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

Active Publication Date: 2019-12-17
WENZHOU PEOPLES HOSPITAL
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Its side effects are mainly pulmonary toxicity, thyroid toxicity, cardiotoxicity, digestive system toxicity, etc. Among them, pulmonary toxicity is the most serious, with an incidence rate of 1%-10%. Pulmonary toxicity is mainly manifested as pulmonary fibrosis. Clinical lack of effective treatment measures, its mortality rate is as high as 33%, seriously affecting people's life and health
However, amiodarone has a definite and stable antiarrhythmic effect, and there is no other similar drug to replace it, and it is still widely used in clinical practice.

Method used

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  • A kind of culture method and detection method of rat fibrotic lung cells
  • A kind of culture method and detection method of rat fibrotic lung cells
  • A kind of culture method and detection method of rat fibrotic lung cells

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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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Abstract

The invention discloses a culture method and detection method for rat fibrotic lung cells. The culture method and detection method for rat fibrotic lung cells comprise the following steps: preparing a culture solution, a perfusion fluid and a rinsing solution; then sampling rat type II alveolar epithelial cells and carrying out primary culture; stimulating the type II alveolar epithelial cells with amiodarone so as to obtain fibrotic lung cells; and detecting the fibrotic lung cells. The culture method provided by the invention can acquire a great number of cells in a short period of time and accurately reflect the condition of amiodarone-induced pulmonary fibrosis in vivo and is of great significance to deep research on the influence of regulation and mechanism of the phenotypic transformation of the type II alveolar epithelial cells to delaying, or even reversion, of amiodarone-induced pulmonary fibrosis, and provides a research object for further exploration of the pathogenesis of amiodarone-induced pulmonary fibrosis at cell and molecule levels.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a method for culturing rat fibrotic lung cells and a detection method thereof. Background technique [0002] Amiodarone, also known as amiodarone, is an iodine-containing phenylpropanfuran derivative that is widely used in clinical practice as a class III antiarrhythmic drug, mainly for the treatment of ventricular arrhythmias and atrial fibrillation. Its side effects are mainly pulmonary toxicity, thyroid toxicity, cardiotoxicity, digestive system toxicity, etc. Among them, pulmonary toxicity is the most serious, with an incidence rate of 1%-10%. Pulmonary toxicity is mainly manifested as pulmonary fibrosis. Clinical lack of effective treatment measures, its mortality rate is as high as 33%, seriously affecting people's life and health. However, amiodarone has a definite and stable antiarrhythmic effect, and there is no other similar drug to replace it, and it is still widely used clini...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071G01N33/68
CPCC12N5/0688G01N33/6884G01N2800/12
Inventor 陈皓姜文兵王毅朱宁陈晓曙吴悠扬林伟李林锦胡逸人
Owner WENZHOU PEOPLES HOSPITAL
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