solution: prepare DMEM medium, then add fetal bovine serum, 100U / ml penicillin and 100U / ml streptomycin at a final concentration of 100ml / L; prepare perfusion solution, and use PBS to prepare Trypsin with a mass fraction of 0.1%, and then add penicillin at a final concentration of 100U / ml, streptomycin at a concentration of 100U / ml and EDTA with a mass fraction of 0.02%. ml penicillin and 100U / ml streptomycin;
[0032] Step 2, collecting materials: Rats were anesthetized by intraperitoneal injection of 3% pentobarbital 0.3ml / 100g, intraperitoneal injection of heparin sodium 3000U, the rats were sacrificed and the chest cavity was opened, and the cleaning solution was passed through the right ventricle pulmonary artery Wash away the blood, then remove the lungs, perfuse the alveolar space with perfusion fluid, incubate in a shaking water bath at 37°C for 30 minutes, remove the trachea, large airways a...
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